[ { "id": "https://authors.library.caltech.edu/records/ndqwz-kx867", "eprint_status": "archive", "datestamp": "2023-10-03 17:40:47", "lastmod": "2024-01-09 22:20:44", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Blumenfeld-Zack", "name": { "family": "Blumenfeld", "given": "Zack" }, "orcid": "0000-0002-4627-5582" }, { "id": "Bera-Kallol", "name": { "family": "Bera", "given": "Kallol" } }, { "id": "Castr\u00e9n-Eero", "name": { "family": "Castr\u00e9n", "given": "Eero" }, "orcid": "0000-0002-1402-2791" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Antidepressants enter cells, organelles, and membranes", "ispublished": "pub", "full_text_status": "public", "keywords": "Psychiatry and Mental health; Pharmacology", "note": "
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
\n\nThe group of HAL has been funded by the National Institute of Mental Health (MH1230823), the National Institute of General Medical Science (GM123582), National Institute on Drug Abuse (DA043829, DA049140), and the California Tobacco-Related Disease Research Program (27IP-0057). KB was supported by the Della Martin Foundation and the Howard Hughes Medical Institute. The group of EC was supported by the Academy of Finland (294710, 303124, 307416 and 327192), Sigrid Jus\u00e9lius Foundation, and Jane and Aatos Erkko Foundation.
\n\nAll authors wrote and edited all portions of the manuscript. KB provided final formatting of Table 1 and the Supplementary Files and also conceived the Figures.
\n\nThe authors declare no competing interests.
", "abstract": "We begin by summarizing several examples of antidepressants whose therapeutic actions begin when they encounter their targets in the cytoplasm or in the lumen of an organelle. These actions contrast with the prevailing view that most neuropharmacological actions begin when drugs engage their therapeutic targets at extracellular binding sites of plasma membrane targets\u2014ion channels, receptors, and transporters. We review the chemical, pharmacokinetic, and pharmacodynamic principles underlying the movements of drugs into subcellular compartments. We note the relationship between protonation-deprotonation events and membrane permeation of antidepressant drugs. The key properties relate to charge and hydrophobicity/lipid solubility, summarized by the parameters LogP, pK\u2090, and LogD(pH7.4). The classical metric, volume of distribution (Vd), is unusually large for some antidepressants and has both supracellular and subcellular components. A table gathers structures, LogP, PK\u2090, LogD(pH7.4), and V_d data and/or calculations for most antidepressants and antidepressant candidates. The subcellular components, which can now be measured in some cases, are dominated by membrane binding and by trapping in the lumen of acidic organelles. For common antidepressants, such as selective serotonin reuptake inhibitors (SSRIs) and serotonin/norepinephrine reuptake inhibitors (SNRIs), the target is assumed to be the eponymous reuptake transporter(s), although in fact the compartment of target engagement is unknown. We review special aspects of the pharmacokinetics of ketamine, ketamine metabolites, and other rapidly acting antidepressants (RAADs) including methoxetamine and scopolamine, psychedelics, and neurosteroids. Therefore, the reader can assess properties that markedly affect a drug's ability to enter or cross membranes\u2014and therefore, to interact with target sites that face the cytoplasm, the lumen of organelles, or a membrane. In the current literature, mechanisms involving intracellular targets are termed \"location-biased actions\" or \"inside-out pharmacology\". Hopefully, these general terms will eventually acquire additional mechanistic details.
", "date": "2023-10-02", "date_type": "published", "publication": "Neuropsychopharmacology", "publisher": "Nature Publishing Group", "issn": "0893-133X", "official_url": "https://authors.library.caltech.edu/records/ndqwz-kx867", "funders": { "items": [ { "grant_number": "MH1230823" }, { "grant_number": "GM123582" }, { "grant_number": "DA043829" }, { "grant_number": "DA049140" }, { "agency": "Della Martin Foundation" }, {}, { "grant_number": "294710" }, { "grant_number": "303124" }, { "grant_number": "307416" }, { "grant_number": "327192" }, {}, {} ] }, "local_group": { "items": [ { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1038/s41386-023-01725-x", "resource_type": "article", "pub_year": "2023", "author_list": "Blumenfeld, Zack; Bera, Kallol; et el." }, { "id": "https://authors.library.caltech.edu/records/frqfe-5rg44", "eprint_id": 121968, "eprint_status": "archive", "datestamp": "2023-08-22 20:50:01", "lastmod": "2023-12-22 23:38:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Walker-Noah-B", "name": { "family": "Walker", "given": "Noah B." }, "orcid": "0000-0002-8986-5611" }, { "id": "Yan-Yijin", "name": { "family": "Yan", "given": "Yijin" } }, { "id": "Tapia-Melissa-A", "name": { "family": "Tapia", "given": "Melissa A." } }, { "id": "Tucker-Brenton-R", "name": { "family": "Tucker", "given": "Brenton R." } }, { "id": "Thomas-Leanne-N", "name": { "family": "Thomas", "given": "Leanne N." } }, { "id": "George-Brianna-E", "name": { "family": "George", "given": "Brianna E." }, "orcid": "0000-0002-5234-0889" }, { "id": "West-Alyssa-M", "name": { "family": "West", "given": "Alyssa M." } }, { "id": "Marotta-Christopher-B", "name": { "family": "Marotta", "given": "Christopher B." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Holleran-Katherine-M", "name": { "family": "Holleran", "given": "Katherine M." } }, { "id": "Jones-Sara-R", "name": { "family": "Jones", "given": "Sara R." }, "orcid": "0000-0002-3424-7576" }, { "id": "Drenan-Ryan-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" } ] }, "title": "\u03b22 nAChR Activation on VTA DA Neurons Is Sufficient for Nicotine Reinforcement in Rats", "ispublished": "pub", "full_text_status": "public", "keywords": "General Medicine; General Neuroscience", "note": "\u00a9 2023 Walker et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. \n\nWe thank members of the Drenan and Jones lab for helpful discussion. \n\nThis work was supported by National Institutes of Health Grants DA040626 and DA035942 (to R.M.D.), DA048490 and DA006634 (to S.R.J.), DA049504 and NS117356 (to B.E.G.) and by The Regents of the University of California, Research Grants Program Office, Tobacco Related Disease Research Program T29IR0455 (to D.A.D.). \n\nThe authors declare no competing financial interests.\n\nPublished - ENEURO.0449-22.2023.full.pdf
", "abstract": "Mesolimbic nicotinic acetylcholine receptor (nAChRs) activation is necessary for nicotine reinforcement behavior, but it is unknown whether selective activation of nAChRs in the dopamine (DA) reward pathway is sufficient to support nicotine reinforcement. In this study, we tested the hypothesis that activation of \u03b22-containing (\u03b22*) nAChRs on VTA neurons is sufficient for intravenous nicotine self-administration (SA). We expressed \u03b22 nAChR subunits with enhanced sensitivity to nicotine (referred to as \u03b22Leu9\u2032Ser) in the VTA of male Sprague Dawley (SD) rats, enabling very low concentrations of nicotine to selectively activate \u03b22* nAChRs on transduced neurons. Rats expressing \u03b22Leu9\u2032Ser subunits acquired nicotine SA at 1.5\u2009\u03bcg/kg/infusion, a dose too low to support acquisition in control rats. Saline substitution extinguished responding for 1.5\u2009\u03bcg/kg/inf, verifying that this dose was reinforcing. \u03b22Leu9\u2032Ser nAChRs also supported acquisition at the typical training dose in rats (30\u2009\u03bcg/kg/inf) and reducing the dose to 1.5\u2009\u03bcg/kg/inf caused a significant increase in the rate of nicotine SA. Viral expression of \u03b22Leu9\u2032Ser subunits only in VTA DA neurons (via TH-Cre rats) also enabled acquisition of nicotine SA at 1.5\u2009\u03bcg/kg/inf, and saline substitution significantly attenuated responding. Next, we examined electrically-evoked DA release in slices from \u03b22Leu9\u2032Ser rats with a history of nicotine SA. Single-pulse evoked DA release and DA uptake rate were reduced in \u03b22Leu9\u2032Ser NAc slices, but relative increases in DA following a train of stimuli were preserved. These results are the first to report that \u03b22* nAChR activation on VTA neurons is sufficient for nicotine reinforcement in rats.", "date": "2023-05", "date_type": "published", "publication": "eNeuro", "volume": "10", "number": "5", "publisher": "Society for Neuroscience", "pagerange": "Art. No. ENEURO.0449-22.2023", "id_number": "CaltechAUTHORS:20230622-889344000.3", "issn": "2373-2822", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20230622-889344000.3", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA040626" }, { "agency": "NIH", "grant_number": "DA035942" }, { "agency": "NIH", "grant_number": "DA048490" }, { "agency": "NIH", "grant_number": "DA006634" }, { "agency": "NIH", "grant_number": "DA049504" }, { "agency": "NIH", "grant_number": "NS117356" }, { "agency": "Regents of the University of California" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "T29IR0455" } ] }, "local_group": { "items": [ { "id": "Tianqiao-and-Chrissy-Chen-Institute-for-Neuroscience" }, { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1523/eneuro.0449-22.2023", "pmcid": "PMC10216253", "primary_object": { "basename": "ENEURO.0449-22.2023.full.pdf", "url": "https://authors.library.caltech.edu/records/frqfe-5rg44/files/ENEURO.0449-22.2023.full.pdf" }, "resource_type": "article", "pub_year": "2023", "author_list": "Walker, Noah B.; Yan, Yijin; et el." }, { "id": "https://authors.library.caltech.edu/records/dh9ft-yqb12", "eprint_id": 121383, "eprint_status": "archive", "datestamp": "2023-10-06 16:18:51", "lastmod": "2024-01-09 22:19:31", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nichols-Aaron-L", "name": { "family": "Nichols", "given": "Aaron L." } }, { "id": "Blumenfeld-Zachary", "name": { "family": "Blumenfeld", "given": "Zack" } }, { "id": "Luebbert-Laura", "name": { "family": "Luebbert", "given": "Laura" } }, { "id": "Knox-Hailey-J", "name": { "family": "Knox", "given": "Hailey J." } }, { "id": "Muthusamy-Anand-K", "name": { "family": "Muthusamy", "given": "Anand K." } }, { "id": "Marvin-Jonathan-S", "name": { "family": "Marvin", "given": "Jonathan S." } }, { "id": "Kim-Charlene-M", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "Grant-Stephen-N", "name": { "family": "Grant", "given": "Stephen N." } }, { "id": "Walton-David-P", "name": { "family": "Walton", "given": "David P." } }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Hammar-Rebekkah", "name": { "family": "Hammar", "given": "Rebekkah" } }, { "id": "Looger-Loren-L", "name": { "family": "Looger", "given": "Loren" } }, { "id": "Artursson-Per", "name": { "family": "Artursson", "given": "Per" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Selective Serotonin Reuptake Inhibitors within Cells: Temporal Resolution in Cytoplasm, Endoplasmic Reticulum, and Membrane", "ispublished": "pub", "full_text_status": "public", "keywords": "General Neuroscience", "note": "\u00a9 2023 the authors under SfN exclusive license. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license. \n\nA.L.N. was supported by California Tobacco-Related Disease Research Program (TRDRP) Grant 27FT-0022; H.A.L. was supported by California TRDRP Grant 27IP-0057, National Institutes of Health (NIH)\u2013National Institute of General Medical Sciences (NIGMS) Grant GM-123582, NIH-National Institute on Drug Abuse grant NS105595, and NIH\u2013National Institute of Mental Health Grant MH120823; D.A.D. was supported by California TRDRP Grant T29IR0455; A.K.M. was supported by NIH\u2013National Institute on Drug Abuse Grant DA049140 and NIH\u2013NIGMS Grant GM7616; J.S.M. and L. Looger were supported by the Howard Hughes Medical Institute; L. Luebbert was supported by Leiden University International Studies Fund Grant LISF L18020-1-45; P.A. was supported by Swedish Research Council Grant 01586; and R.H. was supported by the European Union Horizon 2020 Research and Innovation Program under Marie Sk\u0142odowska-Curie Grant Agreement 956851. We thank Stefan Petrovic for stewardship of the isothermal titration calorimeter in the Caltech Center for Molecular Medicine; the Gradinaru lab and Caltech CLOVER Center for help with viral vectors; Andres Collazo and Giada Spigolon at the Caltech Biological Imaging Facility; Stefan Trapp for furnishing and explaining the original version of the Microsoft Excel workbook for acid trapping; Zoe Beatty, Kallol Bera, Eve Fine, Shan Huang, Elaine Lin, Stephen Mayo, Lin Tian, Andrea Treyer, Elizabeth Unger, and Lu Wei for advice and guidance; and Purnima Deshpande for lab management. \n\nAuthor Contributions. Author contributions: A.L.N., Z.B., L. Luebbert, H.J.K., A.K.M., J.S.M., C.H.K., S.N.G., D.P.W., R.H., L. Looger, P.A., D.A.D., and H.A.L. designed research; A.L.N., Z.B., L. Luebbert, H.J.K., A.K.M., J.S.M., C.H.K., S.N.G., D.P.W., and R.H. performed research; A.L.N., Z.B., L. Luebbert, H.J.K., A.K.M., J.S.M., S.N.G., D.P.W., B.N.C., R.H., P.A., D.A.D., and H.A.L. analyzed data; A.L.N., Z.B., L. Looger, and H.A.L. wrote the paper. \n\nA.L.N. and Z.B. share co-first authorship.\n\nThe authors declare no competing financial interests.", "abstract": "Selective serotonin reuptake inhibitors (SSRIs) are the most prescribed treatment for individuals experiencing major depressive disorder. The therapeutic mechanisms that take place before, during, or after SSRIs bind the serotonin transporter (SERT) are poorly understood, partially because no studies exist on the cellular and subcellular pharmacokinetic properties of SSRIs in living cells. We studied escitalopram and fluoxetine using new intensity-based, drug-sensing fluorescent reporters targeted to the plasma membrane, cytoplasm, or endoplasmic reticulum (ER) of cultured neurons and mammalian cell lines. We also used chemical detection of drug within cells and phospholipid membranes. The drugs attain equilibrium in neuronal cytoplasm and ER at approximately the same concentration as the externally applied solution, with time constants of a few s (escitalopram) or 200\u2013300 s (fluoxetine). Simultaneously, the drugs accumulate within lipid membranes by \u226518-fold (escitalopram) or 180-fold (fluoxetine), and possibly by much larger factors. Both drugs leave cytoplasm, lumen, and membranes just as quickly during washout. We synthesized membrane-impermeant quaternary amine derivatives of the two SSRIs. The quaternary derivatives are substantially excluded from membrane, cytoplasm, and ER for >2.4 h. They inhibit SERT transport-associated currents sixfold or 11-fold less potently than the SSRIs (escitalopram or fluoxetine derivative, respectively), providing useful probes for distinguishing compartmentalized SSRI effects. Although our measurements are orders of magnitude faster than the therapeutic lag of SSRIs, these data suggest that SSRI\u2013SERT interactions within organelles or membranes may play roles during either the therapeutic effects or the antidepressant discontinuation syndrome.\n\nSIGNIFICANCE STATEMENT:\nSelective serotonin reuptake inhibitors stabilize mood in several disorders. In general, these drugs bind to SERT, which clears serotonin from CNS and peripheral tissues. SERT ligands are effective and relatively safe; primary care practitioners often prescribe them. However, they have several side effects and require 2\u20136 weeks of continuous administration until they act effectively. How they work remains perplexing, contrasting with earlier assumptions that the therapeutic mechanism involves SERT inhibition followed by increased extracellular serotonin levels. This study establishes that two SERT ligands, fluoxetine and escitalopram, enter neurons within minutes, while simultaneously accumulating in many membranes. Such knowledge will motivate future research, hopefully revealing where and how SERT ligands engage their therapeutic target(s).", "date": "2023-03-29", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "43", "number": "13", "publisher": "Society for Neuroscience", "pagerange": "2222-2241", "id_number": "CaltechAUTHORS:20230512-807738000.3", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20230512-807738000.3", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "27FT-0022" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "27IP-0057" }, { "agency": "NIH", "grant_number": "GM-123582" }, { "agency": "NIH", "grant_number": "NS105595" }, { "agency": "NIH", "grant_number": "MH120823" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "T29IR0455" }, { "agency": "NIH", "grant_number": "DA049140" }, { "agency": "NIH", "grant_number": "GM7616" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "Leiden University", "grant_number": "LISF L18020-1-45" }, { "agency": "Swedish Research Council", "grant_number": "01586" }, { "agency": "Marie Curie Fellowship", "grant_number": "956851" } ] }, "local_group": { "items": [ { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1523/jneurosci.1519-22.2022", "pmcid": "PMC10072302", "primary_object": { "basename": "zns2222.pdf", "url": "https://authors.library.caltech.edu/records/dh9ft-yqb12/files/zns2222.pdf" }, "resource_type": "article", "pub_year": "2023", "author_list": "Nichols, Aaron L.; Blumenfeld, Zack; et el." }, { "id": "https://authors.library.caltech.edu/records/kvgc3-j1k69", "eprint_id": 116865, "eprint_status": "archive", "datestamp": "2023-08-22 17:39:02", "lastmod": "2023-12-22 23:15:38", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Knox-Hailey-J", "name": { "family": "Knox", "given": "Hailey J." }, "orcid": "0000-0003-0608-2855" }, { "id": "Rego-Campello-H", "name": { "family": "Rego Campello", "given": "Hugo" }, "orcid": "0000-0001-8588-0198" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Gallagher-Timothy", "name": { "family": "Gallagher", "given": "Timothy" }, "orcid": "0000-0002-3544-327X" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Characterization of Binding Site Interactions and Selectivity Principles in the \u03b13\u03b24 Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "public", "keywords": "Colloid and Surface Chemistry; Biochemistry; General Chemistry; Catalysis", "note": "We would like to thank Jonathan Wang and Purnima Deshpande for harvesting and preparing oocytes from Xenopus laevis. We thank Stephen Grant, Maria Constanza Maldifassi Gatica, and Christopher Marotta for useful discussions and suggestions. We thank Annet Blom for guidance on initial expression and stoichiometry determination for \u03b13\u03b24. We also acknowledge Achieve Life Sciences for their generous gift of (\u2212)-cytisine. This work was supported by in part by funds provided by The Regents of the University of California, Research Grants Program Office, Tobacco Related Disease Research Program (Grant No. T29IR0455 to D.A.D.). The opinions, findings, and conclusions herein are those of the authors and do not necessarily represent those of The Regents of the University of California, or any of its programs.", "abstract": "Nicotinic acetylcholine receptors (nAChRs) play an important role in neurotransmission and are also involved in addiction and several disease states. There is significant interest in therapeutic targeting of nAChRs; however, achieving selectivity for one subtype over others has been a longstanding challenge, given the close structural similarities across the family. Here, we characterize binding interactions in the \u03b13\u03b24 nAChR subtype via structure\u2013function studies involving noncanonical amino acid mutagenesis and two-electrode voltage clamp electrophysiology. We establish comprehensive binding models for both the endogenous neurotransmitter ACh and the smoking cessation drug cytisine. We also use a panel of C(10)-substituted cytisine derivatives to probe the effects of subtle changes in the ligand structure on binding. By comparing our results to those obtained for the well-studied \u03b14\u03b22 subtype, we identify several features of both the receptor and agonist structure that can be utilized to enhance selectivity for either \u03b13\u03b24 or \u03b14\u03b22. Finally, we characterize binding interactions of the \u03b13\u03b24-selective partial agonist AT-1001 to determine factors that contribute to its selectivity. These results shed new light on the design of selective nAChR-targeted ligands and can be used to inform the design of improved therapies with minimized off-target effects.", "date": "2022-09-07", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "144", "number": "35", "publisher": "American Chemical Society", "pagerange": "16101-16117", "id_number": "CaltechAUTHORS:20220909-232673000", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220909-232673000", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "T29IR0455" } ] }, "local_group": { "items": [ { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1021/jacs.2c06495", "resource_type": "article", "pub_year": "2022", "author_list": "Knox, Hailey J.; Rego Campello, Hugo; et el." }, { "id": "https://authors.library.caltech.edu/records/djcpp-84e21", "eprint_id": 116720, "eprint_status": "archive", "datestamp": "2023-08-22 17:26:54", "lastmod": "2023-12-22 23:31:45", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Harrison-Neil-L", "name": { "family": "Harrison", "given": "Neil L." }, "orcid": "0000-0002-8013-7765" }, { "id": "Abbott-Geoffrey-W", "name": { "family": "Abbott", "given": "Geoffrey W." }, "orcid": "0000-0003-4552-496X" }, { "id": "Gentzsch-Martina", "name": { "family": "Gentzsch", "given": "Martina" }, "orcid": "0000-0002-9435-0321" }, { "id": "Aleksandrov-Andrei", "name": { "family": "Aleksandrov", "given": "Andrei" } }, { "id": "Moroni-Anna", "name": { "family": "Moroni", "given": "Anna" }, "orcid": "0000-0002-1860-406X" }, { "id": "Thiel-Gerhard", "name": { "family": "Thiel", "given": "Gerhard" } }, { "id": "Grant-Stephen", "name": { "family": "Grant", "given": "Stephen" }, "orcid": "0000-0003-0923-8886" }, { "id": "Nichols-Colin-G", "name": { "family": "Nichols", "given": "Colin G." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Hartel-Andreas", "name": { "family": "Hartel", "given": "Andreas" } }, { "id": "Shepard-Kenneth-L", "name": { "family": "Shepard", "given": "Kenneth" }, "orcid": "0000-0003-0665-6775" }, { "id": "Garcia-David-Cabrera", "name": { "family": "Garcia", "given": "David Cabrera" }, "orcid": "0000-0001-9609-9102" }, { "id": "Yazawa-Masayuki", "name": { "family": "Yazawa", "given": "Masayuki" } } ] }, "title": "How many SARS-CoV-2 \"viroporins\" are really ion channels?", "ispublished": "pub", "full_text_status": "public", "keywords": "General Agricultural and Biological Sciences; General Biochemistry, Genetics and Molecular Biology; Medicine (miscellaneous)", "abstract": "The SARS-CoV-2 virus uses a small number of viral proteins to enter host cells and disrupt their activity, including the spike (S), membrane (M), and envelope (E) proteins, as well as a number of accessory proteins of unknown function (Orf3a, Orf8, Orf10, etc.), some of which may function to alter ion flux across membranes. Toft-Bertelsen et al.1 recently reported that the drug amantadine may interact with virally encoded ion channels and proposed that inhibitors of these \"viroporins\" might have therapeutic use in COVID-19. We concur with the idea that the E protein of SARS-CoV-2 can form an ion channel and that amantadine inhibits this channel, but we suggest below that a number of additional specific criteria need to be met in order for SARS-CoV-2 accessory proteins (Orf3a, Orf8, Orf10) to be accepted as having ion channel activity, and that further work will be necessary. This field represents a neglected area of overlap between biophysics and virology that continues to be understudied and underfunded and\nclearly merits greater attention from the relevant funding agencies.", "date": "2022-08-25", "date_type": "published", "publication": "Communications Biology", "volume": "5", "number": "1", "publisher": "Nature Publishing Group", "id_number": "CaltechAUTHORS:20220906-252586000", "issn": "2399-3642", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220906-252586000", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM130377" } ] }, "local_group": { "items": [ { "id": "COVID-19" }, { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1038/s42003-022-03669-2", "resource_type": "article", "pub_year": "2022", "author_list": "Harrison, Neil L.; Abbott, Geoffrey W.; et el." }, { "id": "https://authors.library.caltech.edu/records/e0207-xyw78", "eprint_id": 113622, "eprint_status": "archive", "datestamp": "2023-08-20 07:43:36", "lastmod": "2023-12-22 23:15:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Muthusamy-Anand-K", "name": { "family": "Muthusamy", "given": "Anand K." }, "orcid": "0000-0003-1041-914X" }, { "id": "Kim-Charlene-M", "name": { "family": "Kim", "given": "Charlene H." }, "orcid": "0000-0003-4048-5244" }, { "id": "Virgil-Scott-C", "name": { "family": "Virgil", "given": "Scott C." }, "orcid": "0000-0001-8586-5641" }, { "id": "Knox-Hailey-J", "name": { "family": "Knox", "given": "Hailey J." }, "orcid": "0000-0003-0608-2855" }, { "id": "Marvin-Jonathan-S", "name": { "family": "Marvin", "given": "Jonathan S." }, "orcid": "0000-0003-2294-4515" }, { "id": "Nichols-Aaron-L", "name": { "family": "Nichols", "given": "Aaron L." }, "orcid": "0000-0001-9341-0049" }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce N." }, "orcid": "0000-0003-2913-6238" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Looger-Loren-L", "name": { "family": "Looger", "given": "Loren L." }, "orcid": "0000-0002-7531-1757" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Three Mutations Convert the Selectivity of a Protein Sensor from Nicotinic Agonists to S-Methadone for Use in Cells, Organelles, and Biofluids", "ispublished": "pub", "full_text_status": "public", "keywords": "Fluorescence, Genetics, Sensors, Pharmaceuticals, Biotechnology", "note": "\u00a9 2022 The Authors. Published by American Chemical Society.\nUnder an Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0). \n\nReceived: March 1, 2022; Published: April 21, 2022. \n\nThe Caltech Center for Catalysis and Chemical Synthesis supported by the Beckman Institute supported the chiral resolution work. Dr. Viviana Gradinaru and the CLOVER Center at Caltech provided plasmids and advice for AAV production. mApple-Golgi-7 was a gift from Dr. Michael Davidson (Addgene plasmid #54907). Andres Collazo and Giada Spigolon manage the Biological Imaging Facility supported by the Beckman Institute and advised on confocal imaging. The Proteome Exploration Laboratory was supported by NIH OD010788, NIH OD020013, the Betty and Gordon Moore Foundation through Grant GBMF775, and the Beckman Institute at Caltech. Dr. Wei Gao, Dr. You Yu, and Heather Lukas gathered human sweat. We thank Dr. Luke Lavis for contributive discussions. We thank Dr. Aiden Aceves and Dr. Stephen Mayo for advice on docking, Theodore Chin for assistance in cloning, and Purnima Deshpande for excellent lab management. \n\nThis work was supported by the National Institute on Drug Abuse (NIDA) Grant DA043829, National Institute of General Medical Sciences (NIGMS) Grant GM-123582, and Janelia Research Campus, HHMI. H.A.L. was supported by DA043829 and GM-123582. A.K.M. was supported by DA043829, NIGMS fellowship 5T32GM007616, and National Institute of Neurological Disorders and Stroke fellowship T32NS105595. D.A.D. and H.J.K. were supported by the Tobacco-Related Disease Research Program (TRDRP) Grant T29IR0455. A.L.N. was supported by TRDRP fellowship 27FT-0022. L.L.L. and J.S.M. were supported by Janelia Research Campus, HHMI. \n\nThe authors declare the following competing financial interest(s): Anand K. Muthusamy, Henry A. Lester, Loren L. Looger, and Jonathan S. Marvin have filed a patent application that includes iS-methadoneSnFR.\nConstructs reported in this manuscript will be deposited in Addgene. Sequences and dose response metrics will be made available in a GitHub repository.\n\nPublished - jacs.2c02323.pdf
Submitted - 2022.02.24.481226v2.full.pdf
Supplemental Material - ja2c02323_si_001.pdf
", "abstract": "We report a reagentless, intensity-based S-methadone fluorescent sensor, iS-methadoneSnFR, consisting of a circularly permuted GFP inserted within the sequence of a mutated bacterial periplasmic binding protein (PBP). We evolved a previously reported nicotine-binding PBP to become a selective S-methadone-binding sensor, via three mutations in the PBP's second shell and hinge regions. iS-methadoneSnFR displays the necessary sensitivity, kinetics, and selectivity\u2500notably enantioselectivity against R-methadone\u2500for biological applications. Robust iS-methadoneSnFR responses in human sweat and saliva and mouse serum enable diagnostic uses. Expression and imaging in mammalian cells demonstrate that S-methadone enters at least two organelles and undergoes acid trapping in the Golgi apparatus, where opioid receptors can signal. This work shows a straightforward strategy in adapting existing PBPs to serve real-time applications ranging from subcellular to personal pharmacokinetics.", "date": "2022-05-18", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "144", "number": "19", "publisher": "American Chemical Society", "pagerange": "8480-8486", "id_number": "CaltechAUTHORS:20220228-619163000", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220228-619163000", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA043829" }, { "agency": "NIH", "grant_number": "GM-123582" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5T32GM007616" }, { "agency": "NIH", "grant_number": "T32NS105595" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "T29IR0455" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "27FT-0022" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "Caltech Beckman Institute" }, { "agency": "NIH", "grant_number": "OD010788" }, { "agency": "NIH", "grant_number": "OD020013" }, { "agency": "Gordon and Betty Moore Foundation", "grant_number": "GBMF775" } ] }, "local_group": { "items": [ { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1021/jacs.2c02323", "pmcid": "PMC9121368", "primary_object": { "basename": "2022.02.24.481226v2.full.pdf", "url": "https://authors.library.caltech.edu/records/e0207-xyw78/files/2022.02.24.481226v2.full.pdf" }, "related_objects": [ { "basename": "ja2c02323_si_001.pdf", "url": "https://authors.library.caltech.edu/records/e0207-xyw78/files/ja2c02323_si_001.pdf" }, { "basename": "jacs.2c02323.pdf", "url": "https://authors.library.caltech.edu/records/e0207-xyw78/files/jacs.2c02323.pdf" } ], "resource_type": "article", "pub_year": "2022", "author_list": "Muthusamy, Anand K.; Kim, Charlene H.; et el." }, { "id": "https://authors.library.caltech.edu/records/ft222-e3n43", "eprint_id": 111334, "eprint_status": "archive", "datestamp": "2023-08-20 06:30:30", "lastmod": "2023-12-22 23:38:46", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nichols-Aaron-L", "name": { "family": "Nichols", "given": "Aaron L." }, "orcid": "0000-0001-9341-0049" }, { "id": "Blumenfeld-Zachary", "name": { "family": "Blumenfeld", "given": "Zack" }, "orcid": "0000-0002-4627-5582" }, { "id": "Fan-Chengcheng", "name": { "family": "Fan", "given": "Chengcheng" }, "orcid": "0000-0003-4213-5758" }, { "id": "Luebbert-Laura", "name": { "family": "Luebbert", "given": "Laura" }, "orcid": "0000-0003-1379-2927" }, { "id": "Blom-Annet-E-M", "name": { "family": "Blom", "given": "Annet E. M." }, "orcid": "0000-0002-7441-4893" }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Marvin-Jonathan-S", "name": { "family": "Marvin", "given": "Jonathan S." }, "orcid": "0000-0003-2294-4515" }, { "id": "Borden-Philip-M", "name": { "family": "Borden", "given": "Philip M." }, "orcid": "0000-0003-1653-7067" }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "Muthusamy-Anand-K", "name": { "family": "Muthusamy", "given": "Anand K." }, "orcid": "0000-0003-1041-914X" }, { "id": "Shivange-Amol-V", "name": { "family": "Shivange", "given": "Amol V." }, "orcid": "0000-0002-4169-2969" }, { "id": "Knox-Hailey-J", "name": { "family": "Knox", "given": "Hailey J." }, "orcid": "0000-0003-0608-2855" }, { "id": "Rego-Campello-Hugo", "name": { "family": "Rego Campello", "given": "Hugo" }, "orcid": "0000-0001-8588-0198" }, { "id": "Wang-Jonathan-H", "name": { "family": "Wang", "given": "Jonathan H." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Looger-Loren-L", "name": { "family": "Looger", "given": "Loren L." }, "orcid": "0000-0002-7531-1757" }, { "id": "Gallagher-Timothy", "name": { "family": "Gallagher", "given": "Timothy" }, "orcid": "0000-0002-3544-327X" }, { "id": "Rees-D-C", "name": { "family": "Rees", "given": "Douglas C." }, "orcid": "0000-0003-4073-1185" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Fluorescence activation mechanism and imaging of drug permeation with new sensors for smoking-cessation ligands", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2022, Nichols et al. This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited. \n\nPreprinted: 04 October 2021; Received: 12 October 2021; Accepted: 03 January 2022; Published: 04 January 2022. \n\nWe thank Stefan Petrovic for his stewardship of the isothermal titration calorimeter in the Caltech Center for Molecular Medicine, Jens Kaiser for help with structural studies at the Caltech Molecular Observatory, the Gradinaru lab and Caltech CLOVER Center for help with viral vectors, and Andres Collazo and Giada Spigolon at the Caltech Biological Imaging Facility. We thank Zoe Beatty, Kallol Bera, Eve Fine, Shan Huang, Elaine Lin, Stephen Mayo, Lin Tian, and Elizabeth Unger for advice and guidance. We thank Achieve Life Sciences for a gift of cytisine. California Tobacco-Related Disease Research Program (TRDRP) (27 FT-0022), ALN. California Tobacco-Related Disease Research Program (TRDRP) (27IP-0057), HAL. California Tobacco-Related Disease Research Program (TRDRP) (T29IR0455), DAD. NIH (GM-123582, DA043829), HAL. NIH (DA049140, GM7616), AKM. Howard\nHughes Medical Institute (LLL, JSM, DCR). UK Engineering and Physical Sciences Research Council (no. EP/N024117/1), TG. Leiden University International Studies Fund (LISF L18020-1-45), LL. \n\nThe funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. \n\nAuthor contributions: Aaron L Nichols, Conceptualization, Data curation, Formal analysis, Funding acquisition, Investigation, Methodology, Supervision, Validation, Visualization, Writing \u2013 original draft, Writing \u2013 review and editing; Zack Blumenfeld, Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing \u2013 review and editing; Chengcheng Fan, Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing \u2013 original draft, Writing \u2013 review and editing; Laura Luebbert, Bruce N Cohen, Data curation, Formal analysis, Investigation, Methodology, Writing \u2013 review and editing; Annet EM Blom, Philip M Borden, Conceptualization, Investigation; Jonathan S Marvin, Data curation, Formal analysis, Investigation, Methodology, Validation, Writing \u2013 original draft, Writing \u2013 review and editing; Charlene H Kim, Data curation, Investigation, Methodology, Resources, Writing \u2013 review and editing; Anand K Muthusamy, Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Resources, Validation, Visualization; Amol V Shivange, Data curation, Validation; Hailey J Knox, Conceptualization, Methodology, Resources, Writing \u2013 review and editing; Hugo Rego Campello, Data curation, Resources, Validation; Jonathan H Wang, Investigation, Methodology, Resources; Dennis A Dougherty, Conceptualization, Data curation, Funding acquisition, Project administration, Resources, Supervision; Loren L Looger, Conceptualization, Funding acquisition, Project administration, Resources, Supervision, Writing \u2013 review and editing; Timothy Gallagher, Conceptualization, Funding acquisition, Project administration, Supervision, Writing \u2013 review and editing; Douglas C Rees, Data curation, Funding acquisition, Project administration, Supervision, Validation, Writing \u2013 review and editing; Henry A Lester, Conceptualization, Data curation, Formal analysis, Funding acquisition, Methodology, Project administration, Resources, Software, Supervision, Visualization, Writing \u2013 review and editing. \n\nData availability: The plasmids and associated database entries are available from Addgene (as named in our manuscript) with genetic maps. The Protein Data Bank has published the crystallographics and structural data (accession codes 7S7T, 7S7U, 7S7V). Supplementary File 1 gives relevant details.\n\nPublished - elife-74648-v2.pdf
Accepted Version - elife-74648-v1.pdf
Submitted - 2021.10.04.463082v4.full.pdf
Supplemental Material - elife-74648-supp1-v2.xlsx
Supplemental Material - elife-74648-supp2-v2.xlsx
Supplemental Material - elife-74648-transrepform1-v2.docx
", "abstract": "Nicotinic partial agonists provide an accepted aid for smoking cessation and thus contribute to decreasing tobacco-related disease. Improved drugs constitute a continued area of study. However, there remains no reductionist method to examine the cellular and subcellular pharmacokinetic properties of these compounds in living cells. Here, we developed new intensity-based drug-sensing fluorescent reporters (iDrugSnFRs) for the nicotinic partial agonists dianicline, cytisine, and two cytisine derivatives \u2013 10-fluorocytisine and 9-bromo-10-ethylcytisine. We report the first atomic-scale structures of liganded periplasmic binding protein-based biosensors, accelerating development of iDrugSnFRs and also explaining the activation mechanism. The nicotinic iDrugSnFRs detect their drug partners in solution, as well as at the plasma membrane (PM) and in the endoplasmic reticulum (ER) of cell lines and mouse hippocampal neurons. At the PM, the speed of solution changes limits the growth and decay rates of the fluorescence response in almost all cases. In contrast, we found that rates of membrane crossing differ among these nicotinic drugs by >30-fold. The new nicotinic iDrugSnFRs provide insight into the real-time pharmacokinetic properties of nicotinic agonists and provide a methodology whereby iDrugSnFRs can inform both pharmaceutical neuroscience and addiction neuroscience.", "date": "2022-01-04", "date_type": "published", "publication": "eLife", "volume": "11", "publisher": "eLife Sciences Publications", "pagerange": "Art. No. e74648", "id_number": "CaltechAUTHORS:20211008-224629724", "issn": "2050-084X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20211008-224629724", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "27FT-0022" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "27IP-0057" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "T29IR0455" }, { "agency": "NIH", "grant_number": "GM-123582" }, { "agency": "NIH", "grant_number": "DA043829" }, { "agency": "NIH", "grant_number": "DA049140" }, { "agency": "NIH", "grant_number": "GM7616" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "Engineering and Physical Sciences Research Council (EPSRC)", "grant_number": "EP/N024117/1" }, { "agency": "Leiden University", "grant_number": "LISF L18020-1-45" } ] }, "local_group": { "items": [ { "id": "Jacobs-Institute-for-Molecular-Engineering-for-Medicine" }, { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.7554/eLife.74648", "pmcid": "PMC8820738", "primary_object": { "basename": "elife-74648-transrepform1-v2.docx", "url": "https://authors.library.caltech.edu/records/ft222-e3n43/files/elife-74648-transrepform1-v2.docx" }, "related_objects": [ { "basename": "elife-74648-v1.pdf", "url": "https://authors.library.caltech.edu/records/ft222-e3n43/files/elife-74648-v1.pdf" }, { "basename": "elife-74648-v2.pdf", "url": "https://authors.library.caltech.edu/records/ft222-e3n43/files/elife-74648-v2.pdf" }, { "basename": "2021.10.04.463082v4.full.pdf", "url": "https://authors.library.caltech.edu/records/ft222-e3n43/files/2021.10.04.463082v4.full.pdf" }, { "basename": "elife-74648-supp1-v2.xlsx", "url": "https://authors.library.caltech.edu/records/ft222-e3n43/files/elife-74648-supp1-v2.xlsx" }, { "basename": "elife-74648-supp2-v2.xlsx", "url": "https://authors.library.caltech.edu/records/ft222-e3n43/files/elife-74648-supp2-v2.xlsx" } ], "resource_type": "article", "pub_year": "2022", "author_list": "Nichols, Aaron L.; Blumenfeld, Zack; et el." }, { "id": "https://authors.library.caltech.edu/records/azpps-zvb13", "eprint_id": 110334, "eprint_status": "archive", "datestamp": "2023-08-22 11:02:33", "lastmod": "2023-12-22 23:15:34", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Mulcahy-Matthew-J", "name": { "family": "Mulcahy", "given": "Matthew J." }, "orcid": "0000-0001-5588-8762" }, { "id": "Huard-Stephanie-M", "name": { "family": "Huard", "given": "Stephanie M." }, "orcid": "0000-0002-2333-1852" }, { "id": "Paulo-Joao-A", "name": { "family": "Paulo", "given": "Joao A." }, "orcid": "0000-0002-4291-413X" }, { "id": "Wang-Jonathan-H", "name": { "family": "Wang", "given": "Jonathan H." } }, { "id": "McKinney-Sheri-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Marks-Michael-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Henderson-Brandon-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Protein profiling in the habenula after chronic (\u2013)-menthol exposure in mice", "ispublished": "pub", "full_text_status": "public", "keywords": "addiction; immunoblotting; mass spectrometry; menthol; nicotine; radioligand binding", "note": "\u00a9 2021 International Society for Neurochemistry. \n\nIssue Online: 22 September 2021; Version of Record online: 02 September 2021; Accepted manuscript online: 18 August 2021; Manuscript accepted: 11 August 2021; Manuscript revised: 10 July 2021; Manuscript received: 02 March 2021. \n\nResearch Funding: National Institutes of Health. Grant Numbers: DA036061, DA040047, DK098285.\n\nAccepted Version - nihms-1734409.pdf
Supplemental Material - jnc15495-sup-0001-tables1.xlsx
", "abstract": "The identification of proteins that are altered following nicotine/tobacco exposure can facilitate and positively impact the investigation of related diseases. In this report, we investigated the effects of chronic (\u2013)-menthol exposure in 14 murine brain regions for changes in total \u03b22 subunit protein levels and changes in epibatidine binding levels using immunoblotting and radioligand binding assays. We identified the habenula as a region of interest due to the region's marked decreases in \u03b22 subunit and nAChR levels in response to chronic (\u2013)-menthol alone. Thus, we further examined the habenula, a brain region associated with both the reward and withdrawal components of addiction, for additional protein level alterations using mass spectrometry. A total of 552 proteins with altered levels were identified after chronic (\u2013)-menthol exposure. Enriched in the proteins with altered levels after (\u2013)-menthol exposure were proteins associated with signaling, immune systems, RNA regulation, and protein transport. The continuation and expansion of the brain region-specific protein profiling in response to (\u2013)-menthol will provide a better understanding of how this common flavorant in tobacco and e-liquid products may affect addiction and general health.", "date": "2021-09", "date_type": "published", "publication": "Journal of Neurochemistry", "volume": "158", "number": "6", "publisher": "Wiley", "pagerange": "1345-1358", "id_number": "CaltechAUTHORS:20210820-224447031", "issn": "0022-3042", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210820-224447031", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA036061" }, { "agency": "NIH", "grant_number": "DA040047" }, { "agency": "NIH", "grant_number": "DK098285" } ] }, "local_group": { "items": [ { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1111/jnc.15495", "pmcid": "PMC8577691", "primary_object": { "basename": "jnc15495-sup-0001-tables1.xlsx", "url": "https://authors.library.caltech.edu/records/azpps-zvb13/files/jnc15495-sup-0001-tables1.xlsx" }, "related_objects": [ { "basename": "nihms-1734409.pdf", "url": "https://authors.library.caltech.edu/records/azpps-zvb13/files/nihms-1734409.pdf" } ], "resource_type": "article", "pub_year": "2021", "author_list": "Mulcahy, Matthew J.; Huard, Stephanie M.; et el." }, { "id": "https://authors.library.caltech.edu/records/s7jfw-wxw93", "eprint_id": 109801, "eprint_status": "archive", "datestamp": "2023-08-20 04:17:08", "lastmod": "2023-12-22 23:31:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Grant-Stephen-N", "name": { "family": "Grant", "given": "Stephen N." }, "orcid": "0000-0003-0923-8886" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Regulation of epithelial sodium channel activity by SARS-CoV-1 and SARS-CoV-2 proteins", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2021 Biophysical Society. \n\nReceived 15 December 2020, Accepted 3 June 2021, Available online 29 June 2021. \n\nWe thank Drs. Christopher Barnes, Bruce N. Cohen, Douglas Eaton, Carolyn Machamer, Gerhard Thiel, Anthony West, Nael McCarty, Dennis A. Dougherty and Nathan Dascal for useful insights, and Jonathan Wang for harvesting oocytes. \n\nThis work was supported by the National Institute of Drug Abuse (DA046122 and DA049140), the National Institute of General Medical Sciences (GM-123582), and the California Tobacco-Related Disease Program (29IR0445 to D.A. Dougherty). \n\nAuthor contributions: S.N.G. designed and performed the experiments, analyzed the data, and prepared the manuscript. H.A.L. designed the experiments, analyzed the data, prepared the manuscript, and supervised the project.\n\nSupplemental Material - 1-s2.0-S0006349521004902-mmc1.pdf
", "abstract": "Severe acute respiratory syndrome (SARS) coronavirus (CoV) 2 (SARS-CoV-2), which causes the coronavirus disease 2019, encodes several proteins whose roles are poorly understood. We tested their ability either to directly form plasma membrane ion channels or to change functions of two mammalian plasma membrane ion channels, the epithelial sodium channel (ENaC) and the \u03b13\u03b24 nicotinic acetylcholine receptor. In mRNA-injected Xenopus oocytes, none of nine SARS-CoV-2 proteins or two SARS-CoV-1 proteins produced conductances, nor did co-injection of several combinations. Immunoblots for ORF8, spike (S), and envelope (E) proteins revealed that the proteins are expressed at appropriate molecular weights. In experiments on coexpression with ENaC, three tested SARS proteins (SARS-CoV-1 E, SARS-CoV-2 E, and SARS-CoV-2 S) markedly decrease ENaC currents. SARS-CoV-1 S protein decreases ENaC currents modestly. Coexpressing the E proteins but not the S proteins with \u03b13\u03b24 nicotinic acetylcholine receptors significantly reduces acetylcholine-induced currents. ENaC inhibition does not occur if the SARS-CoV protein mRNAs are injected 24 h after the ENaC mRNAs, suggesting that SARS-CoV proteins affect early step(s) in functional expression of channel proteins. Consistent with the hypothesis that the SARS-CoV-2 S protein-induced ENaC inhibition involves competition for available protease, mutating the furin cleavage site in SARS-CoV-2 S protein partially relieves inhibition of ENaC currents. Extending previous suggestions that SARS proteins affect ENaC currents via protein kinase C (PKC) activation, PKC activation via phorbol 12-myristate 13-acetate decreases ENaC and \u03b13\u03b24 activity. Phorbol 12-myristate 13-acetate application reduced membrane capacitance \u223c5%, presumably via increased endocytosis, but this decrease is much smaller than the SARS proteins' effects on conductances. Also, incubating oocytes in G\u00f6-6976, a PKC\u03b1 and PKC\u03b2 inhibitor, did not alter E or S protein-induced channel inhibition. We conclude that SARS-CoV-1 and SARS-CoV-2 proteins alter the function of human plasma membrane channels, via incompletely understood mechanisms. These interactions may play a role in the coronavirus 2019 pathophysiology.", "date": "2021-07-20", "date_type": "published", "publication": "Biophysical Journal", "volume": "120", "number": "14", "publisher": "Biophysical Society", "pagerange": "2805-2813", "id_number": "CaltechAUTHORS:20210714-140716932", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210714-140716932", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Institute on Drug Abuse" }, { "agency": "NIH", "grant_number": "DA046122" }, { "agency": "NIH", "grant_number": "DA049140" }, { "agency": "NIH", "grant_number": "GM-123582" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "29IR0445" } ] }, "local_group": { "items": [ { "id": "COVID-19" }, { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1016/j.bpj.2021.06.005", "pmcid": "PMC8238646", "primary_object": { "basename": "1-s2.0-S0006349521004902-mmc1.pdf", "url": "https://authors.library.caltech.edu/records/s7jfw-wxw93/files/1-s2.0-S0006349521004902-mmc1.pdf" }, "resource_type": "article", "pub_year": "2021", "author_list": "Grant, Stephen N. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/hjhgm-hc812", "eprint_id": 107173, "eprint_status": "archive", "datestamp": "2023-08-22 08:05:21", "lastmod": "2023-12-22 23:35:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Unger-Elizabeth-K", "name": { "family": "Unger", "given": "Elizabeth K." }, "orcid": "0000-0002-8235-7534" }, { "name": { "family": "Keller", "given": "Jacob P." }, "orcid": "0000-0001-7487-4104" }, { "id": "Altermatt-Michael", "name": { "family": "Altermatt", "given": "Michael" }, "orcid": "0000-0003-2841-5374" }, { "name": { "family": "Liang", "given": "Ruqiang" }, "orcid": "0000-0002-3075-4554" }, { "name": { "family": "Matsui", "given": "Aya" }, "orcid": "0000-0003-4437-8278" }, { "name": { "family": "Dong", "given": "Chunyang" }, "orcid": "0000-0002-4820-4454" }, { "name": { "family": "Hon", "given": "Olivia J." }, "orcid": "0000-0003-1086-1421" }, { "name": { "family": "Yao", "given": "Zizhen" }, "orcid": "0000-0002-9361-5607" }, { "name": { "family": "Sun", "given": "Junqing" } }, { "name": { "family": "Banala", "given": "Sambashiva" }, "orcid": "0000-0002-9463-7664" }, { "name": { "family": "Flanigan", "given": "Meghan E." }, "orcid": "0000-0002-3185-7459" }, { "name": { "family": "Jaffe", "given": "David A." }, "orcid": "0000-0003-4773-6982" }, { "name": { "family": "Hartanto", "given": "Samantha" }, "orcid": "0000-0001-8513-5294" }, { "name": { "family": "Carlen", "given": "Jane" }, "orcid": "0000-0002-2538-6670" }, { "name": { "family": "Mizuno", "given": "Grace O." }, "orcid": "0000-0003-4786-3084" }, { "name": { "family": "Borden", "given": "Phillip M." }, "orcid": "0000-0003-1653-7067" }, { "id": "Shivange-Amol-V", "name": { "family": "Shivange", "given": "Amol V." }, "orcid": "0000-0002-4169-2969" }, { "name": { "family": "Cameron", "given": "Lindsay P." }, "orcid": "0000-0002-8420-7898" }, { "name": { "family": "Sinning", "given": "Steffen" }, "orcid": "0000-0001-6971-6929" }, { "name": { "family": "Underhill", "given": "Suzanne M." } }, { "name": { "family": "Olson", "given": "David E." }, "orcid": "0000-0002-4517-0543" }, { "name": { "family": "Amara", "given": "Susan G." }, "orcid": "0000-0001-8914-1106" }, { "name": { "family": "Temple Lang", "given": "Duncan" }, "orcid": "0000-0003-0159-1546" }, { "name": { "family": "Rudnick", "given": "Gary" }, "orcid": "0000-0002-7622-4110" }, { "name": { "family": "Marvin", "given": "Jonathan S." }, "orcid": "0000-0003-2294-4515" }, { "name": { "family": "Lavis", "given": "Luke D." }, "orcid": "0000-0002-0789-6343" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "name": { "family": "Alvarez", "given": "Veronica A." }, "orcid": "0000-0003-2611-8675" }, { "name": { "family": "Fisher", "given": "Andrew J." }, "orcid": "0000-0003-3488-6594" }, { "name": { "family": "Prescher", "given": "Jennifer A." }, "orcid": "0000-0002-9250-4702" }, { "name": { "family": "Kash", "given": "Thomas L." }, "orcid": "0000-0002-4747-4495" }, { "name": { "family": "Yarov-Yarovoy", "given": "Vladimir" }, "orcid": "0000-0002-2325-4834" }, { "id": "Gradinaru-V", "name": { "family": "Gradinaru", "given": "Viviana" }, "orcid": "0000-0001-5868-348X" }, { "name": { "family": "Looger", "given": "Loren L." }, "orcid": "0000-0002-7531-1757" }, { "name": { "family": "Tian", "given": "Lin" }, "orcid": "0000-0001-7012-6926" } ] }, "title": "Directed Evolution of a Selective and Sensitive Serotonin Sensor via Machine Learning", "ispublished": "pub", "full_text_status": "public", "keywords": "serotonin; fluorescence protein sensor; fiber photometry; machine learning; iSeroSnFR; SERT; OSTA; sleep-wake; fear-learning; social behaviors", "note": "\u00a9 2020 Elsevier. \n\nReceived 23 November 2019, Revised 22 June 2020, Accepted 20 November 2020, Available online 16 December 2020. \n\nWe would like to thank Drs. Liqun Luo (Stanford University) and Jing Ren (MRC) for their critical reading and feedback. This work is based upon research conducted at the Northeastern Collaborative Access Team beamlines, which are funded by the National Institute of General Medical Sciences from the NIH (P30-GM124165). The Pilatus 6M detector on the 24-ID-C beamline is funded by an NIH-ORIP HEI grant (S10-RR029205). This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract DE-AC02-06CH11357. This work was supported by funding to L.T. (BRAIN Initiative U01NS090604, U01NS013522, and DP2MH107056 from NIH), to E.K.U. (Mistletoe Foundation Research Fellowship), and to G.O.M. (ARCS Scholarship), as well as by the Howard Hughes Medical Institute. V.G. is a Heritage Principal Investigator supported by the Heritage Medical Research Institute, the NIH (BRAIN RF1MH117069), the Center for Molecular and Cellular Neuroscience of the Chen Institute, and the Beckman Institute for CLARITY, Optogenetics and Vector Engineering Research. V.A.A. is funded by Intramural Programs of NIAAA and NINDS ZIA-AA000421 and Innovation Award from NIH-DDIR. T.L.K. is funded by the NIH (R01AA019454, P60AA011605, and U24AA025475), and O.J.H. is funded by the NIH (5T32NS007431-20). \n\nAuthor Contributions. E.K.U., J.P.K., M.A., L.L.L., and L.T. conceived of and designed the study. E.K.U. designed the machine-learning method, screened and optimized sensors, and characterized them in purified protein, mammalian cells, cultured neurons, and brain slice, with significant contribution from C.D., D.A.J., and J.S. J.P.K., S.S., and G.R. designed OSTA and stopped-flow experiments, and J.P.K. performed them. M.A. and V.G. designed and performed in vivo fiber photometry and EEG/EMG recording in BLA and mPFC in fear learning and sleep/wake cycles. O.J.H., M.E.F., and T.L.K. designed and performed in vivo fiber photometry experiments in BLA, OFC, and BNST during social interaction. R.L. and V.Y.-Y. designed and performed computational Rosetta modeling. Z.Y. and J.A.P. provided luciferase experimental data for establishing machine-learning methods. J.C. and D.T.L. provided significant insight for the machine-learning methods. J.S. characterized the sensor in acute slice using two-photon imaging. A.M. and V.A.A. designed and performed photometry imaging in acute slice. S.H. and A.J.F. performed crystallography. J.S.M., P.M.B., A.V.S., H.A.L., and L.L.L. provided iAChSnFR0.6 and performed preliminary experiments on serotonin binding. S.B. and L.D.L. synthesized caged serotonin. G.O.M. provided dissociated neuronal cultures. L.P.C. and D.E.O. produced chemical reagents. S.M.U. and S.G.A. provided SSRIs and guidance in cell-assay design. E.K.U., J.P.K., L.L.L., and L.T. wrote the manuscript with significant input from other authors. \n\nDeclaration of Interests. L.T. and G.O.M. are co-founders of Seven Biosciences. D.E.O. is a founder of Delix.\n\nAccepted Version - nihms-1654804.pdf
Supplemental Material - 1-s2.0-S0092867420316123-mmc1.xlsx
Supplemental Material - 1-s2.0-S0092867420316123-mmc2.pdf
", "abstract": "Serotonin plays a central role in cognition and is the target of most pharmaceuticals for psychiatric disorders. Existing drugs have limited efficacy; creation of improved versions will require better understanding of serotonergic circuitry, which has been hampered by our inability to monitor serotonin release and transport with high spatial and temporal resolution. We developed and applied a binding-pocket redesign strategy, guided by machine learning, to create a high-performance, soluble, fluorescent serotonin sensor (iSeroSnFR), enabling optical detection of millisecond-scale serotonin transients. We demonstrate that iSeroSnFR can be used to detect serotonin release in freely behaving mice during fear conditioning, social interaction, and sleep/wake transitions. We also developed a robust assay of serotonin transporter function and modulation by drugs. We expect that both machine-learning-guided binding-pocket redesign and iSeroSnFR will have broad utility for the development of other sensors and in vitro and in vivo serotonin detection, respectively.", "date": "2020-12-23", "date_type": "published", "publication": "Cell", "volume": "183", "number": "7", "publisher": "Cell Press", "pagerange": "1986-2002", "id_number": "CaltechAUTHORS:20201217-143607354", "issn": "0092-8674", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201217-143607354", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "P30-GM124165" }, { "agency": "NIH", "grant_number": "S10-RR029205" }, { "agency": "Department of Energy (DOE)", "grant_number": "DE-AC02-06CH11357" }, { "agency": "NIH", "grant_number": "U01NS090604" }, { "agency": "NIH", "grant_number": "U01NS013522" }, { "agency": "NIH", "grant_number": "DP2MH107056" }, { "agency": "Mistletoe Foundation" }, { "agency": "ARCS Foundation" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "Heritage Medical Research Institute" }, { "agency": "NIH", "grant_number": "RF1MH117069" }, { "agency": "Tianqiao and Chrissy Chen Institute for Neuroscience" }, { "agency": "Caltech Beckman Institute" }, { "agency": "NIH", "grant_number": "ZIA-AA000421" }, { "agency": "NIH", "grant_number": "R01AA019454" }, { "agency": "NIH", "grant_number": "P60AA011605" }, { "agency": "NIH", "grant_number": "U24AA025475" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5T32NS007431-20" } ] }, "local_group": { "items": [ { "id": "Heritage-Medical-Research-Institute" }, { "id": "Tianqiao-and-Chrissy-Chen-Institute-for-Neuroscience" }, { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1016/j.cell.2020.11.040", "pmcid": "PMC8025677", "primary_object": { "basename": "1-s2.0-S0092867420316123-mmc1.xlsx", "url": "https://authors.library.caltech.edu/records/hjhgm-hc812/files/1-s2.0-S0092867420316123-mmc1.xlsx" }, "related_objects": [ { "basename": "1-s2.0-S0092867420316123-mmc2.pdf", "url": "https://authors.library.caltech.edu/records/hjhgm-hc812/files/1-s2.0-S0092867420316123-mmc2.pdf" }, { "basename": "nihms-1654804.pdf", "url": "https://authors.library.caltech.edu/records/hjhgm-hc812/files/nihms-1654804.pdf" } ], "resource_type": "article", "pub_year": "2020", "author_list": "Unger, Elizabeth K.; Keller, Jacob P.; et el." }, { "id": "https://authors.library.caltech.edu/records/zsn9f-73t54", "eprint_id": 106314, "eprint_status": "archive", "datestamp": "2023-08-20 00:05:33", "lastmod": "2023-12-22 23:15:30", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Grant-Stephen", "name": { "family": "Grant", "given": "Stephen" }, "orcid": "0000-0003-0923-8886" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Proteins for increased surface expression of the \u03b16\u03b24 nicotinic acetylcholine receptor: Nothing but good news?", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2020 American Society for Clinical Investigation. \n\nPublished October 19, 2020. \n\nSG and HAL are supported in part by NIH grants DA046122, GM123582, and MH120823. \n\nThe authors have declared that no conflict of interest exists.\n\nPublished - jci-130-143197.pdf
", "abstract": "Useful animal models of disease in neuroscience can make accurate predictions about a therapeutic outcome, a feature known as predictive validity. In this issue of the JCI, Knowland et al. provide an improved model to assess nicotinic acetylcholine receptor (nAChR) ligands for treating chronic pain. The authors identify two proteins, the voltage-dependent calcium channel auxiliary subunit BARP and the unfolded protein response sensor IRE1\u03b1, that are required for robust heterologous expression of \u03b16\u03b24, an nAChR subtype in dorsal root ganglia (DRG). This nAChR is a candidate for the analgesic effects of nicotine as well as the frog toxin epibatidine. Now researchers can efficiently screen for \u03b16\u03b24 nAChR\u2013selective agonists using heterologous expression systems. Candidates that emerge will enable researchers to test the predictive validity of mouse models for chronic pain in the nAChR context. If all these steps work, one can envision a class of non-opioid nAChR-targeted analgesics for chronic pain.", "date": "2020-11", "date_type": "published", "publication": "Journal of Clinical Investigation", "volume": "130", "number": "11", "publisher": "American Society for Clinical Investigation", "pagerange": "5685-5687", "id_number": "CaltechAUTHORS:20201028-074338126", "issn": "0021-9738", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201028-074338126", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA046122" }, { "agency": "NIH", "grant_number": "GM123582" }, { "agency": "NIH", "grant_number": "MH120823" } ] }, "local_group": { "items": [ { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1172/jci143197", "pmcid": "PMC7598034", "primary_object": { "basename": "jci-130-143197.pdf", "url": "https://authors.library.caltech.edu/records/zsn9f-73t54/files/jci-130-143197.pdf" }, "resource_type": "article", "pub_year": "2020", "author_list": "Grant, Stephen and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/g7yqa-9qd26", "eprint_id": 101193, "eprint_status": "archive", "datestamp": "2023-08-19 19:54:20", "lastmod": "2023-12-22 23:15:36", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Grant-S", "name": { "family": "Grant", "given": "Stephen" }, "orcid": "0000-0003-0923-8886" }, { "id": "Muthusamy-A-K", "name": { "family": "Muthusamy", "given": "Anand K." }, "orcid": "0000-0003-1041-914X" }, { "id": "Collazo-A", "name": { "family": "Collazo", "given": "Andres" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Antagonists Pharmacologically Chaperone Opioid Receptors", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2020 Biophysical Society. \n\nAvailable online 7 February 2020. \n\nSupport: DA049140, DA046122.", "abstract": "More than 100 people die daily in the United States from opioid-related drug overdoses. \u00b5-Opioid receptor (MOR) antagonists, such as naltrexone (Ntx) and naloxone, partially suppress overdose effects, but also induce supersensitivity to MOR activation. Opioid receptors are folded in the endoplasmic reticulum (ER), undergo transport to the Golgi, eventually reach the plasma membrane, and may also later undergo endocytosis. We examined the effects of opioid ligands on MOR trafficking during the earlier processes. Properly folded protein cargo recruits ER exit sites (ERES) and then enters coated vesicles for delivery to the Golgi. We transfected SH-SY5Y cells with fluorescently tagged Sec24 variants to visualize and quantify ERES, and with the strongly ER-retained MOR mutant, MOR[N190K]. We attained sub-\u00b5m resolution, for information on ligand effects on ERES levels in live cells. F\u00f6rster resonance energy transfer (FRET) showed that MOR closely interacts with both Sec24C and Sec24D. Using Sec24D-eGFP to fluorescently mark ERES, we observed that the antagonists increase the fraction of the cytoplasm occupied by ERES. We also found that SH-SY5Y cells overexpressing wild-type \u03b4-opioid receptors have increased ERES levels after exposure to naloxone. These effects most likely occur when an antagonist acts as a pharmacological chaperone of opioid receptors. We rule out the alternative hypothesis that antagonists affect ERES levels via changes in [cAMP] because Ntx did not detectably change [cAMP]. In contrast to these effects of antagonists, several opioid agonists (morphine, fentanyl, buprenorphine, and methadone) lacked detectable effects on ERES levels. Full or partial agonists, but not antagonists, phosphorylate MOR at S375, but SH-SY5Y cells overexpressing MOR[N190K][S375A] showed no change in ERES density in response to agonists. The possibility that antagonists induce supersensitivity by pharmacologically chaperoning opioid receptors could suggest innovative approaches for opioid abuse disorder.", "date": "2020-02-07", "date_type": "published", "publication": "Biophysical Journal", "volume": "118", "number": "3", "publisher": "Biophysical Society", "pagerange": "27a", "id_number": "CaltechAUTHORS:20200210-091504115", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200210-091504115", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA049140" }, { "agency": "NIH", "grant_number": "DA046122" } ] }, "local_group": { "items": [ { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1016/j.bpj.2019.11.328", "resource_type": "article", "pub_year": "2020", "author_list": "Grant, Stephen; Muthusamy, Anand K.; et el." }, { "id": "https://authors.library.caltech.edu/records/z9q5a-d3f28", "eprint_id": 99486, "eprint_status": "archive", "datestamp": "2023-08-19 19:28:50", "lastmod": "2023-12-22 23:15:28", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Mulcahy-M-J", "name": { "family": "Mulcahy", "given": "Matthew J." }, "orcid": "0000-0001-5588-8762" }, { "id": "Huard-S-M", "name": { "family": "Huard", "given": "Stephanie M." }, "orcid": "0000-0002-2333-1852" }, { "id": "Paulo-J-A", "name": { "family": "Paulo", "given": "Joao A." }, "orcid": "0000-0002-4291-413X" }, { "id": "Wang-Jonathan-H", "name": { "family": "Wang", "given": "Jonathan H." } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Brain Region-Specific nAChR and Associated Protein Abundance Alterations Following Chronic Nicotine and/or Menthol Exposure", "ispublished": "pub", "full_text_status": "public", "keywords": "\u03b22 nAChR subunit, nicotine, menthol, immunoblotting, mass spectrometry, addiction", "note": "\u00a9 2019 American Chemical Society. \n\nReceived: April 30, 2019; Published: October 28, 2019. \n\nWe also thank Dr. Steven P. Gygi and the Taplin Mass Spectrometry Facility at Harvard Medical School for use of their mass spectrometers and access to their Sequest-based data analysis suite. \n\nAuthor Contributions: The manuscript was written through the contributions of all authors. All authors have given approval to the final version of the manuscript. M.J.M., S.M.H., J.A.P., B.J.H., and H.A.L. conceived and designed experiments. M.J.M., S.M.H., and J.A.P. performed the experiments. J.A.P., J.H.W., and S.M. contributed reagents/materials/analysis tools. M.J.M. wrote the manuscript. M.J.M., S.M.H., J.A.P., B.J.H., and H.A.L. edited the manuscript. \n\nThis research was supported by NIH R01DA036061 (H.A.L.) and R01GM132129 (J.A.P.). \n\nThe authors declare no competing financial interest. \n\nThe mass spectrometry proteomics data have been deposited to the PRIDE Archive (http://www.ebi.ac.uk/pride/archive/) via the PRIDE partner repository with the data set identifier PXD012244.(69)\n\nAccepted Version - nihms-1595991.pdf
Supplemental Material - pr9b00286_si_001.xlsx
", "abstract": "The identification of biomarkers that are altered following nicotine/tobacco exposure can facilitate the investigation of tobacco-related diseases. Nicotinic acetylcholine receptors (nAChRs) are pentameric cation channels expressed in the mammalian central and peripheral nervous systems and the neuromuscular junction. Neuronal nAChR subunits (11) have been identified in mammals (\u03b12-7, \u03b19-10, \u03b22-4). We examined changes in \u03b22 nAChR subunit protein levels after chronic nicotine, (\u00b1)-menthol, or nicotine co-administered with (\u00b1)-menthol in nine murine brain regions. Our investigation of \u03b22 nAChR subunit level changes identified the hypothalamus as a novel region of interest for menthol exposure that demonstrated increased \u03b22 nAChR levels after (\u00b1)-menthol plus nicotine exposure compared to nicotine exposure alone. Using mass spectrometry, we further characterized changes in membrane protein abundance profiles in the hypothalamus to identify potential biomarkers of (\u00b1)-menthol plus nicotine exposure and proteins that may contribute to the elevated \u03b22 nAChR subunit levels. In the hypothalamus, 272 membrane proteins were identified with altered abundances after chronic nicotine plus menthol exposure with respect to chronic nicotine exposure without menthol. A comprehensive investigation of changes in nAChR and non-nAChR protein expression resulting from (\u00b1)-menthol plus nicotine in the brain may establish biomarkers to better understand the effects of these drugs on addiction and addiction-related diseases.", "date": "2020-01-03", "date_type": "published", "publication": "Journal of Proteome Research", "volume": "19", "number": "1", "publisher": "American Chemical Society", "pagerange": "36-48", "id_number": "CaltechAUTHORS:20191028-111942058", "issn": "1535-3893", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191028-111942058", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "R01DA036061" }, { "agency": "NIH", "grant_number": "R01GM132129" } ] }, "local_group": { "items": [ { "id": "Division-of-Biology-and-Biological-Engineering" } ] }, "doi": "10.1021/acs.jproteome.9b00286", "pmcid": "PMC7289315", "primary_object": { "basename": "nihms-1595991.pdf", "url": "https://authors.library.caltech.edu/records/z9q5a-d3f28/files/nihms-1595991.pdf" }, "related_objects": [ { "basename": "pr9b00286_si_001.xlsx", "url": "https://authors.library.caltech.edu/records/z9q5a-d3f28/files/pr9b00286_si_001.xlsx" } ], "resource_type": "article", "pub_year": "2020", "author_list": "Mulcahy, Matthew J.; Huard, Stephanie M.; et el." }, { "id": "https://authors.library.caltech.edu/records/50jch-tz287", "eprint_id": 100341, "eprint_status": "archive", "datestamp": "2023-08-19 18:42:59", "lastmod": "2023-10-18 19:52:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Bera-K", "name": { "family": "Bera", "given": "Kallol" } }, { "id": "Kamajaya-A", "name": { "family": "Kamajaya", "given": "Aron" } }, { "id": "Shivange-A-V", "name": { "family": "Shivange", "given": "Amol V." } }, { "id": "Muthusamy-A-K", "name": { "family": "Muthusamy", "given": "Anand K." }, "orcid": "0000-0003-1041-914X" }, { "id": "Nichols-A-L", "name": { "family": "Nichols", "given": "Aaron L." }, "orcid": "0000-0001-9341-0049" }, { "id": "Borden-P-M", "name": { "family": "Borden", "given": "Philip M." } }, { "id": "Grant-S", "name": { "family": "Grant", "given": "Stephen" }, "orcid": "0000-0003-0923-8886" }, { "id": "Jeon-Janice-H", "name": { "family": "Jeon", "given": "Janice" }, "orcid": "0000-0002-8483-586X" }, { "id": "Lin-Elaine", "name": { "family": "Lin", "given": "Elaine" } }, { "id": "Bishara-I", "name": { "family": "Bishara", "given": "Ishak" }, "orcid": "0000-0001-7563-1980" }, { "id": "Chin-Theodore-M", "name": { "family": "Chin", "given": "Theodore M." } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "Unger-E-K", "name": { "family": "Unger", "given": "Elizabeth K." } }, { "id": "Tian-Lin", "name": { "family": "Tian", "given": "Lin" }, "orcid": "0000-0001-7012-6926" }, { "id": "Marvin-J-S", "name": { "family": "Marvin", "given": "Jonathan S." }, "orcid": "0000-0003-2294-4515" }, { "id": "Looger-L-L", "name": { "family": "Looger", "given": "Loren L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Biosensors Show the Pharmacokinetics of S-Ketamine in the Endoplasmic Reticulum", "ispublished": "pub", "full_text_status": "public", "keywords": "antidepressants, organelles, green fluorescent protein, protein engineering and design, periplasmic binding proteins (PBPs), inside-out pharmacology, iSketSnFR1, iSketSnFR2", "note": "\u00a9 2019 Bera, Kamajaya, Shivange, Muthusamy, Nichols, Borden, Grant, Jeon, Lin, Bishara, Chin, Cohen, Kim, Unger, Tian, Marvin, Looger and Lester. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. \n\nReceived: 19 August 2019; Accepted: 22 October 2019;\nPublished: 12 November 2019. \n\nData Availability Statement: The datasets generated for this study are available on request to the corresponding author. \n\nAuthor Contributions: KB, AK, AS, PB, IB, TC, AM, SG, CK and JM: performed experiments. KB, AK, AS, AM, AN, JJ, EL, BC, JM and HL: analysis. BC, JM, LL and HL: research direction. EU and LT: constructs. AN, BC, KB, EL, LL and HL: manuscript preparation and revision. LL, KB and HL: funding. \n\nThis research was supported by grants from US National Institutes of Health (GM123582, MH120823, DA046122, NS090604, NS013522, MH107056), the California Institute for Regenerative Medicine (EDUC2-08398), the Brain and Behavior Research Foundation (NARSAD), the Della Martin Foundation, the Howard Hughes Medical Institute, the Caltech CI2 program, Caltech SURF donors David and Karen Rossum, and the Mistletoe Foundation.\n\nConflict of Interest: LT is the founder of Seven Biosciences. \n\nThe remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. \n\nJacob P. Keller: advice on pH and tubing. Laura Luebbert: help with experiments. Luke L. Lavis: synthesis of N,N-dimethyl-S-ketamine. Michael Maher: advice on ketamine. Anindya Bhattacharya: advice on ketamine. Daniel Wagenaar: construction of LED light sources. Lauren M. Barnett: advice on photochemistry. Eric R. Schreiter: biosensors. Jonathan Wang: technical help. Margaret Jefferies and Purnima Deshpande: excellent lab management at Janelia and Caltech.\n\nPublished - fncel-13-00499.pdf
Supplemental Material - Presentation_1_Biosensors_Show_the_Pharmacokinetics_of_S-Ketamine_in_the_Endoplasmic_Reticulum.pdf
", "abstract": "The target for the \"rapid\" (<24 h) antidepressant effects of S-ketamine is unknown, vitiating programs to rationally develop more effective rapid antidepressants. To describe a drug's target, one must first understand the compartments entered by the drug, at all levels\u2014the organ, the cell, and the organelle. We have, therefore, developed molecular tools to measure the subcellular, organellar pharmacokinetics of S-ketamine. The tools are genetically encoded intensity-based S-ketamine-sensing fluorescent reporters, iSKetSnFR1 and iSKetSnFR2. In solution, these biosensors respond to S-ketamine with a sensitivity, S-slope = delta(F/F0)/(delta[S-ketamine]) of 0.23 and 1.9/\u03bcM, respectively. The iSKetSnFR2 construct allows measurements at <0.3 \u03bcM S-ketamine. The iSKetSnFR1 and iSKetSnFR2 biosensors display >100-fold selectivity over other ligands tested, including R-ketamine. We targeted each of the sensors to either the plasma membrane (PM) or the endoplasmic reticulum (ER). Measurements on these biosensors expressed in Neuro2a cells and in human dopaminergic neurons differentiated from induced pluripotent stem cells (iPSCs) show that S-ketamine enters the ER within a few seconds after appearing in the external solution near the PM, then leaves as rapidly after S-ketamine is removed from the extracellular solution. In cells, S-slopes for the ER and PM-targeted sensors differ by <2-fold, indicating that the ER [S-ketamine] is less than 2-fold different from the extracellular [S-ketamine]. Organelles represent potential compartments for the engagement of S-ketamine with its antidepressant target, and potential S-ketamine targets include organellar ion channels, receptors, and transporters.", "date": "2019-11-12", "date_type": "published", "publication": "Frontiers in Cellular Neuroscience", "volume": "13", "publisher": "Frontiers Research Foundation", "pagerange": "Art. No. 499", "id_number": "CaltechAUTHORS:20191218-090117543", "issn": "1662-5102", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191218-090117543", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM123582" }, { "agency": "NIH", "grant_number": "MH120823" }, { "agency": "NIH", "grant_number": "DA046122" }, { "agency": "NIH", "grant_number": "NS090604" }, { "agency": "NIH", "grant_number": "NS013522" }, { "agency": "NIH", "grant_number": "MH107056" }, { "agency": "California Institute for Regenerative Medicine (CIRM)", "grant_number": "EDUC2-08398" }, { "agency": "Brain and Behavior Research Foundation" }, { "agency": "Della Martin Foundation" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "Caltech Innovation Initiative (CI2)" }, { "agency": "Caltech Summer Undergraduate Research Fellowship (SURF)" }, { "agency": "Mistletoe Foundation" } ] }, "doi": "10.3389/fncel.2019.00499", "pmcid": "PMC6874132", "primary_object": { "basename": "Presentation_1_Biosensors_Show_the_Pharmacokinetics_of_S-Ketamine_in_the_Endoplasmic_Reticulum.pdf", "url": "https://authors.library.caltech.edu/records/50jch-tz287/files/Presentation_1_Biosensors_Show_the_Pharmacokinetics_of_S-Ketamine_in_the_Endoplasmic_Reticulum.pdf" }, "related_objects": [ { "basename": "fncel-13-00499.pdf", "url": "https://authors.library.caltech.edu/records/50jch-tz287/files/fncel-13-00499.pdf" } ], "resource_type": "article", "pub_year": "2019", "author_list": "Bera, Kallol; Kamajaya, Aron; et el." }, { "id": "https://authors.library.caltech.edu/records/j4tdc-kqs64", "eprint_id": 99686, "eprint_status": "archive", "datestamp": "2023-08-19 18:40:41", "lastmod": "2023-10-18 18:40:09", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Nicotine Bound to Its Receptors: New Structures for a Vexing Pathopharmacological Problem", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2019 Published by Elsevier Inc. \n\nAvailable online 6 November 2019.", "abstract": "The paper in this issue of Neuron (Gharpure et al., 2019), on the structure of nicotine bound to the \u03b13\u03b24 nicotinic acetylcholine receptor, provides important information that helps to explain the patterns of nicotine ingestion. Let's elaborate. Despite the heavy metabolic cost, various plant species produce molecules that are toxic to nervous systems. In many cases, these toxins discourage invertebrate herbivores.", "date": "2019-11-06", "date_type": "published", "publication": "Neuron", "volume": "104", "number": "3", "publisher": "Cell Press", "pagerange": "431-432", "id_number": "CaltechAUTHORS:20191106-094121720", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191106-094121720", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/j.neuron.2019.10.029", "resource_type": "article", "pub_year": "2019", "author_list": "Lester, Henry A. and Dougherty, Dennis A." }, { "id": "https://authors.library.caltech.edu/records/2ekzs-dxp63", "eprint_id": 98658, "eprint_status": "archive", "datestamp": "2023-08-19 18:15:15", "lastmod": "2023-10-18 17:32:26", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Blom-A-E-M", "name": { "family": "Blom", "given": "Annet E. M." }, "orcid": "0000-0002-7441-4893" }, { "id": "Rego-Campello-H", "name": { "family": "Rego Campello", "given": "Hugo" }, "orcid": "0000-0001-8588-0198" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Gallagher-T", "name": { "family": "Gallagher", "given": "Timothy" }, "orcid": "0000-0002-3544-327X" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Probing Binding Interactions of Cytisine Derivatives to the \u03b14\u03b22 Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2019 American Chemical Society. \n\nReceived: June 20, 2019; Published: September 13, 2019. \n\nWe thank Dr. Aurelian Honraedt for help in the synthesis of the derivatives. We thank Dr. Justin A. Hilf and Prof. Brian Stoltz for their assistance in calculating logD values. We thank Achieve Life Sciences for their generous gift of (\u2212)-cytisine, and the University of Bristol and Engineering and Physical Sciences Research Council (No. EP/N024117/1) for financial support. This work was partially supported by funds provided by The Regents of the University of California, Research Grants Program Office, Tobacco Related Disease Research Program (Grant No. T29IR0445). The opinions, findings, and conclusions herein are those of the authors and do not necessarily represent those of The Regents of the University of California, or any of its programs. \n\nAuthor Contributions: All authors have revised and given approval to the final version of the manuscript. \n\nThe authors declare no competing financial interest.\n\nSupplemental Material - ja9b06580_si_001.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are crucial for communication between synapses in the central nervous system. As such, they are also implicated in several neuropsychiatric and addictive diseases. Cytisine is a partial agonist of some nAChRs and has been used for smoking cessation. Previous studies have established a binding model for several agonists to several nAChR subtypes. Here, we evaluate the extent to which this model applies to cytisine at the \u03b14\u03b22 nAChR, which is a subtype that is known to play a prominent role in nicotine addiction. Along with the commonly seen cation\u2212\u03c0 interaction and two hydrogen bonds, we find that cytisine makes a second cation\u2212\u03c0 interaction at the agonist binding site. We also evaluated a series of C(10)-substituted cytisine derivatives, using two-electrode voltage-clamp electrophysiology and noncanonical amino acid mutagenesis. Double-mutant cycle analyses revealed that C(10) substitution generally strengthens the newly established second cation\u2212\u03c0 interaction, while it weakens the hydrogen bond typically seen to LeuE in the complementary subunit. The results suggest a model for how cytisine derivatives substituted at C(10) (as well as C(9)/C(10)) adjust their binding orientation, in response to pyridone ring substitution.", "date": "2019-10-09", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "141", "number": "40", "publisher": "American Chemical Society", "pagerange": "15840-15849", "id_number": "CaltechAUTHORS:20190916-135720189", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190916-135720189", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "University of Bristol" }, { "agency": "Engineering and Physical Sciences Research Council (EPSRC)", "grant_number": "EP/N024117/1" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "T29IR0445" } ] }, "doi": "10.1021/jacs.9b06580", "primary_object": { "basename": "ja9b06580_si_001.pdf", "url": "https://authors.library.caltech.edu/records/2ekzs-dxp63/files/ja9b06580_si_001.pdf" }, "resource_type": "article", "pub_year": "2019", "author_list": "Blom, Annet E. M.; Rego Campello, Hugo; et el." }, { "id": "https://authors.library.caltech.edu/records/xmy9t-8yy14", "eprint_id": 92625, "eprint_status": "archive", "datestamp": "2023-08-19 16:00:02", "lastmod": "2023-10-20 15:55:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Shivange-Amol-V", "name": { "family": "Shivange", "given": "Amol V." }, "orcid": "0000-0002-4169-2969" }, { "id": "Borden-Philip-M", "name": { "family": "Borden", "given": "Philip M." } }, { "id": "Muthusamy-Anand-K", "name": { "family": "Muthusamy", "given": "Anand K." }, "orcid": "0000-0003-1041-914X" }, { "id": "Nichols-Aaron-L", "name": { "family": "Nichols", "given": "Aaron L." }, "orcid": "0000-0001-9341-0049" }, { "id": "Bera-Kallol", "name": { "family": "Bera", "given": "Kallol" } }, { "id": "Bao-Huan", "name": { "family": "Bao", "given": "Huan" } }, { "id": "Bishara-Ishak", "name": { "family": "Bishara", "given": "Ishak" }, "orcid": "0000-0001-7563-1980" }, { "id": "Jeon-Janice-H", "name": { "family": "Jeon", "given": "Janice" }, "orcid": "0000-0002-8483-586X" }, { "id": "Mulcahy-Matthew-J", "name": { "family": "Mulcahy", "given": "Matthew J." }, "orcid": "0000-0001-5588-8762" }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce" } }, { "id": "O'Riordan-Saidhbhe-L", "name": { "family": "O'Riordan", "given": "Saidhbhe L." } }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Chapman-Edwin-R", "name": { "family": "Chapman", "given": "Edwin R." }, "orcid": "0000-0001-9787-8140" }, { "id": "Marvin-Jonathan-S", "name": { "family": "Marvin", "given": "Jonathan" }, "orcid": "0000-0003-2294-4515" }, { "id": "Looger-Loren-L", "name": { "family": "Looger", "given": "Loren" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Determining the pharmacokinetics of nicotinic drugs in the endoplasmic reticulum using biosensors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2019 Shivange et al. This article is distributed under the terms of an Attribution\u2013Noncommercial\u2013Share Alike\u2013No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution\u2013Noncommercial\u2013Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/). \n\nSubmitted: 1 August 2018; Revision received 5 November 2018; Accepted: 9 January 2019. Published February 4, 2019. \n\nHenry A. Lester and Amol V. Shivange thank the Janelia Research Campus for devoting hospitality and resources in the Visiting Scientist Program over a period of 3 y. We also thank Crystal N. Dilworth (early suggestion to use ProX-family PBPs), Jacob P. Keller (advice on pH and tubing), David P. Walton (analysis of N'MeNic), Michael R. Post (synthesis of N'MeNic), Peter H. Lee and Scott Sternson (purification of N'MeNic), Adela Nano and Jacqueline Barton (access to the ISS-K2 spectrofluorometer), Daniel Wagenaar (construction of LED light sources), Lauren M. Barnett (advice on photochemistry), Elizabeth K. Unger and Lin Tian (biosensors), Eric L. Snapp (ER imaging), Jennifer Lippincott-Schwartz (ER imaging), Eric R. Schreiter (biosensors), Tanner Lakin (cell culture), Kim Ruoho and Melissa Ramirez (AAV constructs), Baljit S. Khakh (advice), Mark Lobas (advice), Victoria J. Orphan and Fabai Wu (use and instruction on structured illumination microscope), Stephen Grant (assisted in confocal microscopy), Margaret Jefferies (excellent laboratory management at Janelia), and Purnima Deshpande (excellent laboratory management at the California Institute of Technology). \n\nThis research was supported by grants from US National Institutes of Health (DA036061, DA037161, DA043829, GM123582, GM007616, MH061876, NS097362, and NS034407), the California Tobacco-Related Disease Research Project (23XT-0007), the California Institute for Regenerative Medicine (EDUC2-08398), the Brain and Behavior Research Foundation (National Alliance for Research on Schizophrenia and Depression), the Howard Hughes Medical Institute, the Della Martin Foundation, Louis and Janet Fletcher, and California Institute of Technology SURF donors Paraskeva N. Danailov and Maria Chan. E.R. Chapman is an Investigator of the Howard Hughes Medical Institute. \n\nH.A. Lester previously had a consulting agreement with Pfizer, who manufacture varenicline. The authors declare no further competing financial interests. \n\nAuthor contributions: A.V. Shivange, P.M. Borden, A.K. Muthusamy, A.L. Nichols, K. Bera, H. Bao, I. Bishara, M.J. Mulcahy, B. Cohen, E.R. Chapman, J. Marvin, L. Looger, and H.A. Lester designed experiments. A.V. Shivange, P.M. Borden, A.K. Muthusamy, A.L. Nichols, K. Bera, H. Bao, I. Bishara, J. Jeon, M.J. Mulcahy, B. Cohen, S.L. O'Riordan, C. Kim and H.A. Lester performed experiments. J. Jeon performed simulations. A.V. Shivange, P.M. Borden, A.K. Muthusamy, J. Jeon, K. Bera, H. Bao, I. Bishara, M.J. Mulcahy, L. Looger, and H.A. Lester wrote the paper. A.V. Shivange, P.M. Borden, A.K. Muthusamy, A.L. Nichols, K. Bera, H. Bao, I. Bishara, J. Jeon, M.J. Mulcahy, B. Cohen, D.A. Dougherty, E.R. Chapman, L. Looger, and H.A. Lester revised the paper. \n\nMerritt C. Maduke served as editor.\n\nPublished - 738.full.pdf
Supplemental Material - JGP_201812201_DataS1.zip
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Supplemental Material - JGP_201812201_V9.mp4
Supplemental Material - JGP_201812201_sm.pdf
", "abstract": "Nicotine dependence is thought to arise in part because nicotine permeates into the endoplasmic reticulum (ER), where it binds to nicotinic receptors (nAChRs) and begins an \"inside-out\" pathway that leads to up-regulation of nAChRs on the plasma membrane. However, the dynamics of nicotine entry into the ER are unquantified. Here, we develop a family of genetically encoded fluorescent biosensors for nicotine, termed iNicSnFRs. The iNicSnFRs are fusions between two proteins: a circularly permutated GFP and a periplasmic choline-/betaine-binding protein engineered to bind nicotine. The biosensors iNicSnFR3a and iNicSnFR3b respond to nicotine by increasing fluorescence at [nicotine] <1 \u00b5M, the concentration in the plasma and cerebrospinal fluid of a smoker. We target iNicSnFR3 biosensors either to the plasma membrane or to the ER and measure nicotine kinetics in HeLa, SH-SY5Y, N2a, and HEK293 cell lines, as well as mouse hippocampal neurons and human stem cell\u2013derived dopaminergic neurons. In all cell types, we find that nicotine equilibrates in the ER within 10 s (possibly within 1 s) of extracellular application and leaves as rapidly after removal from the extracellular solution. The [nicotine] in the ER is within twofold of the extracellular value. We use these data to run combined pharmacokinetic and pharmacodynamic simulations of human smoking. In the ER, the inside-out pathway begins when nicotine becomes a stabilizing pharmacological chaperone for some nAChR subtypes, even at concentrations as low as \u223c10 nM. Such concentrations would persist during the 12 h of a typical smoker's day, continually activating the inside-out pathway by >75%. Reducing nicotine intake by 10-fold decreases activation to \u223c20%. iNicSnFR3a and iNicSnFR3b also sense the smoking cessation drug varenicline, revealing that varenicline also permeates into the ER within seconds. Our iNicSnFRs enable optical subcellular pharmacokinetics for nicotine and varenicline during an early event in the inside-out pathway.", "date": "2019-06", "date_type": "published", "publication": "Journal of General Physiology", "volume": "151", "number": "6", "publisher": "Rockefeller University Press", "pagerange": "738-757", "id_number": "CaltechAUTHORS:20190204-103658076", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190204-103658076", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA036061" }, { "agency": "NIH", "grant_number": "DA037161" }, { "agency": "NIH", "grant_number": "DA043829" }, { "agency": "NIH", "grant_number": "GM123582" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM007616" }, { "agency": "NIH", "grant_number": "MH061876" }, { "agency": "NIH", "grant_number": "NS097362" }, { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "23XT-0007" }, { "agency": "California Institute for Regenerative Medicine (CIRM)", "grant_number": "EDUC2-08398" }, { "agency": "Brain and Behavior Research Foundation" }, { "agency": "National Alliance for Research on Schizophrenia and Depression" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "Della Martin Foundation" }, { "agency": "Louis and Janet Fletcher" }, { "agency": "Caltech Summer Undergraduate Research Fellowship (SURF)" } ] }, "doi": "10.1085/jgp.201812201", "pmcid": "PMC6571994", "primary_object": { "basename": "JGP_201812201_sm.pdf", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_sm.pdf" }, "related_objects": [ { "basename": "738.full.pdf", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/738.full.pdf" }, { "basename": "JGP_201812201_DataS1.zip", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_DataS1.zip" }, { "basename": "JGP_201812201_V1.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V1.mp4" }, { "basename": "JGP_201812201_V7.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V7.mp4" }, { "basename": "JGP_201812201_V6.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V6.mp4" }, { "basename": "JGP_201812201_V8.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V8.mp4" }, { "basename": "JGP_201812201_V9.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V9.mp4" }, { "basename": "JGP_201812201_V2.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V2.mp4" }, { "basename": "JGP_201812201_V3.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V3.mp4" }, { "basename": "JGP_201812201_V4.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V4.mp4" }, { "basename": "JGP_201812201_V5.mp4", "url": "https://authors.library.caltech.edu/records/xmy9t-8yy14/files/JGP_201812201_V5.mp4" } ], "resource_type": "article", "pub_year": "2019", "author_list": "Shivange, Amol V.; Borden, Philip M.; et el." }, { "id": "https://authors.library.caltech.edu/records/h19d7-0bg42", "eprint_id": 92501, "eprint_status": "archive", "datestamp": "2023-08-22 01:17:49", "lastmod": "2023-10-20 15:46:39", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Bavan-Selvan", "name": { "family": "Bavan", "given": "Selvan" } }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "Henderson-Brandon-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Chronic menthol does not change stoichiometry or functional plasma membrane levels of mouse \u03b13\u03b24-containing nicotinic acetylcholine receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "Acetylcholine receptors; Addiction; Electrophysiology; Ion channels; Nicotinic cholinergic receptors; Patch clamp recording; Receptor trafficking", "note": "\u00a9 2019 The American Society for Pharmacology and Experimental Therapeutics. \n\nReceived October 5, 2018; accepted January 12, 2019; Published on January 22, 2019. \n\nWe are grateful to Suparna Patowary for performing preliminary experiments on chronic menthol treatment effects at mouse \u03b13\u03b24 nAChRs (Patowary et al., 2016). \n\nAuthor Contributions: Participated in research design: Bavan, Henderson, and Lester. Conducted experiments: Bavan and Kim. Contributed new reagents or analytic tools: not applicable. Performed data analysis: Bavan, Henderson, and Lester. Wrote or contributed to the writing of the manuscript: Bavan, Kim, Henderson, and Lester. \n\nThis study was supported by National Institutes of Health [Grants DA037743, DA046335, and DA036061].\n\nAccepted Version - mol.118.114769.full.pdf
", "abstract": "Heteromeric \u03b13\u03b24 nicotinic acetylcholine (ACh) receptors (nAChRs) are pentameric ligand-gated cation channels that include at least two \u03b13 and two \u03b24 subunits. They have functions in peripheral tissue and peripheral and central nervous systems. We examined the effects of chronic treatment with menthol, a major flavor additive in tobacco cigarettes and electronic nicotine delivery systems, on mouse \u03b13\u03b24 nAChRs transiently transfected into neuroblastoma-2a cells. Chronic menthol treatment at 500 nM, near the estimated menthol concentration in the brain following cigarette smoking, altered neither the [ACh]-response relationship nor Zn^(2+) sensitivity of ACh-evoked currents, suggesting that menthol does not change \u03b13\u03b24 nAChR subunit stoichiometry. Chronic menthol treatment failed to change the current density (peak current amplitude/cell capacitance) of 100 \u03bcM ACh-evoked currents. Chronic menthol treatment accelerated desensitization of 100 and 200 \u03bcM ACh-evoked currents. Chronic nicotine treatment (250 \u03bcM) decreased ACh-induced currents, and we found no additional effect of including chronic menthol. These data contrast with previously reported, marked effects of chronic menthol on \u03b22* nAChRs studied in the same expression system. Mechanistically, the data support the emerging interpretation that both chronic menthol and chronic nicotine act on nAChRs in the early exocytotic pathway, and that this pathway does not present a rate-limiting step to the export of \u03b13\u03b24 nAChRs; these nAChRs include endoplasmic reticulum (ER) export motifs but not ER retention motifs. Previous reports show that smoking mentholated cigarettes enhances tobacco addiction; but our results show that this effect is unlikely to arise via menthol actions on \u03b13\u03b24 nAChRs.", "date": "2019-04-01", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "95", "number": "4", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "398-407", "id_number": "CaltechAUTHORS:20190128-151017476", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190128-151017476", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA037743" }, { "agency": "NIH", "grant_number": "DA046335" }, { "agency": "NIH", "grant_number": "DA036061" } ] }, "doi": "10.1124/mol.118.114769", "pmcid": "PMC6399576", "primary_object": { "basename": "mol.118.114769.full.pdf", "url": "https://authors.library.caltech.edu/records/h19d7-0bg42/files/mol.118.114769.full.pdf" }, "resource_type": "article", "pub_year": "2019", "author_list": "Bavan, Selvan; Kim, Charlene H.; et el." }, { "id": "https://authors.library.caltech.edu/records/zfnag-vrw86", "eprint_id": 92410, "eprint_status": "archive", "datestamp": "2023-08-22 01:14:36", "lastmod": "2023-10-20 15:40:09", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Zhou-Chunyi", "name": { "family": "Zhou", "given": "Chunyi" } }, { "id": "Gu-Weixin", "name": { "family": "Gu", "given": "Weixin" } }, { "id": "Wu-Haichuan", "name": { "family": "Wu", "given": "Haichuan" } }, { "id": "Yan-Xiang", "name": { "family": "Yan", "given": "Xiang" } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Bidirectional dopamine modulation of excitatory and inhibitory synaptic inputs to subthalamic neuron subsets containing \u03b14\u03b22 or \u03b17 nAChRs", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Nicotinic acetylcholine receptor; \u03b14\u03b22; \u03b17; Subthalamic nucleus; Dopamine receptors", "note": "\u00a9 2019 Elsevier Ltd. \n\nReceived 10 August 2018, Revised 10 January 2019, Accepted 14 January 2019, Available online 17 January 2019. \n\nThis work was supported by Michael J. Fox Foundation (11345), National Institutes of Health (AG033954), the California Tobacco-Related Disease Research Program Grants, National Natural Science Foundation of China (81701100, 81870891), Fund for Jiangsu Province Specially-Appointed Professor (C.X., C.Z.), Natural Science Foundation of Jiangsu Province (BK20171160), The Natural Science Foundation of the Jiangsu Higher Education Institutions of China (17KJA320007, 18KJA320009), Jiangsu Province Fund for Dominant Discipline (Anesthesiology).", "abstract": "The subthalamic nucleus (STN) possesses microcircuits distinguished by subtypes of nicotinic acetylcholine receptors (nAChRs). Although dysfunction of the STN is well-known in Parkinson's disease, there is still little information about whether dopamine differentially modulates excitatory and inhibitory synaptic inputs to STN neurons expressing different nAChR subtypes. To address this issue, we performed brain slice patch-clamp recordings on STN neurons, while we pharmacologically manipulated dopaminergic inputs. In STN neuron subsets containing either \u03b14\u03b22 or \u03b17 nAChRs, D_1 and D_2 receptors respectively enhanced and inhibited spontaneous inhibitory and excitatory postsynaptic currents (sIPSCs and sEPSCs) and firing rates. The elevation of dopamine levels resulted in diverse regulations of synaptic transmission in these two neuron subsets, and interestingly, the dopamine regulation of sIPSCs significantly correlated with that of sEPSCs. Surprisingly, depletion of dopamine either by reserpine treatment or by unilateral 6-OHDA lesion of nigrostriatal dopaminergic neurons did not alter synaptic inputs to STN neurons, but STN neurons in the 6-OHDA-lesioned side exhibited hyperactivity. In summary, dopamine regulated both GABAergic and glutamatergic synaptic inputs to STN neuron subsets containing either \u03b14\u03b22 or \u03b17 nAChRs, forming a balancing machinery to control neuronal activity. In parkinsonian mice, postsynaptic mechanisms may exist and contribute to the hyperactivity of STN neurons.", "date": "2019-04", "date_type": "published", "publication": "Neuropharmacology", "volume": "148", "publisher": "Elsevier", "pagerange": "220-228", "id_number": "CaltechAUTHORS:20190123-075807331", "issn": "0028-3908", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190123-075807331", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Michael J. Fox Foundation", "grant_number": "11345" }, { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "National Natural Science Foundation of China", "grant_number": "81701100" }, { "agency": "National Natural Science Foundation of China", "grant_number": "81870891" }, { "agency": "Fund for Jiangsu Province" }, { "agency": "Natural Science Foundation of Jiangsu Province", "grant_number": "BK20171160" }, { "agency": "Jiangsu Higher Education Institutions of China", "grant_number": "17KJA320007" }, { "agency": "Jiangsu Higher Education Institutions of China", "grant_number": "18KJA320009" }, { "agency": "Jiangsu Province Fund for Dominant Discipline" } ] }, "doi": "10.1016/j.neuropharm.2019.01.015", "resource_type": "article", "pub_year": "2019", "author_list": "Zhou, Chunyi; Gu, Weixin; et el." }, { "id": "https://authors.library.caltech.edu/records/1ygsc-yjn19", "eprint_id": 92213, "eprint_status": "archive", "datestamp": "2023-08-22 00:15:13", "lastmod": "2023-10-20 00:05:26", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Henderson-Brandon-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Grant-Stephen", "name": { "family": "Grant", "given": "Stephen" }, "orcid": "0000-0003-0923-8886" }, { "id": "Chu-Betty-W", "name": { "family": "Chu", "given": "Betty W." } }, { "id": "Shahoei-Rezvan", "name": { "family": "Shahoei", "given": "Rezvan" } }, { "id": "Huard-Stephanie-M", "name": { "family": "Huard", "given": "Stephanie M." }, "orcid": "0000-0002-2333-1852" }, { "id": "Saladi-Shyam-M", "name": { "family": "Saladi", "given": "Shyam S. M." }, "orcid": "0000-0001-9701-3059" }, { "id": "Tajkhorshid-Emad", "name": { "family": "Tajkhorshid", "given": "Emad" }, "orcid": "0000-0001-8434-1010" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Menthol stereoisomers exhibit different effects on \u03b14\u03b22 nAChR upregulation and dopamine neuron spontaneous firing", "ispublished": "pub", "full_text_status": "public", "keywords": "cigarettes; electronic nicotine delivery systems; nicotine; nicotine addiction; Xenopus oocyte", "note": "\u00a9 2019 by the Society for Neuroscience. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. \n\nThe authors declare no conflict of interest. \n\nFunding was provided by National Institutes of Health (NIH) DA036061, DA037161, and DA037743 (HAL); NIH DA033721 and DA040047 (BJH); and Marshall University Research Corporation (BJH); NIH GM104601, NIH GM087519, and NIH GM067887 (RS and ET). Computing resources were provided by Blue Waters at National Center for Supercomputing Applications and Extreme Science and Engineering Discovery Environment (XSEDE) (Grant TG-MCA06N060 to ET). \n\nB.J.H, S.G and B.W.C Designates co-first authorship.\n\nPublished - ENEURO.0465-18.2018.pdf
", "abstract": "Menthol contributes to poor cessation rates among smokers, in part because menthol enhances nicotine reward and reinforcement. Mentholated tobacco products contain (\u2212)-menthol and (+)-menthol, in varying proportions. We examined these two menthol stereoisomers for their ability to upregulate \u03b14\u03b22 nAChRs and to alter dopamine neuron firing frequency using long-term, low-dose (\u2264 500 nM) exposure that is pharmacologically relevant to smoking. We found that (\u2212)-menthol upregulates \u03b14\u03b22 nAChRs while (+)-menthol does not. We also found that (\u2212)-menthol decreases dopamine neuron baseline firing and dopamine neuron excitability, while (+)-menthol exhibits no effect. We then examined both stereoisomers for their ability to inhibit \u03b14\u03b22 nAChR function at higher concentrations (>10 \u00b5M) using the Xenopus oocyte expression system. To probe for the potential binding site of menthol, we conducted flooding simulations and site-directed mutagenesis. We found that menthol likely binds to the 9' position on the TM2 helix. We found that menthol inhibition is dependent on the end-to-end distance of the side chain at the 9' residue. Additionally, we have found that (\u2212)-menthol is only modestly (\u223c25%) more potent than (+)-menthol at inhibiting wildtype \u03b14\u03b22 nAChRs and a series of L9' mutant nAChRs. These data reveal that menthol exhibits a stereoselective effect on nAChRs and that the stereochemical effect is much greater for long-term, sub \u00b5M exposure in mice than for acute, higher level exposure. We hypothesize that of the two menthol stereoisomers, only (\u2212)-menthol plays a role in enhancing nicotine reward through nAChRs on dopamine neurons.", "date": "2018-11", "date_type": "published", "publication": "eNeuro", "volume": "5", "number": "6", "publisher": "Society for Neuroscience", "pagerange": "Art. No. e0465-18.2018", "id_number": "CaltechAUTHORS:20190111-101254546", "issn": "2373-2822", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190111-101254546", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA036061" }, { "agency": "NIH", "grant_number": "DA037161" }, { "agency": "NIH", "grant_number": "DA037743" }, { "agency": "NIH", "grant_number": "DA033721" }, { "agency": "NIH", "grant_number": "DA040047" }, { "agency": "NIH", "grant_number": "GM104601" }, { "agency": "NIH", "grant_number": "GM087519" }, { "agency": "NIH", "grant_number": "GM067887" } ] }, "doi": "10.1523/eneuro.0465-18.2018", "pmcid": "PMC6325563", "primary_object": { "basename": "ENEURO.0465-18.2018.pdf", "url": "https://authors.library.caltech.edu/records/1ygsc-yjn19/files/ENEURO.0465-18.2018.pdf" }, "resource_type": "article", "pub_year": "2018", "author_list": "Henderson, Brandon J.; Grant, Stephen; et el." }, { "id": "https://authors.library.caltech.edu/records/0zgtw-kvm05", "eprint_id": 89894, "eprint_status": "archive", "datestamp": "2023-08-19 11:33:24", "lastmod": "2023-10-18 23:04:33", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nichols-Aaron-L", "name": { "family": "Nichols", "given": "Aaron L." }, "orcid": "0000-0001-9341-0049" }, { "id": "Noridomi-Kaori", "name": { "family": "Noridomi", "given": "Kaori" } }, { "id": "Hughes-Christopher-R", "name": { "family": "Hughes", "given": "Christopher R." } }, { "id": "Jalali-Yazdi-Farzad", "name": { "family": "Jalali-Yazdi", "given": "Farzad" } }, { "id": "Eaton-J-Brek", "name": { "family": "Eaton", "given": "J. Brek" } }, { "id": "Lai-Lan-Huong", "name": { "family": "Lai", "given": "Lan Huong" } }, { "id": "Advani-Gaurav", "name": { "family": "Advani", "given": "Gaurav" } }, { "id": "Lukas-Ronald-J", "name": { "family": "Lukas", "given": "Ronald J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Chen-Lin", "name": { "family": "Chen", "given": "Lin" } }, { "id": "Roberts-Richard-W", "name": { "family": "Roberts", "given": "Richard W." }, "orcid": "0000-0002-8587-5097" } ] }, "title": "\u03b11-FANGs: Protein Ligands Selective for the \u03b1-Bungarotoxin Site of the \u03b11-Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2018 American Chemical Society. \n\nReceived: June 1, 2018; Accepted: July 30, 2018; Published: July 30, 2018. \n\nWe would like to thank Caitlin Nichols for her acronym expertise and Dr. Matthew Mulcahy for his digital prowess. \n\nSupported by National Institutes of Health Grants R01 AI085583 and R01 DA037161. \n\nThe authors declare no competing financial interest.\n\nAccepted Version - nihms-1754167.pdf
Supplemental Material - cb8b00513_si_002.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are pentameric ligand-gated ion channels that play a central role in neuronal and neuromuscular signal transduction. Here, we have developed FANG ligands, fibronectin antibody-mimetic nicotinic acetylcholine receptor-generated ligands, using mRNA display. We generated a 1 trillion-member primary e10FnIII library to target a stabilized \u03b11 nicotinic subunit (\u03b1211). This library yielded 270000 independent potential protein binding ligands. The lead sequence, \u03b11-FANG1, represented 25% of all library sequences, showed the highest-affinity binding, and competed with \u03b1-bungarotoxin (\u03b1-Btx). To improve this clone, a new library based on \u03b11-FANG1 was subjected to heat, protease, binding, off-rate selective pressures, and point mutations. This resulted in \u03b11-FANG2 and \u03b11-FANG3. These proteins bind \u03b1211 with KDvalues of 3.5 nM and 670 pM, respectively, compete with \u03b1-Btx, and show improved subunit specificity. \u03b11-FANG3 is thermostable (T_m = 62 \u00b0C) with a 6 kcal/mol improvement in folding free energy compared with that of the parent \u03b11-FANG1. \u03b11-FANG3 competes directly with the \u03b1-Btx binding site of intact neuromuscular heteropentamers [(\u03b11)_2\u03b21\u03b3\u03b4] in mammalian culture-derived cellular membranes and in Xenopus laevis oocytes expressing these nAChRs. This work demonstrates that mRNA display against a monomeric ecto-domain of a pentamer has the capability to select ligands that bind that subunit in both a monomeric and a pentameric context. Overall, our work provides a route to creating a new family of stable, well-behaved proteins that specifically target this important receptor family.", "date": "2018-09-21", "date_type": "published", "publication": "ACS Chemical Biology", "volume": "13", "number": "9", "publisher": "American Chemical Society", "pagerange": "2568-2576", "id_number": "CaltechAUTHORS:20180924-141114616", "issn": "1554-8929", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180924-141114616", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "R01 AI085583" }, { "agency": "NIH", "grant_number": "R01 DA037161" } ] }, "doi": "10.1021/acschembio.8b00513", "pmcid": "PMC8763392", "primary_object": { "basename": "cb8b00513_si_002.pdf", "url": "https://authors.library.caltech.edu/records/0zgtw-kvm05/files/cb8b00513_si_002.pdf" }, "related_objects": [ { "basename": "nihms-1754167.pdf", "url": "https://authors.library.caltech.edu/records/0zgtw-kvm05/files/nihms-1754167.pdf" } ], "resource_type": "article", "pub_year": "2018", "author_list": "Nichols, Aaron L.; Noridomi, Kaori; et el." }, { "id": "https://authors.library.caltech.edu/records/wapxg-wd930", "eprint_id": 86961, "eprint_status": "archive", "datestamp": "2023-08-21 23:53:02", "lastmod": "2023-10-18 20:44:23", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Subramaniam-S-R", "name": { "family": "Subramaniam", "given": "Sudhakar R." } }, { "id": "Magen-I", "name": { "family": "Magen", "given": "Iddo" } }, { "id": "Bove-N", "name": { "family": "Bove", "given": "Nicholas" } }, { "id": "Zhu-Chunni", "name": { "family": "Zhu", "given": "Chunni" } }, { "id": "Lemesre-V", "name": { "family": "Lemesre", "given": "Vincent" } }, { "id": "Dutta-G", "name": { "family": "Dutta", "given": "Garima" } }, { "id": "Elias-C-J", "name": { "family": "Elias", "given": "Chris Jean" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Chesselet-M-F", "name": { "family": "Chesselet", "given": "Marie-Francoise" } } ] }, "title": "Chronic nicotine improves cognitive and social impairment in mice overexpressing wild type \u03b1-synuclein", "ispublished": "pub", "full_text_status": "public", "keywords": "Parkinson's disease; Mouse model; \u03b1-synuclein; Nicotine; Motor deficits; Cognitive deficits; Social impairment", "note": "\u00a9 2018 Elsevier Inc. \n\nReceived 19 March 2018, Revised 7 May 2018, Accepted 29 May 2018, Available online 1 June 2018. \n\nThis work was supported by the Michael J. Fox Foundation (MJFF) Target Validation, 2011, by the Caltech-UCLA Joint Center for Translational Medicine (JCTM, UCLA-CALTECH-77857), by NIH grant AG-033954, by gifts to the Center for the Study of Parkinson's disease at UCLA, and by gifts from Louis and Janet Fletcher at Caltech. We thank Dr. Franziska Richter and Sheri McKinney for help in designing the study. We also thank undergraduate students Bansi Patel, Jacky Kwong, Sean Campeau and Diana Dinh for their help with behavior rating and histology. MFC has received honoraria (for service as a reviewer) and travel reimbursement from MJFF. \n\nThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.\n\nAccepted Version - nihms-976305.pdf
", "abstract": "In addition to dopaminergic and motor deficits, patients with Parkinson's disease (PD) suffer from non-motor symptoms, including early cognitive and social impairment, that do not respond well to dopaminergic therapy. Cholinergic deficits may contribute to these problems, but cholinesterase inhibitors have limited efficacy. Mice over-expressing \u03b1-synuclein, a protein critically associated with PD, show deficits in cognitive and social interaction tests, as well as a decrease in cortical acetylcholine. We have evaluated the effects of chronic administration of nicotine in mice over-expressing wild type human \u03b1-synuclein under the Thy1-promoter (Thy1-aSyn mice). Nicotine was administered subcutaneously by osmotic minipump for 6\u202fmonths from 2 to 8\u202fmonths of age at 0.4\u202fmg/kg/h and 2.0\u202fmg/kg/h. The higher dose was toxic in the Thy1-aSyn mice, but the low dose was well tolerated and both doses ameliorated cognitive impairment in Y-maze performance after 5\u202fmonths of treatment. In a separate cohort of Thy1-aSyn mice, nicotine was administered at the lower dose for one month beginning at 5\u202fmonths of age. This treatment partially eliminated the cognitive deficit in novel object recognition and social impairment. In contrast, chronic nicotine did not improve motor deficits after 2, 4 or 6\u202fmonths of treatment, nor modified \u03b1-synuclein aggregation, tyrosine hydroxylase immunostaining, synaptic and dendritic markers, or microglial activation in Thy1-aSyn mice. These results suggest that cognitive and social impairment in synucleinopathies like PD may result from deficits in cholinergic neurotransmission and may benefit from chronic administration of nicotinic agonists.", "date": "2018-09", "date_type": "published", "publication": "Neurobiology of Disease", "volume": "117", "publisher": "Elsevier", "pagerange": "170-180", "id_number": "CaltechAUTHORS:20180611-095839803", "issn": "0969-9961", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180611-095839803", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Michael J. Fox Foundation" }, { "agency": "Caltech-UCLA Joint Center for Translational Medicine" }, { "agency": "NIH", "grant_number": "AG-033954" } ] }, "doi": "10.1016/j.nbd.2018.05.018", "pmcid": "PMC6051902", "primary_object": { "basename": "nihms-976305.pdf", "url": "https://authors.library.caltech.edu/records/wapxg-wd930/files/nihms-976305.pdf" }, "resource_type": "article", "pub_year": "2018", "author_list": "Subramaniam, Sudhakar R.; Magen, Iddo; et el." }, { "id": "https://authors.library.caltech.edu/records/r6e1d-xzq14", "eprint_id": 85557, "eprint_status": "archive", "datestamp": "2023-08-19 08:58:10", "lastmod": "2023-10-20 21:54:56", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Banala-S", "name": { "family": "Banala", "given": "Sambashiva" } }, { "id": "Arvin-M-C", "name": { "family": "Arvin", "given": "Matthew C." }, "orcid": "0000-0002-1744-2142" }, { "id": "Bannon-N-M", "name": { "family": "Bannon", "given": "Nicholas M." } }, { "id": "Jin-Xiao-Tao", "name": { "family": "Jin", "given": "Xiao-Tao" } }, { "id": "Macklin-J-J", "name": { "family": "Macklin", "given": "John J." } }, { "id": "Wang-Yong", "name": { "family": "Wang", "given": "Yong" }, "orcid": "0000-0001-7547-1542" }, { "id": "Peng-Can", "name": { "family": "Peng", "given": "Can" } }, { "id": "Zhao-Guiqing", "name": { "family": "Zhao", "given": "Guiqing" } }, { "id": "Marshall-J-J", "name": { "family": "Marshall", "given": "John J." } }, { "id": "Gee-K-R", "name": { "family": "Gee", "given": "Kyle R." } }, { "id": "Wokosin-D-L", "name": { "family": "Wokosin", "given": "David L." } }, { "id": "Kim-V-J", "name": { "family": "Kim", "given": "Veronica J." } }, { "id": "McIntosh-J-M", "name": { "family": "McIntosh", "given": "J. Michael" } }, { "id": "Contractor-A", "name": { "family": "Contractor", "given": "Anis" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Kozorovitskiy-Y", "name": { "family": "Kozorovitskiy", "given": "Yevgenia" }, "orcid": "0000-0002-3710-1484" }, { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Lavis-L-D", "name": { "family": "Lavis", "given": "Luke D." }, "orcid": "0000-0002-0789-6343" } ] }, "title": "Photoactivatable drugs for nicotinic optopharmacology", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2018 Macmillan Publishers Limited, part of Springer Nature. \n\nReceived: 26 September 2017; Accepted: 26 February 2018; Published: 26 March 2018. \n\nData availability. The data that support the findings of this study, if not explicitly contained within the text, supplementary figures, or Supplementary Note 1, are available from the corresponding authors upon reasonable request. Source data for Figures 1 and 2 and Supplementary Figures 1\u20136 are available online. \n\nWe thank members of the Drenan and Lavis laboratories for helpful advice and discussion, and T. Lerner (Northwestern University, Chicago, Illinois, USA) for contributing viral reagents. This work was supported by the Howard Hughes Medical Institute (to S.B., J.J.M., and L.D.L.), the US National Institutes of Health (NIH) (grants DA035942 and DA040626 to R.M.D., MH099114 to A.C., DA037161 to H.A.L., NS054850 to D.J. Surmeier, and GM103801 and GM48677 to J.M.M.), the PhRMA Foundation (fellowship to M.C.A.), the Arnold and Mabel Beckman Foundation (Beckman Young Investigator Award to Y.K.), the Bernice E. Bumpus Foundation (Early Career Innovation Award to Y.K.), the Rita Allen Foundation (to Y.K.), the Searle Scholars Program (to Y.K.), the Alfred P. Sloan Foundation (Sloan Research Fellowship to Y.K.), NINDS (grant NINDS F32 NS103243 to N.M.B.), the JPB Foundation, and Northwestern University. \n\nAuthor Contributions: R.M.D., M.C.A., H.A.L., S.B., K.R.G., and L.D.L. conceived the project. M.C.A., N.M.B., D.L.W., X.-T.J., J.J.M., Y.W., C.P., G.Z., V.J.K., J.J.M., A.C., Y.K., R.M.D., S.B., and L.D.L. planned and/or executed experiments. D.L.W., Y.K., J.M.M., and K.R.G. contributed essential reagents and expertise. R.M.D., M.C.A., S.B., and L.D.L. wrote the paper with input from all other authors. R.M.D. and L.D.L. supervised all aspects of the work. \n\nCompeting interests: K.R.G. is an employee of Thermo Fisher Scientific and has stock options. All other authors declare no competing interests.\n\nAccepted Version - nihms947829.pdf
Submitted - 260232.full.pdf
Supplemental Material - nmeth.4637-S1.pdf
Supplemental Material - nmeth.4637-S2.pdf
", "abstract": "Photoactivatable pharmacological agents have revolutionized neuroscience, but the palette of available compounds is limited. We describe a general method for caging tertiary amines by using a stable quaternary ammonium linkage that elicits a red shift in the activation wavelength. We prepared a photoactivatable nicotine (PA-Nic), uncageable via one- or two-photon excitation, that is useful to study nicotinic acetylcholine receptors (nAChRs) in different experimental preparations and spatiotemporal scales.", "date": "2018-05", "date_type": "published", "publication": "Nature Methods", "volume": "15", "number": "5", "publisher": "Nature Publishing Group", "pagerange": "347-350", "id_number": "CaltechAUTHORS:20180402-095419703", "issn": "1548-7091", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180402-095419703", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "NIH", "grant_number": "DA035942" }, { "agency": "NIH", "grant_number": "DA040626" }, { "agency": "NIH", "grant_number": "MH099114" }, { "agency": "NIH", "grant_number": "DA037161" }, { "agency": "NIH", "grant_number": "NS054850" }, { "agency": "NIH", "grant_number": "GM103801" }, { "agency": "NIH", "grant_number": "GM48677" }, { "agency": "PhRMA Foundation" }, { "agency": "Arnold and Mabel Beckman Foundation" }, { "agency": "Bernice E. Bumpus Foundation" }, { "agency": "Rita Allen Foundation" }, { "agency": "Searle Scholars Program" }, { "agency": "Alfred P. Sloan Foundation" }, { "agency": "NIH", "grant_number": "F32 NS103243" }, { "agency": "JPB Foundation" }, { "agency": "Northwestern University" } ] }, "doi": "10.1038/nmeth.4637", "pmcid": "PMC5923430", "primary_object": { "basename": "260232.full.pdf", "url": "https://authors.library.caltech.edu/records/r6e1d-xzq14/files/260232.full.pdf" }, "related_objects": [ { "basename": "nihms947829.pdf", "url": "https://authors.library.caltech.edu/records/r6e1d-xzq14/files/nihms947829.pdf" }, { "basename": "nmeth.4637-S1.pdf", "url": "https://authors.library.caltech.edu/records/r6e1d-xzq14/files/nmeth.4637-S1.pdf" }, { "basename": "nmeth.4637-S2.pdf", "url": "https://authors.library.caltech.edu/records/r6e1d-xzq14/files/nmeth.4637-S2.pdf" } ], "resource_type": "article", "pub_year": "2018", "author_list": "Banala, Sambashiva; Arvin, Matthew C.; et el." }, { "id": "https://authors.library.caltech.edu/records/5xb62-4ms53", "eprint_id": 86574, "eprint_status": "archive", "datestamp": "2023-08-19 07:43:54", "lastmod": "2023-10-18 19:44:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Bavan-S", "name": { "family": "Bavan", "given": "Selvan" } }, { "id": "Patowary-S", "name": { "family": "Patowary", "given": "Suparna" } }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Effects of Chronic Menthol at Alpha3Beta4 (\u03b13\u03b24)-Containing Nicotinic Acetylcholine Receptors", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2018 Biophysical Society. \n\nAvailable online 6 February 2018.", "abstract": "Some heteropentameric nicotinic acetylcholine receptors (nAChRs) are up-regulated by chronic nicotine. Menthol is present in \u223c 30% of tobacco cigarettes sold in the United States. Compared with non-mentholated cigarettes, menthol-containing cigarettes are associated with reduced smoking cessation. Chronic menthol favors the lower sensitivity (\u03b14)3(\u03b22)2, whereas chronic nicotine favors the higher sensitivity (\u03b14)2(\u03b22)3 stoichiometry. Following chronic nicotine treatment, total cell lysates displayed a shift in stoichiometry towards (\u03b13)2(\u03b24)3 from (\u03b13)3(\u03b24)2. \u03b13\u03b24 nAChRs are highly expressed in the medial habenula and interpeduncular nuclei, which are involved in reward processing, and possibly nicotine addiction and withdrawal. We studied effects following chronic treatment (at least 24 hours) of 500 nM menthol at \u03b13\u03b24 nAChR using F\u00f6rster resonance energy transfer (FRET), total internal reflection fluorescence microscopy (TIRFM), and whole-cell patch-clamp electrophysiology of Neuro-2a cells transiently expressing fluorescently labeled subunits. FRET experiments indicated a shift in stoichiometry toward (\u03b13)3(\u03b24)2 from (\u03b13)2(\u03b24)3. TIRFM experiments revealed \u03b13 subunit up-regulation in the endoplasmic reticulum and \u03b13\u03b24 nAChR reduction in the plasma membrane. Our FRET experiments, however, include contributions from intracellular \u03b13\u03b24 nAChR stoichiometry. In contrast, patch clamp experiments measuring Zn2+ inhibition of acetylcholine-evoked currents indicate exclusively the functional cell surface \u03b13\u03b24 nAChR stoichiometry. Neither chronic menthol, chronic nicotine, nor combined chronic menthol and nicotine detectably alters Zn2+ inhibition of acetylcholine, showing that neither alters functional plasma membrane \u03b13\u03b24 nAChR stoichiometry. Furthermore, chronic menthol treatment shifts by < 1.5-fold the EC50 of acetylcholine at \u03b13\u03b24 nAChRs. These findings are consistent with our fluorescence-based experiments showing a reduction in endoplasmic reticulum exit sites following chronic menthol, which consequently reduces \u03b13\u03b24 nAChR delivery to the plasma membrane. Therefore, despite their intracellular effects, neither menthol nor nicotine influences cell surface \u03b13\u03b24 nAChR stoichiometry. Support: DA037743, DA036061, DA40047.", "date": "2018-02-02", "date_type": "published", "publication": "Biophysical Journal", "volume": "114", "number": "3", "publisher": "Biophysical Society", "pagerange": "296A", "id_number": "CaltechAUTHORS:20180523-144402903", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180523-144402903", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA037743" }, { "agency": "NIH", "grant_number": "DA036061" }, { "agency": "NIH", "grant_number": "DA40047" } ] }, "doi": "10.1016/j.bpj.2017.11.1691", "resource_type": "article", "pub_year": "2018", "author_list": "Bavan, Selvan; Patowary, Suparna; et el." }, { "id": "https://authors.library.caltech.edu/records/5vgys-d3273", "eprint_id": 86577, "eprint_status": "archive", "datestamp": "2023-08-19 07:44:16", "lastmod": "2023-10-18 19:44:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Muthusamy-A-K", "name": { "family": "Muthusamy", "given": "Anand K." }, "orcid": "0000-0003-1041-914X" }, { "id": "Shivange-A-V", "name": { "family": "Shivange", "given": "Amol V." } }, { "id": "Nichols-A-L", "name": { "family": "Nichols", "given": "Aaron L." }, "orcid": "0000-0001-9341-0049" }, { "id": "Kamajaya-A", "name": { "family": "Kamajaya", "given": "Aron" } }, { "id": "Jeon-Janice-H", "name": { "family": "Jeon", "given": "Janice" }, "orcid": "0000-0002-8483-586X" }, { "id": "Borden-P-M", "name": { "family": "Borden", "given": "Philip M." } }, { "id": "Marvin-J-S", "name": { "family": "Marvin", "given": "Jonathan S." }, "orcid": "0000-0003-2294-4515" }, { "id": "Unger-E-K", "name": { "family": "Unger", "given": "Elizabeth K." } }, { "id": "Bao-Huan", "name": { "family": "Bao", "given": "Huan" } }, { "id": "Chapman-E-R", "name": { "family": "Chapman", "given": "Edwin R." }, "orcid": "0000-0001-9787-8140" }, { "id": "Tian-Lin", "name": { "family": "Tian", "given": "Lin" }, "orcid": "0000-0001-7012-6926" }, { "id": "Looger-L-L", "name": { "family": "Looger", "given": "Loren L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Microscopy Using Fluorescent Drug Biosensors for \"Inside-Out Pharmacology\"", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2018 Biophysical Society. \n\nAvailable online 6 February 2018.", "abstract": "Neuropharmacology offers many tools for connecting molecular to acute behavioral phenomena but has few tools to explain effects of chronic drugs. In the biophysically based \"inside-out\" approach to neuropharmacology, drugs bind to their nascent targets within the endoplasmic reticulum (ER) (i.e. pharmacological chaperoning). Now we visualize, quantify, and time the first steps: drugs entering the cell and entering organelles.\n\nOur fluorescent biosensors are built on an OpuBC-GFP fusion scaffold. OpuBC is a bacterial periplasmic binding protein with two key features: a cation-\u03c0 box, favorable for binding amines well represented in drugs, and a binding-induced \"Venus fly-trap\" conformational change. The suitably mutated OpuBC domain is connected, at the hinge regions, to a circularly permuted \"superfolder\" GFP (cpGFP). Aided by structural information, we use directed evolution to create a family of drug-sensing biosensors meeting the criterion of \u0394F/F0 > 1 at 1 \u03bcM drug.\n\nOur proof of principle study concerns nicotine entering the ER, as measured by an \"intensity-based nicotine-sensing fluorescent reporter\" (iNicSnFR). Previous data demonstrated that exposure to nicotine causes changes in number and stoichiometry of nicotinic receptors by chaperoning within the ER; however nicotine itself entering the ER had not yet been measured. In live cell imaging of an iNicSnFR targeted to the ER, we found that nicotine enters the ER within 10 s of application at concentrations experienced by a cigarette smoker. Moreover, we found that varenicline, a smoking cessation drug, enters the ER almost as rapidly as nicotine, helping to explain varenicline's biochemical and behavioral effects. \n\nWe are currently developing other \"iDrugSnFRs\" for antidepressants, antipsychotics, and opioids. These tools to study subcellular pharmacokinetics will help to clarify chronic effects of several families of neural drugs. \n\nSupport: DA037161, GM123582, NARSAD, California TRDRP, and HHMI.", "date": "2018-02-02", "date_type": "published", "publication": "Biophysical Journal", "volume": "114", "number": "3", "publisher": "Biophysical Society", "pagerange": "358A", "id_number": "CaltechAUTHORS:20180523-150441864", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180523-150441864", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA037161" }, { "agency": "NIH", "grant_number": "GM123582" }, { "agency": "National Alliance for Research on Schizophrenia and Depression (NARSAD)" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Howard Hughes Medical Institute (HHMI)" } ] }, "doi": "10.1016/j.bpj.2017.11.1990", "resource_type": "article", "pub_year": "2018", "author_list": "Muthusamy, Anand K.; Shivange, Amol V.; et el." }, { "id": "https://authors.library.caltech.edu/records/7b2js-f0v95", "eprint_id": 83816, "eprint_status": "archive", "datestamp": "2023-08-19 06:30:50", "lastmod": "2023-10-17 23:32:17", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Parker-R-L", "name": { "family": "Parker", "given": "Rell L." } }, { "id": "O'Neill-H-C", "name": { "family": "O'Neill", "given": "Heidi C." } }, { "id": "Henley-B-M", "name": { "family": "Henley", "given": "Beverley M." } }, { "id": "Wageman-C-R", "name": { "family": "Wageman", "given": "Charles R." } }, { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Miwa-J-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Deletion of lynx1 reduces the function of \u03b16* nicotinic receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2017 Parker et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. \n\nReceived: June 14, 2017; Accepted: November 13, 2017; Published: December 5, 2017. \n\nData Availability Statement: All relevant data are within the paper. \n\nThis research was supported by funds provided by California Tobacco-Related Diseases Research Program (http://www.trdrp.org), Grant 22DT-0008 to RLP, and 19KT-0032 to JMM. Additional support was provided by NIH / NIDA (https://www.drugabuse.gov/) grants, DA003194, DA012242, and P30-DA015663 to MJM, DA017279 to HAL, and DA019375 to HAL and MJM, DA030396 and DA035942 to RMD, and DA033831, DA032464 to JMM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. \n\nThe authors have declared that no competing interests exist. \n\nWe thank J. Michael McIntosh (University of Utah, Salt Lake City, Utah) for providing \u03b1-CtxMII. WE thank Kenneth J. Kellar (Georgetown University, Washington DC) for providing 6-I-epibatidine. We thank Xiomara Perez (Center for Health Sciences, SRI International, Menlo Park, CA) for help with electrochemistry, Andrew Steele (Department of Biological Sciences, California State Polytechnic University, Pomona, CA) for help with automated behavior analyses, and Sreelaxmi Varkala for help with other behavioral analyses. \n\nAuthor Contributions: Conceptualization: Rell L. Parker, Ryan M. Drenan, Michael J. Marks, Julie M. Miwa, Sharon R. Grady, Henry A. Lester. Data curation: Rell L. Parker, Michael J. Marks, Sharon R. Grady. Formal analysis: Rell L. Parker, Heidi C. O'Neill, Beverley M. Henley, Michael J. Marks, Sharon R. Grady, Henry A. Lester. Funding acquisition: Rell L. Parker, Ryan M. Drenan, Julie M. Miwa.\nInvestigation: Rell L. Parker, Heidi C. O'Neill, Beverley M. Henley, Charles R. Wageman, Ryan M. Drenan, Michael J. Marks, Julie M. Miwa, Sharon R. Grady. Methodology: Rell L. Parker, Heidi C. O'Neill, Ryan M. Drenan, Michael J. Marks, Julie M. Miwa, Sharon R. Grady, Henry A. Lester. Project administration: Henry A. Lester. Resources: Ryan M. Drenan, Henry A. Lester. Supervision: Henry A. Lester. Visualization: Henry A. Lester. Writing \u00b1 original draft: Rell L. Parker, Heidi C. O'Neill, Beverley M. Henley, Ryan M. Drenan, Michael J. Marks, Julie M. Miwa, Sharon R. Grady, Henry A. Lester. Writing \u00b1 review & editing: Rell L. Parker, Heidi C. O'Neill, Ryan M. Drenan, Michael J. Marks, Julie M. Miwa, Sharon R. Grady, Henry A. Lester.\n\nPublished - journal.pone.0188715.pdf
", "abstract": "The \u03b16 nicotinic acetylcholine receptor (nAChR) subunit is an attractive drug target for treating nicotine addiction because it is present at limited sites in the brain including the reward pathway. Lynx1 modulates several nAChR subtypes; lynx1-nAChR interaction sites could possibly provide drug targets. We found that dopaminergic cells from the substantia nigra pars compacta (SNc) express lynx1 mRNA transcripts and, as assessed by co-immunoprecipitation, \u03b16 receptors form stable complexes with lynx1 protein, although co-transfection with lynx1 did not affect nicotine-induced currents from cell lines transfected with \u03b16 and \u03b22. To test whether lynx1 is important for the function of \u03b16 nAChRs in vivo, we bred transgenic mice carrying a hypersensitive mutation in the \u03b16 nAChR subunit (\u03b16L9\u2032S) with lynx1 knockout mice, providing a selective probe of the effects of lynx1 on \u03b16* nAChRs. Lynx1 removal reduced the \u03b16 component of nicotine-mediated rubidium efflux and dopamine (DA) release from synaptosomal preparations with no effect on numbers of \u03b16\u03b22 binding sites, indicating that lynx1 is functionally important for \u03b16* nAChR activity. No effects of lynx1 removal were detected on nicotine-induced currents in slices from SNc, suggesting that lynx1 affects presynaptic \u03b16* nAChR function more than somatic function. In the absence of agonist, lynx1 removal did not alter DA release in dorsal striatum as measured by fast scan cyclic voltammetry. Lynx1 removal affected some behaviors, including a novel-environment assay and nicotine-stimulated locomotion. Trends in 24-hour home-cage behavior were also suggestive of an effect of lynx1 removal. Conditioned place preference for nicotine was not affected by lynx1 removal. The results show that some functional and behavioral aspects of \u03b16-nAChRs are modulated by lynx1.", "date": "2017-12-05", "date_type": "published", "publication": "PLoS ONE", "volume": "12", "number": "12", "publisher": "Public Library of Science", "pagerange": "Art. No. e0188715", "id_number": "CaltechAUTHORS:20171211-155148787", "issn": "1932-6203", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20171211-155148787", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "22DT-0008" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19KT-0032" }, { "agency": "NIH", "grant_number": "DA003194" }, { "agency": "NIH", "grant_number": "DA012242" }, { "agency": "NIH", "grant_number": "P30-DA015663" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "DA019375" }, { "agency": "NIH", "grant_number": "DA030396" }, { "agency": "NIH", "grant_number": "DA035942" }, { "agency": "NIH", "grant_number": "DA033831" }, { "agency": "NIH", "grant_number": "DA032464" } ] }, "doi": "10.1371/journal.pone.0188715", "pmcid": "PMC5716591", "primary_object": { "basename": "journal.pone.0188715.pdf", "url": "https://authors.library.caltech.edu/records/7b2js-f0v95/files/journal.pone.0188715.pdf" }, "resource_type": "article", "pub_year": "2017", "author_list": "Parker, Rell L.; O'Neill, Heidi C.; et el." }, { "id": "https://authors.library.caltech.edu/records/wvr6y-jnn54", "eprint_id": 76644, "eprint_status": "archive", "datestamp": "2023-08-19 05:46:19", "lastmod": "2023-10-25 16:10:51", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Henderson-Brandon-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Wall-Teagan-R", "name": { "family": "Wall", "given": "Teagan R." } }, { "id": "Henley-Beverley-M", "name": { "family": "Henley", "given": "Beverley M." } }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "McKinney-Sheri-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Menthol Enhances Nicotine Reward-Related Behavior by Potentiating Nicotine-Induced Changes in nAChR Function, nAChR Upregulation, and DA Neuron Excitability", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2017 The Author(s). This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/. \n\nReceived 20 December 2016. Revised 28 March 2017. Accepted 1 April 2017. Accepted article preview online 12 April 2017. \n\nFunding: National Institutes of Health (NIH) (DA040047, DA033721, DA036061, DA037161, and DA037743). \n\nWe declare no conflict of interest.\n\nPublished - npp201772a.pdf
Supplemental Material - npp201772x1.docx
", "abstract": "Understanding why the quit rate among smokers of menthol cigarettes is lower than non-menthol smokers requires identifying the neurons that are altered by nicotine, menthol, and acetylcholine. Dopaminergic (DA) neurons in the ventral tegmental area (VTA) mediate the positive reinforcing effects of nicotine. Using mouse models, we show that menthol enhances nicotine-induced changes in nicotinic acetylcholine receptors (nAChRs) expressed on midbrain DA neurons. Menthol plus nicotine upregulates nAChR number and function on midbrain DA neurons more than nicotine alone. Menthol also enhances nicotine-induced changes in DA neuron excitability. In a conditioned place preference (CPP) assay, we observed that menthol plus nicotine produces greater reward-related behavior than nicotine alone. Our results connect changes in midbrain DA neurons to menthol-induced enhancements of nicotine reward-related behavior and may help explain how smokers of menthol cigarettes exhibit reduced cessation rates.", "date": "2017-11", "date_type": "published", "publication": "Neuropsychopharmacology", "volume": "42", "number": "12", "publisher": "Nature Publishing Group", "pagerange": "2285-2291", "id_number": "CaltechAUTHORS:20170418-150133796", "issn": "0893-133X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170418-150133796", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA040047" }, { "agency": "NIH", "grant_number": "DA033721" }, { "agency": "NIH", "grant_number": "DA036061" }, { "agency": "NIH", "grant_number": "DA037161" }, { "agency": "NIH", "grant_number": "DA037743" } ] }, "doi": "10.1038/npp.2017.72", "pmcid": "PMC5645749", "primary_object": { "basename": "npp201772a.pdf", "url": "https://authors.library.caltech.edu/records/wvr6y-jnn54/files/npp201772a.pdf" }, "related_objects": [ { "basename": "npp201772x1.docx", "url": "https://authors.library.caltech.edu/records/wvr6y-jnn54/files/npp201772x1.docx" } ], "resource_type": "article", "pub_year": "2017", "author_list": "Henderson, Brandon J.; Wall, Teagan R.; et el." }, { "id": "https://authors.library.caltech.edu/records/z49bw-2zk94", "eprint_id": 83149, "eprint_status": "archive", "datestamp": "2023-08-19 05:44:01", "lastmod": "2023-10-17 22:56:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Tarren-J-R", "name": { "family": "Tarren", "given": "Josephine R." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Belmer-A", "name": { "family": "Belmer", "given": "Arnauld" } }, { "id": "Bartlett-S-E", "name": { "family": "Bartlett", "given": "Selena E." } } ] }, "title": "Acute Ethanol Administration Upregulates Synaptic \u03b14-Subunit of Neuronal Nicotinic Acetylcholine Receptors within the Nucleus Accumbens and Amygdala", "ispublished": "pub", "full_text_status": "public", "keywords": "alcohol, nicotinic receptor, dopamine, nucleus accumbens, amygdala", "note": "\u00a9 2017 Tarren, Lester, Belmer and Bartlett. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. \n\nReceived: 03 July 2017; Accepted: 05 October 2017; Published: 24 October 2017. \n\nAuthor Contributions: HL provided the \u03b14YFP mice and revised the manuscript. JT performed the WB and qPCR experiments and analysis, and drafted the manuscript. AB performed the IHC experiments and analysis. JT, AB, and SB analyzed and interpreted the results. JT, AB, and SB designed the experiments and revised the manuscript. All authors approved the final version of the manuscript. \n\nFunding: National Health and Medical Research Council (NHMRC), GNT104942 and GNT 1061979 to SB; Australian Research Council (ARC) FT1110884 to SB. US National Institutes of Health DA037161 to HL. \n\nThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. \n\nWe thank Ms. Joan Holgate for excellent technical assistance and advice, as well as for animal ethics and breeding. We are grateful to the imaging facility of the Translational Research Institute and the facility manager Sandrine Roy for the extensive use of resources. We also thank Mick Grohuk for his assistance with manuscript formatting. We acknowledge the National Health and Medical Research Council and the Australian Research Council for funding of this work.\n\nPublished - fnmol-10-00338.pdf
Supplemental Material - image_1.tif
Supplemental Material - image_2.tif
", "abstract": "Alcohol and nicotine are two of the most frequently abused drugs, with their comorbidity well described. Previous data show that chronic exposure to nicotine upregulates high-affinity nicotinic acetylcholine receptors (nAChRs) in several brain areas. Effects of ethanol on specific brain nAChR subtypes within the mesolimbic dopaminergic (DA) pathway may be a key element in the comorbidity of ethanol and nicotine. However, it is unknown how alcohol affects the abundance of these receptor proteins. In the present study, we measured the effect of acute binge ethanol on nAChR \u03b14 subunit levels in the prefrontal cortex (PFC), nucleus accumbens (NAc), ventral tegmental area (VTA), and amygdala (Amg) by western blot analysis using a knock-in mouse line, generated with a normally functioning \u03b14 nAChR subunit tagged with yellow fluorescent protein (YFP). We observed a robust increase in \u03b14-YFP subunit levels in the NAc and the Amg following acute ethanol, with no changes in the PFC and VTA. To further investigate whether this upregulation was mediated by increased local mRNA transcription, we quantified mRNA levels of the Chrna4 gene using qRT-PCR. We found no effect of ethanol on \u03b14 mRNA expression, suggesting that the upregulation of \u03b14 protein rather occurs post-translationally. The quantitative counting of YFP immunoreactive puncta further revealed that \u03b14-YFP protein is upregulated in presynaptic boutons of the dopaminergic axons projecting to the shell and the core regions of the NAc as well as to the basolateral amygdala (BLA), but not to the central or lateral Amg. Together, our results demonstrate that a single exposure to binge ethanol upregulates level of synaptic \u03b14\u2217 nAChRs in dopaminergic inputs to the NAc and BLA. This upregulation could be linked to the functional dysregulation of dopaminergic signalling observed during the development of alcohol dependence.", "date": "2017-10-24", "date_type": "published", "publication": "Frontiers in Molecular Neuroscience", "volume": "10", "publisher": "Frontiers Research Foundation", "pagerange": "Art. No. 338", "id_number": "CaltechAUTHORS:20171113-110814227", "issn": "1662-5099", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20171113-110814227", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Health and Medical Research Council (NHMRC)", "grant_number": "GNT 104942" }, { "agency": "National Health and Medical Research Council (NHMRC)", "grant_number": "GNT 1061979" }, { "agency": "Australian Research Council", "grant_number": "FT1110884" }, { "agency": "NIH", "grant_number": "DA037161" } ] }, "doi": "10.3389/fnmol.2017.00338", "pmcid": "PMC5660714", "primary_object": { "basename": "fnmol-10-00338.pdf", "url": "https://authors.library.caltech.edu/records/z49bw-2zk94/files/fnmol-10-00338.pdf" }, "related_objects": [ { "basename": "image_1.tif", "url": "https://authors.library.caltech.edu/records/z49bw-2zk94/files/image_1.tif" }, { "basename": "image_2.tif", "url": "https://authors.library.caltech.edu/records/z49bw-2zk94/files/image_2.tif" } ], "resource_type": "article", "pub_year": "2017", "author_list": "Tarren, Josephine R.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/b0bch-7b106", "eprint_id": 81917, "eprint_status": "archive", "datestamp": "2023-08-19 04:54:44", "lastmod": "2023-10-17 21:55:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Shivange-A-V", "name": { "family": "Shivange", "given": "Amol V." } }, { "id": "Nichols-A-L", "name": { "family": "Nichols", "given": "Aaron L." }, "orcid": "0000-0001-9341-0049" }, { "id": "Borden-P-M", "name": { "family": "Borden", "given": "Philip M." } }, { "id": "Kamjaya-A", "name": { "family": "Kamjaya", "given": "Aron" } }, { "id": "Muthusamy-A-K", "name": { "family": "Muthusamy", "given": "Anand K." }, "orcid": "0000-0003-1041-914X" }, { "id": "Jeon-Janice-H", "name": { "family": "Jeon", "given": "Janice H." }, "orcid": "0000-0002-8483-586X" }, { "id": "Unger-E-K", "name": { "family": "Unger", "given": "Elizabeth K." } }, { "id": "Tian-Lin", "name": { "family": "Tian", "given": "Lin" }, "orcid": "0000-0001-7012-6926" }, { "id": "Marvin-J-S", "name": { "family": "Marvin", "given": "Jonathan S." }, "orcid": "0000-0003-2294-4515" }, { "id": "Looger-L-L", "name": { "family": "Looger", "given": "Loren L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Nicotine in the Endoplasmic Reticulum", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2017 The Rockefeller University Press. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. \n\nPublished September 4, 2017. \n\nSupport: NIH, NARSAD, California TRDRP, and HHMI.\n\nPublished - SHIVANGE_2017p14a.pdf
", "abstract": "Nicotine activates plasma membrane (PM) nicotinic\nreceptors (nAChRs), but also permeates into the endoplasmic\nreticulum (ER) and cis-Golgi, and there binds to nascent nAChRs. Other psychiatric and abused drugs may also enter the ER and bind their classical targets. Further progress requires direct proof, quantification, and time resolution of these processes in live cells and in the brain of animals. Therefore, we are developing genetically encoded fluorescent biosensors to study the subcellular pharmacokinetics of neural drugs.", "date": "2017-09", "date_type": "published", "publication": "Journal of General Physiology", "volume": "149", "number": "9", "publisher": "Rockefeller University Press", "pagerange": "Art. No. 32", "id_number": "CaltechAUTHORS:20170929-092012776", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170929-092012776", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "NARSAD" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Howard Hughes Medical Institute (HHMI)" } ] }, "doi": "10.1085/jgp.149.9.889", "primary_object": { "basename": "SHIVANGE_2017p14a.pdf", "url": "https://authors.library.caltech.edu/records/b0bch-7b106/files/SHIVANGE_2017p14a.pdf" }, "resource_type": "article", "pub_year": "2017", "author_list": "Shivange, Amol V.; Nichols, Aaron L.; et el." }, { "id": "https://authors.library.caltech.edu/records/rxq2x-x2a05", "eprint_id": 82344, "eprint_status": "archive", "datestamp": "2023-08-19 04:55:29", "lastmod": "2023-10-17 22:12:54", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Wall-T-R", "name": { "family": "Wall", "given": "Teagan R." } }, { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Voren-G", "name": { "family": "Voren", "given": "George" } }, { "id": "Wageman-C-R", "name": { "family": "Wageman", "given": "Charles R." } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Yohannes-D", "name": { "family": "Yohannes", "given": "Daniel" } }, { "id": "Kenny-P-J", "name": { "family": "Kenny", "given": "Paul J." } }, { "id": "Bencherif-M", "name": { "family": "Bencherif", "given": "Merouane" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "TC299423, a Novel Agonist for Nicotinic Acetylcholine Receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotine addiction, nicotinic acetylcholine receptors, neuroprotection, electrophysiology, transmitter\nrelease, a6b2^*, hexahydroazocine, pyrimidine", "note": "Copyright \u00a9 2017 Wall, Henderson, Voren, Wageman, Deshpande, Cohen, Grady, Marks, Yohannes, Kenny, Bencherif and Lester. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. \n\nReceived: 25 June 2017. Accepted: 29 August 2017. Published: 26 September 2017. \n\nWe thank Allan C. Collins, William A. Corrigall, John A. Lowe, and Paul Whiteaker for much advice during this research. \n\nAuthor Contributions: Experiments performed by TW, BH, GV, CW, PD, BC, SG, and MM. Analysis by TW, BH, GV, CW, BC, SG, MM, DY, MB, and HL. Research direction by MM, DY, PK, MB, and HL. Manuscript preparation and revision by TW, BH, BC, SG, MM, DY, PK, MB, and HL. Funding obtained by BH, MM, PK, MB, and HL. \n\nThis work was supported by US National Institutes of Health grants DA015663 (MM and SG), DA019375 (HL, MM, and MB), DA033721 (BH), DA020686 (PK), and DA033622 (PK).\n\nConflict of Interest Statement: When the research was conducted, MB and DY were employed by Targacept Inc. Targacept has since merged with Catalyst Biosciences. No entity or person now has any intellectual property, or commercial, or financial interest in TC299423. \n\nThe other authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.\n\nPublished - fphar-08-00641.pdf
Supplemental Material - presentation_1.pdf
", "abstract": "(E)-5-(Pyrimidin-5-yl)-1,2,3,4,7,8-hexahydroazocine (TC299423) is a novel agonist for nicotinic acetylcholine receptors (nAChRs). We examined its efficacy, affinity, and potency for \u03b16\u03b22^\u2217 (\u03b16\u03b22-containing), \u03b14\u03b22^\u2217, and \u03b13\u03b24^\u2217 nAChRs, using [^(125)I]-epibatidine binding, whole-cell patch-clamp recordings, synaptosomal ^(86)Rb^+ efflux, [^3H]-dopamine release, and [^3H]-acetylcholine release. TC299423 displayed an EC_(50) of 30\u201360 nM for \u03b16\u03b22^\u2217 nAChRs in patch-clamp recordings and [^3H]-dopamine release assays. Its potency for \u03b16\u03b22^\u2217 in these assays was 2.5-fold greater than that for \u03b14\u03b22^\u2217, and much greater than that for \u03b13\u03b24^\u2217-mediated [^3H]-acetylcholine release. We observed no major off-target binding on 70 diverse molecular targets. TC299423 was bioavailable after intraperitoneal or oral administration. Locomotor assays, measured with gain-of-function, mutant \u03b16 (\u03b16L9\u2032S) nAChR mice, show that TC299423 elicits \u03b16\u03b22^\u2217 nAChR-mediated responses at low doses. Conditioned place preference assays show that low-dose TC299423 also produces significant reward in \u03b16L9\u2032S mice, and modest reward in WT mice, through a mechanism that probably involves \u03b16(non-\u03b14)\u03b22^\u2217 nAChRs. However, TC299423 did not suppress nicotine self-administration in rats, indicating that it did not block nicotine reinforcement in the dosage range that was tested. In a hot-plate test, TC299423 evoked antinociceptive responses in mice similar to those of nicotine. TC299423 and nicotine similarly inhibited mouse marble burying as a measure of anxiolytic effects. Taken together, our data suggest that TC299423 will be a useful small-molecule agonist for future in vitro and in vivo studies of nAChR function and physiology.", "date": "2017-09", "date_type": "published", "publication": "Frontiers in Pharmacology", "volume": "8", "publisher": "Frontiers Media", "pagerange": "Art. No. 641", "id_number": "CaltechAUTHORS:20171013-114018950", "issn": "1663-9812", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20171013-114018950", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA015663" }, { "agency": "NIH", "grant_number": "DA019375" }, { "agency": "NIH", "grant_number": "DA033721" }, { "agency": "NIH", "grant_number": "DA020686" }, { "agency": "NIH", "grant_number": "DA033622" } ] }, "doi": "10.3389/fphar.2017.00641", "pmcid": "PMC5626944", "primary_object": { "basename": "fphar-08-00641.pdf", "url": "https://authors.library.caltech.edu/records/rxq2x-x2a05/files/fphar-08-00641.pdf" }, "related_objects": [ { "basename": "presentation_1.pdf", "url": "https://authors.library.caltech.edu/records/rxq2x-x2a05/files/presentation_1.pdf" } ], "resource_type": "article", "pub_year": "2017", "author_list": "Wall, Teagan R.; Henderson, Brandon J.; et el." }, { "id": "https://authors.library.caltech.edu/records/kv1wc-4sc65", "eprint_id": 80024, "eprint_status": "archive", "datestamp": "2023-08-21 21:30:54", "lastmod": "2023-10-26 17:31:34", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Mulcahy-Matthew-J", "name": { "family": "Mulcahy", "given": "Matthew J." }, "orcid": "0000-0001-5588-8762" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Granulocytes as models for human protein marker identification following nicotine exposure", "ispublished": "pub", "full_text_status": "public", "keywords": "granulocytes; nAChRs; nicotine", "note": "\u00a9 2017 International Society for Neurochemistry. \n\nIssue online: 8 August 2017; Version of record online: 8 August 2017; Manuscript Accepted: 1 March 2017; Manuscript Revised: 28 February 2017; Manuscript Received: 16 November 2016. \n\nThis work was funded by National Institutes of Health (DA036061). \n\nThe authors have no conflicts of interest to declare.\n\nAccepted Version - nihms982148.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are pentameric cation channels expressed in the mammalian CNS, in the peripheral nervous system, and in skeletal muscle. Neuronal-type nAChRs are also found in several non-neuronal cell types, including leukocytes. Granulocytes are a subtype of leukocytes that include basophils, eosinophils, and neutrophils. Granulocytes, also known as polymorphonuclear leukocytes, are characterized by their ability to produce, store, and release compounds from intracellular granules. Granulocytes are the most abundant type of leukocyte circulating in the peripheral blood. Granulocyte abundance, nAChR expression, and nAChR upregulation following chronic nicotine administration makes granulocytes interesting models for identifying protein markers of nicotine exposure. Nicotinic receptor subunits and several non-nAChR proteins have been identified as protein markers of granulocyte nicotine exposure. We review methods to isolate granulocytes from human tissue, summarize present data about the expression of nAChRs in the three granulocyte cell types (basophils, eosinophils, and neutrophils), describe current knowledge of the effects of nicotine exposure on human granulocyte protein expression, and highlight areas of interest for future investigation.", "date": "2017-08", "date_type": "published", "publication": "Journal of Neurochemistry", "volume": "142", "number": "S2", "publisher": "Blackwell Publishing", "pagerange": "151-161", "id_number": "CaltechAUTHORS:20170809-134755993", "issn": "0022-3042", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170809-134755993", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA036061" } ] }, "doi": "10.1111/jnc.14010", "pmcid": "PMC6057152", "primary_object": { "basename": "nihms982148.pdf", "url": "https://authors.library.caltech.edu/records/kv1wc-4sc65/files/nihms982148.pdf" }, "resource_type": "article", "pub_year": "2017", "author_list": "Mulcahy, Matthew J. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/nan8x-xb833", "eprint_id": 79286, "eprint_status": "archive", "datestamp": "2023-08-19 04:11:15", "lastmod": "2023-10-26 14:46:07", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Hurtado-Zavala-Joaquin-I", "name": { "family": "Hurtado-Zavala", "given": "Joaquin I." } }, { "id": "Ramachandran-Binu", "name": { "family": "Ramachandran", "given": "Binu" } }, { "id": "Ahmed-Saheeb", "name": { "family": "Ahmed", "given": "Saheeb" } }, { "id": "Halder-Rashi", "name": { "family": "Halder", "given": "Rashi" } }, { "id": "Bolleyer-Cristiane", "name": { "family": "Bolleyer", "given": "Christiane" } }, { "id": "Awasthi-Ankit", "name": { "family": "Awasthi", "given": "Ankit" } }, { "id": "Stahlberg-Markus-A", "name": { "family": "Stahlberg", "given": "Markus A." } }, { "id": "Wagener-Robin-J", "name": { "family": "Wagener", "given": "Robin J." } }, { "id": "Anderson-Kristin", "name": { "family": "Anderson", "given": "Kristin" } }, { "id": "Drenan-Ryan-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Miwa-Julie-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Staiger-Jochen-F", "name": { "family": "Staiger", "given": "Jochen F." } }, { "id": "Fischer-Andre", "name": { "family": "Fischer", "given": "Andre" } }, { "id": "Dean-Camin", "name": { "family": "Dean", "given": "Camin" } } ] }, "title": "TRPV1 regulates excitatory innervation of OLM neurons in the hippocampus", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2017 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. \n\nReceived: 14 September 2016; Accepted: 09 May 2017; Published online: 19 July 2017. \n\nWe thank Rory McQuiston, Julie Kauer, Manuela Schmidt, and Nils Brose for insightful discussions and critical review of the manuscript. This work was supported by NIH R44Da032464, R21DA033831, PA Cure 4100068719/Lehigh University Accelerator grants to J.M.M., and by a Sofja Kovalevskaja grant from the Alexander von Humboldt Foundation, European Research Council starting grant SytActivity FP7 260916, Deutsche Forschungsgemeinschaft grants CRC889, DE1951/1 and the Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB), and the Boehringer Ingelheim Foundation, to C.D. \n\nAuthor Contributions: J.I.H.-Z. conceived the research, designed, performed and analysed experiments, and co-wrote the paper. B.R., S.A. and R.H., designed, performed and analysed experiments. C.B., performed experiments and analysed the data. R.J.W. and K.A. performed experiments. M.A.S. designed and optimized experiments. A.A. analysed experiments. R.M.D., H.A.L. and J.M.M. designed experiments, and contributed animal models. J.F.S. and A.F. supervised and funded the work. C.D. conceived the research, designed and analysed experiments, supervised and funded the work and co-wrote the paper. All authors discussed the results and commented on the manuscript. \n\nThe authors declare no competing financial interests.\n\nPublished - ncomms15878.pdf
Supplemental Material - ncomms15878-s1.pdf
Supplemental Material - ncomms15878-s2.pdf
", "abstract": "TRPV1 is an ion channel activated by heat and pungent agents including capsaicin, and has been extensively studied in nociception of sensory neurons. However, the location and function of TRPV1 in the hippocampus is debated. We found that TRPV1 is expressed in oriens-lacunosum-moleculare (OLM) interneurons in the hippocampus, and promotes excitatory innervation. TRPV1 knockout mice have reduced glutamatergic innervation of OLM neurons. When activated by capsaicin, TRPV1 recruits more glutamatergic, but not GABAergic, terminals to OLM neurons in vitro. When TRPV1 is blocked, glutamatergic input to OLM neurons is dramatically reduced. Heterologous expression of TRPV1 also increases excitatory innervation. Moreover, TRPV1 knockouts have reduced Schaffer collateral LTP, which is rescued by activating OLM neurons with nicotine\u2014via \u03b12\u03b22-containing nicotinic receptors\u2014to bypass innervation defects. Our results reveal a synaptogenic function of TRPV1 in a specific interneuron population in the hippocampus, where it is important for gating hippocampal plasticity.", "date": "2017-07-19", "date_type": "published", "publication": "Nature Communications", "volume": "8", "publisher": "Nature Publishing Group", "pagerange": "Art. No. 15878", "id_number": "CaltechAUTHORS:20170724-083914067", "issn": "2041-1723", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170724-083914067", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "R44Da032464" }, { "agency": "NIH", "grant_number": "R21DA033831" }, { "agency": "Lehigh University", "grant_number": "PA Cure 4100068719" }, { "agency": "Alexander von Humboldt Foundation" }, { "agency": "European Research Council (ERC)", "grant_number": "260916" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)", "grant_number": "CRC889" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)", "grant_number": "DE1951/1" }, { "agency": "Center for Nanoscale Microscopy and Molecular Physiology of the Brain (CNMPB)" }, { "agency": "Boehringer Ingelheim Foundation" } ] }, "collection": "CaltechAUTHORS", "doi": "10.1038/ncomms15878", "pmcid": "PMC5524938", "primary_object": { "basename": "ncomms15878.pdf", "url": "https://authors.library.caltech.edu/records/nan8x-xb833/files/ncomms15878.pdf" }, "related_objects": [ { "basename": "ncomms15878-s2.pdf", "url": "https://authors.library.caltech.edu/records/nan8x-xb833/files/ncomms15878-s2.pdf" }, { "basename": "ncomms15878-s1.pdf", "url": "https://authors.library.caltech.edu/records/nan8x-xb833/files/ncomms15878-s1.pdf" } ], "resource_type": "article", "pub_year": "2017", "author_list": "Hurtado-Zavala, Joaquin I.; Ramachandran, Binu; et el." }, { "id": "https://authors.library.caltech.edu/records/cf249-mpp31", "eprint_id": 75083, "eprint_status": "archive", "datestamp": "2023-08-19 02:31:56", "lastmod": "2023-10-25 14:43:37", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Post-Michael-R", "name": { "family": "Post", "given": "Michael R." }, "orcid": "0000-0002-3214-7619" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Probing for and Quantifying Agonist Hydrogen Bonds in \u03b16\u03b22 Nicotinic Acetylcholine Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2017 American Chemical Society. \n\nReceived: March 8, 2017; Published: March 13, 2017. \n\nThis work was supported by the NIH (NS 34407, DA019375). MRP was supported by an NIH/NRSA training grant: 5 T32 GM07616. We thank Merouane Bencherif and Daniel Yohannes (Targacept) for gifts of TC299423.\n\nAccepted Version - acs_2Ebiochem_2E7b00213.pdf
Accepted Version - nihms-982744.pdf
", "abstract": "Designing subtype-selective agonists for neuronal nicotinic acetylcholine receptors (nACh\u00acR) is a challenging and significant goal aided by intricate knowledge of each subtype's binding patterns. We previously reported that in \u03b16\u03b22 receptors, acetylcholine makes a functional cation-\u03c0 interaction with Trp149, but nicotine and TC299423 do not, suggesting a distinctive binding site. This work explores hydrogen binding at the backbone carbonyl associated with \u03b16\u03b22 Trp149. Substituting the i+1 residue, Thr150, with its \u03b1-hydroxy analogue (Tah) attenuates the carbonyl's hydrogen bond accepting ability. At \u03b16(T150Tah)\u03b22, nicotine shows a 24-fold loss of function, TC299423 shows a modest loss, and acetylcholine shows no effect. Nicotine was further analyzed via a double-mutant cycle analysis utilizing N'-methylnicotinium, which indicated a hydrogen bond in \u03b16\u03b22 with a \u0394\u0394G of 2.6 kcal/mol. Thus, even though nicotine does not make the conserved cation-\u03c0 interaction with Trp149, it still makes a functional hydrogen bond to its associated backbone carbonyl.", "date": "2017-04-04", "date_type": "published", "publication": "Biochemistry", "volume": "56", "number": "13", "publisher": "American Chemical Society", "pagerange": "1836-1840", "id_number": "CaltechAUTHORS:20170314-072647704", "issn": "0006-2960", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170314-072647704", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "DA019375" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "T32 GM07616" } ] }, "doi": "10.1021/acs.biochem.7b00213", "pmcid": "PMC6075822", "primary_object": { "basename": "acs_2Ebiochem_2E7b00213.pdf", "url": "https://authors.library.caltech.edu/records/cf249-mpp31/files/acs_2Ebiochem_2E7b00213.pdf" }, "related_objects": [ { "basename": "nihms-982744.pdf", "url": "https://authors.library.caltech.edu/records/cf249-mpp31/files/nihms-982744.pdf" } ], "resource_type": "article", "pub_year": "2017", "author_list": "Post, Michael R.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/0sb0f-0hy75", "eprint_id": 81709, "eprint_status": "archive", "datestamp": "2023-08-19 02:28:21", "lastmod": "2023-10-17 21:11:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Mulcahy-M-J", "name": { "family": "Mulcahy", "given": "Matthew J." }, "orcid": "0000-0001-5588-8762" }, { "id": "Wang-Jonathan-H", "name": { "family": "Wang", "given": "Jonathan H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Region specific proteomic analysis of murine brain after chronic nicotine or menthol", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2017 by the Federation of American Societies for Experimental Biology. \n\nThis work is supported by NIH DA036061.", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are pentameric cation channels that are expressed in the mammalian central nervous system, the peripheral nervous system, and the neuromuscular junction. Eleven neuronal nAChR subunits have been identified in mammals (\u03b12-7, \u03b19-10, \u03b22-4). Chronic administration of nicotine has been shown to increase total \u03b14 and \u03b22 subunit expression in specific brain regions. Menthol, a common cigarette flavoring, has also been shown to upregulate \u03b14 and \u03b22 subunit expression in the cortex, hippocampus, and striatum independent of nicotine co-administration. These data highlight the potential use of changes in \u03b22 nAChR subunit expression as a marker for neuronal nicotine exposure when administered alone and when administered with additional bioactive compounds such as menthol. We investigated the effect of nicotine and of menthol in several murine brain regions on changes in total \u03b22 subunit protein expression using western blot analysis. Absence of \u03b22 immunoreactivity was confirmed in both \u03b22 knockout and \u03b14 knockout mice. To assess the effects of nicotine, male C57bl/6 mice were separated into two treatment groups: control (saline) and nicotine (2mg/kg/hr). Vehicle (ethanol) and menthol alone (2mg/kg/hr) were compared separately. All treatments were administered for 10\u201312 days. Nicotine-induced \u03b22 immunoreactivity was significantly upregulated in isolated brain regions, including the cortex, hippocampus, midbrain, striatum, and thalamus. No change in immunoreactivity was observed in the habenula of mice treated chronically with nicotine. In response to chronic menthol treatment, a significant increase was only observed in the hypothalamus. These data indicate that nicotine and menthol differentially effect expression of nAChR subtypes in various regions of the brain. A comprehensive analysis of changes in \u03b22 nAChR subunit expression in various brain regions will facilitate our understanding of the effects of nicotine exposure in the mammalian CNS. Additionally, the validation of a reliable marker for nicotine and menthol exposure in regions associated with various aspects of addiction and dependence would be invaluable in the pursuit of better cessation therapeutics. Using \u03b22 expression as a marker of nicotine exposure will enable a better understanding of possible effects of nicotine and menthol.", "date": "2017-04", "date_type": "published", "publication": "FASEB Journal", "volume": "31", "number": "S1", "publisher": "Federation of American Societies for Experimental Biology", "pagerange": "Art. No. 991.1", "id_number": "CaltechAUTHORS:20170921-153023998", "issn": "0892-6638", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170921-153023998", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA036061" } ] }, "resource_type": "article", "pub_year": "2017", "author_list": "Mulcahy, Matthew J.; Wang, Jonathan H.; et el." }, { "id": "https://authors.library.caltech.edu/records/x6w4k-e8875", "eprint_id": 78095, "eprint_status": "archive", "datestamp": "2023-08-22 19:48:02", "lastmod": "2023-10-25 23:45:13", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Post-Michael-R", "name": { "family": "Post", "given": "Michael R." }, "orcid": "0000-0002-3214-7619" }, { "id": "Tender-Gabrielle-S", "name": { "family": "Tender", "given": "Gabrielle S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Secondary Ammonium Agonists Make Dual Cation-\u03c0 Interactions in \u03b14\u03b22 Nicotinic Receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "Parkinson's disease; Addiction; Ion channels; Nicotinic acetylcholine receptors; Electrophysiology; Non-canonical amino acids", "note": "\u00a9 2017 by the Society for Neuroscience. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. \n\nReceived January 26, 2017; accepted March 2, 2017; First published March 17, 2017. \n\nThis work was supported by HHS | NIH | National Institute of Neurological Disorders and Stroke (NINDS) (100000065, Grant NS34407); HHS | NIH | National Institute on Drug Abuse (NIDA) (Funding 100000026, Grand DA 019375); and by Beckman Institute at Caltech. M.R.P. was supported by the NIH/NRSA Training Grant 5 T32 GM07616. \n\nThe authors declare no competing financial interests.\n\nPublished - ENEURO.0032-17.2017.pdf
", "abstract": "A cation-\u03c0 interaction between the ammonium group of an agonist and a conserved tryptophan termed TrpB is a near universal feature of agonist binding to nicotinic acetylcholine receptors (nAChRs). TrpB is one of five residues that form the aromatic box of the agonist binding site, and for the prototype agonists ACh and nicotine, only TrpB makes a functional cation-\u03c0 interaction. We report that, in addition to TrpB, a significant cation-\u03c0 interaction is made to a second aromatic, TyrC2, by the agonists metanicotine, TC299423, varenicline, and nornicotine. A common structural feature of these agonists, and a distinction from ACh and nicotine, is a protonated secondary amine that provides the cation for the cation-\u03c0 interaction. These results indicate a distinction in binding modes between agonists with subtly different structures that may provide guidance for the development of subtype-selective agonists of nAChRs.", "date": "2017-03", "date_type": "published", "publication": "eNeuro", "volume": "4", "number": "2", "publisher": "Society for Neuroscience", "pagerange": "Art. No. 0032-17", "id_number": "CaltechAUTHORS:20170612-100040551", "issn": "2373-2822", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170612-100040551", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "DA 019375" }, { "agency": "Caltech Beckman Institute" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5 T32 GM07616" } ] }, "doi": "10.1523/ENEURO.0032-17.2017", "pmcid": "PMC5458768", "primary_object": { "basename": "ENEURO.0032-17.2017.pdf", "url": "https://authors.library.caltech.edu/records/x6w4k-e8875/files/ENEURO.0032-17.2017.pdf" }, "resource_type": "article", "pub_year": "2017", "author_list": "Post, Michael R.; Tender, Gabrielle S.; et el." }, { "id": "https://authors.library.caltech.edu/records/jvj0f-q0643", "eprint_id": 75239, "eprint_status": "archive", "datestamp": "2023-08-19 01:29:42", "lastmod": "2023-10-25 14:52:45", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Henley-B-M", "name": { "family": "Henley", "given": "Beverley M." } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "Gold-H-D", "name": { "family": "Gold", "given": "Heather D." } }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Reliable Identification of Living Dopaminergic Neurons in Midbrain Cultures Using RNA Sequencing and TH-promoter-driven eGFP Expression", "ispublished": "pub", "full_text_status": "public", "keywords": "Neuroscience, Issue 120, single-cell, RNA-sequencing, cell harvesting, midbrain cultures, dopaminergic neurons, Parkinson's disease", "note": "\u00a9 JoVE 2017. \n\n\nDate Published: 2/10/2017.\n\nThis work was supported by grants from the U.S. National Institutes of Health (DA017279, AG033954, DA037743), the Michael J. Fox Foundation, and the Caltech Innovation Initiative funding the Millard and Muriel Jacobs Genetics and Genomics Laboratory at the California Institute of Technology. We thank Brian Williams for optimising the RNA-Seq library protocol and for providing assistance. We thank Henry Amrhein for computational training. We thank Barbara J. Wold for the use of her equipment and laboratory space. We thank Igor Antoshechkin for library sequencing, and for facility management. \n\nThe authors have nothing to disclose.\n\nPublished - jove-protocol-54981-reliable-identification-living-dopaminergic-neurons-midbrain-cultures.pdf
", "abstract": "In Parkinson's Disease (PD) there is widespread neuronal loss throughout the brain with pronounced degeneration of dopaminergic neurons in the SNc, leading to bradykinesia, rigidity, and tremor. The identification of living dopaminergic neurons in primary Ventral Mesencephalic (VM) cultures using a fluorescent marker provides an alternative way to study the selective vulnerability of these neurons without relying on the immunostaining of fixed cells. Here, we isolate, dissociate, and culture mouse VM neurons for 3 weeks. We then identify dopaminergic neurons in the cultures using eGFP fluorescence (driven by a Tyrosine Hydroxylase (TH) promoter). Individual neurons are harvested into microcentrifuge tubes using glass micropipettes. Next, we lyse the harvested cells, and conduct cDNA synthesis and transposon-mediated \"tagmentation\" to produce single cell RNA-Seq libraries. After passing a quality-control check, single-cell libraries are sequenced and subsequent analysis is carried out to measure gene expression. We report transcriptome results for individual dopaminergic and GABAergic neurons isolated from midbrain cultures. We report that 100% of the live TH-eGFP cells that were harvested and sequenced were dopaminergic neurons. These techniques will have widespread applications in neuroscience and molecular biology.", "date": "2017-02-10", "date_type": "published", "publication": "Journal of Visualized Experiments", "number": "120", "publisher": "JoVE", "pagerange": "Art. No. 54981", "id_number": "CaltechAUTHORS:20170320-104204700", "issn": "1940-087X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170320-104204700", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "DA037743" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "Caltech Innovation Initiative (CI2)" } ] }, "doi": "10.3791/54981", "pmcid": "PMC5409190", "primary_object": { "basename": "jove-protocol-54981-reliable-identification-living-dopaminergic-neurons-midbrain-cultures.pdf", "url": "https://authors.library.caltech.edu/records/jvj0f-q0643/files/jove-protocol-54981-reliable-identification-living-dopaminergic-neurons-midbrain-cultures.pdf" }, "resource_type": "article", "pub_year": "2017", "author_list": "Henley, Beverley M.; Cohen, Bruce N.; et el." }, { "id": "https://authors.library.caltech.edu/records/je69j-s5c48", "eprint_id": 69359, "eprint_status": "archive", "datestamp": "2023-08-20 13:15:16", "lastmod": "2023-10-20 16:49:59", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kim-Jinho", "name": { "family": "Kim", "given": "Jinho" }, "orcid": "0000-0001-8711-960X" }, { "id": "Henley-Beverley-M", "name": { "family": "Henley", "given": "Beverley M." }, "orcid": "0000-0002-6211-2433" }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Yang-Changhuei", "name": { "family": "Yang", "given": "Changhuei" }, "orcid": "0000-0001-8791-0354" } ] }, "title": "Incubator embedded cell culture imaging system (EmSight) based on Fourier ptychographic microscopy", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2016 Optical Society of America. \n\nReceived 23 Apr 2016; revised 7 Jul 2016; accepted 17 Jul 2016; published 22 Jul 2016. \n\nFunding: National Institute of Health (NIH) (R01AI6226-01); Caltech Agency Award (AMGEN.96EYES). \n\nWe would like to thank Xiaoze Ou and Jaebum Chung for helpful discussions, and Daniel Martin for fabrication of a Siemens star phase target. Jinho Kim's work was supported by the Kwanjeong Educational Foundation.\n\nPublished - boe-7-8-3097.pdf
", "abstract": "Multi-day tracking of cells in culture systems can provide valuable information in bioscience experiments. We report the development of a cell culture imaging system, named EmSight, which incorporates multiple compact Fourier ptychographic microscopes with a standard multiwell imaging plate. The system is housed in an incubator and presently incorporates six microscopes. By using the same low magnification objective lenses as the objective and the tube lens, the EmSight is configured as a 1:1 imaging system that, providing large field-of-view (FOV) imaging onto a low-cost CMOS imaging sensor. The EmSight improves the image resolution by capturing a series of images of the sample at varying illumination angles; the instrument reconstructs a higher-resolution image by using the iterative Fourier ptychographic algorithm. In addition to providing high-resolution brightfield and phase imaging, the EmSight is also capable of fluorescence imaging at the native resolution of the objectives. We characterized the system using a phase Siemens star target, and show four-fold improved coherent resolution (synthetic NA of 0.42) and a depth of field of 0.2 mm. To conduct live, long-term dopaminergic neuron imaging, we cultured ventral midbrain from mice driving eGFP from the tyrosine hydroxylase promoter. The EmSight system tracks movements of dopaminergic neurons over a 21 day period.", "date": "2016-08-01", "date_type": "published", "publication": "Biomedical Optics Express", "volume": "7", "number": "8", "publisher": "Optical Society of America", "pagerange": "3097-3110", "id_number": "CaltechAUTHORS:20160801-135931634", "issn": "2156-7085", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160801-135931634", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "R01AI6226-01" }, { "agency": "Caltech", "grant_number": "AMGEN.96EYES" }, { "agency": "Kwanjeong Educational Foundation" } ] }, "doi": "10.1364/BOE.7.003097", "pmcid": "PMC4986817", "primary_object": { "basename": "boe-7-8-3097.pdf", "url": "https://authors.library.caltech.edu/records/je69j-s5c48/files/boe-7-8-3097.pdf" }, "resource_type": "article", "pub_year": "2016", "author_list": "Kim, Jinho; Henley, Beverley M.; et el." }, { "id": "https://authors.library.caltech.edu/records/f5g8q-s5157", "eprint_id": 68854, "eprint_status": "archive", "datestamp": "2023-08-20 11:51:10", "lastmod": "2023-10-20 15:48:54", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nichols-W-A", "name": { "family": "Nichols", "given": "Weston A." } }, { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Marotta-C-B", "name": { "family": "Marotta", "given": "Christopher B." } }, { "id": "Yu-Caroline-Y", "name": { "family": "Yu", "given": "Caroline Y." } }, { "id": "Richards-C", "name": { "family": "Richards", "given": "Chris" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } } ] }, "title": "Mutation Linked to Autosomal Dominant Nocturnal Frontal Lobe Epilepsy Reduces Low-Sensitivity \u03b14\u03b22, and Increases \u03b15\u03b14\u03b22, Nicotinic Receptor Surface Expression", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2016 Nichols et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. \n\nReceived: February 22, 2016; Accepted: June 9, 2016; Published: June 23, 2016. \n\nThis research was supported by grants to D.A.D. (NS034407), H.A.L. (DA017279), B.J.H. (DA033721), and C.R. (DA030877) from the National Institutes of Health (http://www.nih.gov/) and to C.Y.Y. from the Rose Hills foundation (http://rosehillsfoundation.org/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. \n\nThe authors acknowledge the assistance of Timothy F. Miles (Division of Biology & Biological Engineering, California Institute of Technology, Pasadena, CA) in making the TIRF measurements. \n\nAuthor Contributions: Conceived and designed the experiments: WAN BJH CBM HAL BNC. Performed the experiments: WAN BJH CBM CYY. Analyzed the data: WAN BJH CBM CYY BNC. Contributed reagents/materials/analysis tools: CR. Wrote the paper: WAN BJH CBM DAD HAL BNC. \n\nData Availability: All relevant data are within the paper. \n\nThe authors have declared that no competing interests exist.\n\nPublished - PloSe0158032.pdf
", "abstract": "A number of mutations in \u03b14\u03b22-containing (\u03b14\u03b22*) nicotinic acetylcholine (ACh) receptors (nAChRs) are linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE), including one in the \u03b22 subunit called \u03b22V287L. Two \u03b14\u03b22* subtypes with different subunit stoichiometries and ACh sensitivities co-exist in the brain, a high-sensitivity subtype with (\u03b14)2(\u03b22)3 subunit stoichiometry and a low-sensitivity subtype with (\u03b14)3(\u03b22)2 stoichiometry. The \u03b15 nicotinic subunit also co-assembles with \u03b14\u03b22 to form a high-sensitivity \u03b15\u03b14\u03b22 nAChR. Previous studies suggest that the \u03b22V287L mutation suppresses low-sensitivity \u03b14\u03b22* nAChR expression in a knock-in mouse model and also that \u03b15 co-expression improves the surface expression of ADNFLE mutant nAChRs in a cell line. To test these hypotheses further, we expressed mutant and wild-type (WT) nAChRs in oocytes and mammalian cell lines, and measured the effects of the \u03b22V287L mutation on surface receptor expression and the ACh response using electrophysiology, a voltage-sensitive fluorescent dye, and superecliptic pHluorin (SEP). The \u03b22V287L mutation reduced the EC_(50) values of high- and low-sensitivity \u03b14\u03b22 nAChRs expressed in Xenopus oocytes for ACh by a similar factor and suppressed low-sensitivity \u03b14\u03b22 expression. In contrast, it did not affect the EC50 of \u03b15\u03b14\u03b22 nAChRs for ACh. Measurements of the ACh responses of WT and mutant nAChRs expressed in mammalian cell lines using a voltage-sensitive fluorescent dye and whole-cell patch-clamping confirm the oocyte data. They also show that, despite reducing the maximum response, \u03b22V287L increased the \u03b14\u03b22 response to a sub-saturating ACh concentration (1 \u03bcM). Finally, imaging SEP-tagged \u03b15, \u03b14, \u03b22, and \u03b22V287L subunits showed that \u03b22V287L reduced total \u03b14\u03b22 nAChR surface expression, increased the number of \u03b22 subunits per \u03b14\u03b22 receptor, and increased surface \u03b15\u03b14\u03b22 nAChR expression. Thus, the \u03b22V287L mutation alters the subunit composition and sensitivity of \u03b14\u03b22 nAChRs, and increases \u03b15\u03b14\u03b22 surface expression.", "date": "2016-06", "date_type": "published", "publication": "PLoS ONE", "volume": "11", "number": "6", "publisher": "Public Library of Science", "pagerange": "Art. No. e0158032", "id_number": "CaltechAUTHORS:20160706-102113506", "issn": "1932-6203", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160706-102113506", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "DA033721" }, { "agency": "NIH", "grant_number": "DA030877" }, { "agency": "Rose Hills Foundation" } ] }, "doi": "10.1371/journal.pone.0158032", "pmcid": "PMC4918917", "primary_object": { "basename": "PloSe0158032.pdf", "url": "https://authors.library.caltech.edu/records/f5g8q-s5157/files/PloSe0158032.pdf" }, "resource_type": "article", "pub_year": "2016", "author_list": "Nichols, Weston A.; Henderson, Brandon J.; et el." }, { "id": "https://authors.library.caltech.edu/records/dm1gj-z2c98", "eprint_id": 65359, "eprint_status": "archive", "datestamp": "2023-08-22 17:34:03", "lastmod": "2023-10-18 14:34:35", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Henderson-Brandon-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Wall-Teagan-R", "name": { "family": "Wall", "given": "Teagan R." } }, { "id": "Henley-Beverley-M", "name": { "family": "Henley", "given": "Beverley M." }, "orcid": "0000-0002-6211-2433" }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "Nichols-Weston-A", "name": { "family": "Nichols", "given": "Weston A." } }, { "id": "Moaddel-Ruin", "name": { "family": "Moaddel", "given": "Ruin" }, "orcid": "0000-0002-6812-0127" }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Menthol Alone Upregulates Midbrain nAChRs, Alters nAChR Subtype Stoichiometry, Alters Dopamine Neuron Firing Frequency, and Prevents Nicotine Reward", "ispublished": "pub", "full_text_status": "public", "keywords": "dopamine neuron; electrophysiology; menthol; nicotine; nicotinic receptor; reward", "note": "\u00a9 2016 the authors. For the first six months after publication SfN's license will be exclusive. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). \n\nReceived Nov. 21, 2015; revised Jan. 19, 2016; accepted Jan. 29, 2016. \n\nThis work was supported by the National Institutes of Health (NIH; DA017279, DA019375, DA033721, DA036061, DA037161, and DA037743), the National Institute on Aging (NIH) Intramural Research Program, and the California Tobacco-Related Disease Research Program (17RT0127). We thank Michael Marks, Sharon Grady, and Ying Xie for menthol assays. We thank Heather Gold for help with bioinformatics. \n\nAuthor contributions: B.J.H., C.X., and H.A.L. designed research; B.J.H., T.R.W., B.M.H., C.H.K., and W.A.N. performed research; B.J.H., C.H.K., and R.M. contributed unpublished reagents/analytic tools; B.J.H., T.R.W., B.M.H., and W.A.N. analyzed data; B.J.H., B.M.H., C.X., and H.A.L. wrote the paper. \n\nThe authors declare no competing financial interests.\n\nPublished - 2957.full.pdf
", "abstract": "Upregulation of \u03b22 subunit-containing (\u03b22*) nicotinic acetylcholine receptors (nAChRs) is implicated in several aspects of nicotine addiction, and menthol cigarette smokers tend to upregulate \u03b22* nAChRs more than nonmenthol cigarette smokers. We investigated the effect of long-term menthol alone on midbrain neurons containing nAChRs. In midbrain dopaminergic (DA) neurons from mice containing fluorescent nAChR subunits, menthol alone increased the number of \u03b14 and \u03b16 nAChR subunits, but this upregulation did not occur in midbrain GABAergic neurons. Thus, chronic menthol produces a cell-type-selective upregulation of \u03b14* nAChRs, complementing that of chronic nicotine alone, which upregulates \u03b14 subunit-containing (\u03b14*) nAChRs in GABAergic but not DA neurons. In mouse brain slices and cultured midbrain neurons, menthol reduced DA neuron firing frequency and altered DA neuron excitability following nAChR activation. Furthermore, menthol exposure before nicotine abolished nicotine reward-related behavior in mice. In neuroblastoma cells transfected with fluorescent nAChR subunits, exposure to 500 nM menthol alone also increased nAChR number and favored the formation of (\u03b14)_3(\u03b22)_2 nAChRs; this contrasts with the action of nicotine itself, which favors (\u03b14)_2(\u03b22)_3 nAChRs. Menthol alone also increases the number of \u03b16\u03b22 receptors that exclude the \u03b23 subunit. Thus, menthol stabilizes lower-sensitivity \u03b14* and \u03b16 subunit-containing nAChRs, possibly by acting as a chemical chaperone. The abolition of nicotine reward-related behavior may be mediated through menthol's ability to stabilize lower-sensitivity nAChRs and alter DA neuron excitability. We conclude that menthol is more than a tobacco flavorant: administered alone chronically, it alters midbrain DA neurons of the nicotine reward-related pathway.", "date": "2016-03-09", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "36", "number": "10", "publisher": "Society for Neuroscience", "pagerange": "2957-2974", "id_number": "CaltechAUTHORS:20160315-105822806", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160315-105822806", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "DA019375" }, { "agency": "NIH", "grant_number": "DA033721" }, { "agency": "NIH", "grant_number": "DA036061" }, { "agency": "NIH", "grant_number": "DA037161" }, { "agency": "NIH", "grant_number": "DA037743" }, { "agency": "National Institute on Aging" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "17RT0127" } ] }, "doi": "10.1523/JNEUROSCI.4194-15.2016", "pmcid": "PMC4783498", "primary_object": { "basename": "2957.full.pdf", "url": "https://authors.library.caltech.edu/records/dm1gj-z2c98/files/2957.full.pdf" }, "resource_type": "article", "pub_year": "2016", "author_list": "Henderson, Brandon J.; Wall, Teagan R.; et el." }, { "id": "https://authors.library.caltech.edu/records/9mkk1-zrq61", "eprint_id": 67695, "eprint_status": "archive", "datestamp": "2023-08-20 10:23:58", "lastmod": "2023-10-18 21:24:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Patowary-S", "name": { "family": "Patowary", "given": "Suparna" } }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri L." } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Biener-G", "name": { "family": "Biener", "given": "Gabriel" } }, { "id": "Raicu-V", "name": { "family": "Raicu", "given": "Valerica" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Effects of Menthol on \u03b13\u03b24\u2217 Nicotinic Receptors", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2016 Biophysical Society. Published by Elsevier Inc.", "abstract": "Manufacturers add menthol to roughly 30% of tobacco cigarettes sold in the US, and to an unknown fraction of other tobacco products and electronic cigarettes. Smokers of menthol cigarettes find it harder to quit smoking, raising questions about the mechanism of this apparently harmful menthol effect. Menthol modestly affects the pharmacokinetics of nicotine and acts as a chemical chaperone for nicotinic acetylcholine receptors (nAChRs) [1]. In this study, we have investigated the effects of menthol on \u03b13\u03b24\u2217 nAChRs, which are mostly expressed in medial habenula (MHb) - interpeduncular nucleus (IPN) circuit: a key mediator of nicotine's aversive properties and withdrawal [2]. Fluorescently labeled \u03b13 and \u03b24 subunits are transiently expressed in Neuro-2a cells to form monomers, and oligomers. Using a F\u00f6rster Resonance Energy Transfer (FRET) micro-spectroscopy method [3], we have found that menthol up-regulates \u03b13 subunit while having no effects on \u03b24 subunit and \u03b13\u03b24 pentamer numbers. Our results also show that menthol favors (\u03b13)3(\u03b24)2 stoichiometry over (\u03b13)2(\u03b24)3 stoichiometry. However, the study using Total Internal Reflection Fluorescence Microscopy (TIRFM) reveals that though menthol up-regulates \u03b13 subunit numbers in peripheral Endoplasmic Reticulum, it decreases the \u03b13\u03b24 receptor numbers at the plasma membrane.", "date": "2016-02-16", "date_type": "published", "publication": "Biophysical Journal", "volume": "110", "number": "3", "publisher": "Biophysical Society", "pagerange": "603A", "id_number": "CaltechAUTHORS:20160606-140410523", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160606-140410523", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/j.bpj.2015.11.3221", "resource_type": "article", "pub_year": "2016", "author_list": "Patowary, Suparna; Mackey, Elisha D. W.; et el." }, { "id": "https://authors.library.caltech.edu/records/2rz9x-4d586", "eprint_id": 67694, "eprint_status": "archive", "datestamp": "2023-08-20 10:23:52", "lastmod": "2023-10-18 21:24:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Post-M-R", "name": { "family": "Post", "given": "Michael R." }, "orcid": "0000-0002-3214-7619" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Probing Binding Interactions of Agonists with the \u03b16\u03b22 Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2016 Biophysical Society. Published by Elsevier Inc.", "abstract": "The nicotinic acetylcholine receptor (nAChR) is a ligand gated ion channel and a member of the Cys-loop family, which also contains receptors for serotonin, glycine, and GABA. There are twelve known neuronal subunits which pentamerize in different combinations to form various subtypes, each with a unique function, pharmacology and distribution in the brain. The \u03b16\u03b22-containing subtypes are found mainly in dopaminergic neurons and are thus important targets in the study of Parkinson's disease and addiction. Advanced knowledge of the binding site, which sits at the interface of \u03b16 and \u03b22, could lead toward design of agonists that specifically target this subtype. The \u03b16L9's\u03b22_(LFM/AAQA)L9's (\u03b16\u03b22\u2021) construct enabled heterologous expression and activation of a pure and stoichiometrically controlled population of \u03b16\u03b22 receptors in X. laevis oocytes. Currents were high enough in the \u03b16\u03b22\u2021 system to tolerate nonsense suppression-based non-canonical amino acid mutagenesis, which allowed for structure function studies between the receptor and several agonists.\nInitial structure-function studies probed for a cation-\u03c0 interaction between agonists and the indole side chain of \u03b16 TrpB. This is accomplished using an analog-tryptophan series whereby the negative electrostatic potential on the surface the indole ring was incrementally decreased via electron-withdrawing substituents such as fluorine and measuring the effect that has on binding. Results show that ACh makes a cation-\u03c0 interaction, but surprisingly nicotine and TC-299423 do not. However, the latter two agonists do make a hydrogen bond interaction with the carbonyl of TrpB. This conclusion was made by substituting the adjacent amino acid with an alpha-hydroxy acid and measuring the corresponding shift in binding due to that amide-to-ester backbone mutation. The result was further quantified for nicotine in a double-mutant cycle analysis utilizing N'-methylnicotinium, which showed the interaction to have a coupling energy of \u223c2 kcal/mol.", "date": "2016-02-16", "date_type": "published", "publication": "Biophysical Journal", "volume": "110", "number": "3", "publisher": "Biophysical Society", "pagerange": "603A", "id_number": "CaltechAUTHORS:20160606-140109728", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160606-140109728", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/j.bpj.2015.11.3220", "resource_type": "article", "pub_year": "2016", "author_list": "Post, Michael R.; Dougherty, Dennis A.; et el." }, { "id": "https://authors.library.caltech.edu/records/dw0rx-9k845", "eprint_id": 63525, "eprint_status": "archive", "datestamp": "2023-08-20 09:52:04", "lastmod": "2023-10-25 23:53:22", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Srinivasan-Rahul", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Henley-Beverley-M", "name": { "family": "Henley", "given": "Beverley M." }, "orcid": "0000-0002-6211-2433" }, { "id": "Henderson-Brandon-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Indersmitten-Tim", "name": { "family": "Indersmitten", "given": "Tim" } }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce N." }, "orcid": "0000-0003-2913-6238" }, { "id": "Kim-Charlene-H", "name": { "family": "Kim", "given": "Charlene H." } }, { "id": "McKinney-Sheri-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Deshpande-Purnima", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Smoking-Relevant Nicotine Concentration Attenuates the Unfolded Protein Response in Dopaminergic Neurons", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2016 the authors. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). \n\nReceived June 2, 2015; revised Nov. 9, 2015; accepted Nov. 13, 2015. \n\nThis work was supported by grants from the U.S. National Institutes of Health (AG033954), the Michael J. Fox Foundation, and Louis and Janet Fletcher. We thank Dr. Marie-Fran\u00e7oise Chesselet for useful discussions and E. Mackey for help with antibody verification. \n\nThe authors declare no competing financial interests. \n\nAuthor contributions: R.S., B.M.H., B.J.H., C.X., and H.A.L. designed research; R.S., B.M.H., B.J.H., T.I., B.N.C., C.H.K., S.M., P.D., and C.X. performed research; R.S., B.M.H., B.J.H. contributed unpublished reagents/analytic tools; R.S., B.M.H., B.J.H., T.I., B.N.C., C.X., and H.A.L. analyzed data; R.S., B.M.H., B.J.H., B.N.C., and H.A.L. wrote the paper. \n\nR.S. and B.M.H. contributed equally to this work.\n\nPublished - 65.full.pdf
", "abstract": "Retrospective epidemiological studies show an inverse correlation between susceptibility to Parkinson's disease and a person's history of tobacco use. Animal model studies suggest nicotine as a neuroprotective agent and nicotinic acetylcholine (ACh) receptors (nAChRs) as targets for neuroprotection, but the underlying neuroprotective mechanism(s) are unknown. We cultured mouse ventral midbrain neurons for 3 weeks. Ten to 20% of neurons were dopaminergic (DA), revealed by tyrosine hydroxylase (TH) immunoreactivity. We evoked mild endoplasmic reticulum (ER) stress with tunicamycin (Tu), producing modest increases in the level of nuclear ATF6, phosphorylated eukaryotic initiation factor 2\u03b1, nuclear XBP1, and the downstream proapoptotic effector nuclear C/EBP homologous protein. We incubated cultures for 2 weeks with 200 nm nicotine, the approximate steady-state concentration between cigarette smoking or vaping, or during nicotine patch use. Nicotine incubation suppressed Tu-induced ER stress and the unfolded protein response (UPR). Study of mice with fluorescent nAChR subunits showed that the cultured TH+ neurons displayed \u03b14, \u03b16, and \u03b23 nAChR subunit expression and ACh-evoked currents. Gene expression profile in cultures from TH-eGFP mice showed that the TH+ neurons also express several other genes associated with DA release. Nicotine also upregulated ACh-induced currents in DA neurons by \u223c2.5-fold. Thus, nicotine, at a concentration too low to activate an appreciable fraction of plasma membrane nAChRs, induces two sequelae of pharmacological chaperoning in the ER: UPR suppression and nAChR upregulation. Therefore, one mechanism of neuroprotection by nicotine is pharmacological chaperoning, leading to UPR suppression. Measuring this pathway may help in assessing neuroprotection.", "date": "2016-01-06", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "36", "number": "1", "publisher": "Society for Neuroscience", "pagerange": "65-79", "id_number": "CaltechAUTHORS:20160111-082423100", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160111-082423100", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "Louis and Janet Fletcher" } ] }, "doi": "10.1523/JNEUROSCI.2126-15.2016", "pmcid": "PMC4701966", "primary_object": { "basename": "65.full.pdf", "url": "https://authors.library.caltech.edu/records/dw0rx-9k845/files/65.full.pdf" }, "resource_type": "article", "pub_year": "2016", "author_list": "Srinivasan, Rahul; Henley, Beverley M.; et el." }, { "id": "https://authors.library.caltech.edu/records/vt0s6-vwq82", "eprint_id": 59022, "eprint_status": "archive", "datestamp": "2023-08-20 08:58:26", "lastmod": "2023-10-23 19:58:39", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Lavis-L-D", "name": { "family": "Lavis", "given": "Luke D." }, "orcid": "0000-0002-0789-6343" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Ketamine Inside Neurons?", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2015 American Psychiatric Association. \n\nAccepted: March 1, 2015. \n\nThe authors report no financial relationships with commercial interests.\n\nAccepted Version - nihms-982733.pdf
", "abstract": "Clinically used antidepressants, such as selective serotonin reuptake inhibitors (SSRIs), aid only a fraction of patients. Furthermore, even successful use of SSRIs takes 2 to 6 weeks of maintained medication. Depressed patients need faster help. Since 2000, several clinical studies report that depressed patients given subanesthetic doses of ketamine showed improvement within 2 hours. Trials continue for various dosing regimens, formulations, and populations. It is not understood what causes the therapeutic action of the SSRIs, and it is also not clear how ketamine exerts its effects.\n\nThe best-known behavioral effect of ketamine is dissociative anesthesia. The drug retains Food and Drug Administration (FDA) approval for anesthesia in special populations as well as for veterinary use. The dissociative effects presumably arise from ketamine's action to block N-methyl-D-aspartate (NMDA) receptor channels that have been opened by glutamate. The kinetics, equilibrium, and voltage sensitivity of open-channel blockers is a well-studied topic, and recent work shows how ketamine becomes trapped within the channel pore of NMDA receptors at local concentrations of \u223c1 \u00b5M, which are expected to occur at the clinically effective antidepressant human doses.\n\nHow might blockade of NMDA receptors lead to the antidepressant effects? Most studies emphasize signal transduction pathways that could be modulated by the locally decreased Ca^(2+) flux through NMDA receptors, especially extrasynaptic GluN2B subunit-containing NMDA receptors. In one series of experiments, the decreased Ca^(2+) flux led to decreased activity of eukaryotic elongation factor 2 kinase, which in turn desuppressed eukaryotic elongation factor 2. This ribosome-binding protein then increased translation of brain-derived neurotrophic factor (BDNF). Many other experiments show that BDNF is released during antidepressant action.", "date": "2015-11-01", "date_type": "published", "publication": "American Journal of Psychiatry", "volume": "172", "number": "11", "publisher": "American Psychiatric Publishing", "pagerange": "1064-1066", "id_number": "CaltechAUTHORS:20150728-082757273", "issn": "0002-953X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150728-082757273", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1176/appi.ajp.2015.14121537", "pmcid": "PMC6107348", "primary_object": { "basename": "nihms-982733.pdf", "url": "https://authors.library.caltech.edu/records/vt0s6-vwq82/files/nihms-982733.pdf" }, "resource_type": "article", "pub_year": "2015", "author_list": "Lester, Henry A.; Lavis, Luke D.; et el." }, { "id": "https://authors.library.caltech.edu/records/gsawf-1aw27", "eprint_id": 57088, "eprint_status": "archive", "datestamp": "2023-08-22 16:25:07", "lastmod": "2023-10-23 17:02:40", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Post-Michael-R", "name": { "family": "Post", "given": "Michael R." }, "orcid": "0000-0002-3214-7619" }, { "id": "Limapichat-Walrati", "name": { "family": "Limapichat", "given": "Walrati" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Heterologous expression and nonsense suppression provide insights into agonist behavior at \u03b16\u03b22 nicotinic acetylcholine receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "Parkinson's disease; Addiction; Ion channels; Nicotinic acetylcholine receptors; Electrophysiology; Non-canonical amino acids", "note": "\u00a9 2015 Published by Elsevier Ltd.\n\nReceived 14 January 2015; Received in revised form 27 March 2015; Accepted 10 April 2015; Available online 20 April 2015.\n\nWe thank the NIH (NS 34407) for support of this work. MRP was supported by an NIH/NRSA training grant: 5 T32 GM07616.\n\nAccepted Version - nihms683389.pdf
Supplemental Material - mmc1.docx
Supplemental Material - mmc2.docx
", "abstract": "The \u03b16-containing subtypes of the nicotinic acetylcholine receptor (nAChR) are localized to presynaptic terminals of the dopaminergic pathways of the central nervous system. Selective ligands for these nAChRs are potentially useful in both Parkinson's disease and addiction. For these and other goals, it is important to distinguish the binding behavior of agonists at the \u03b16-\u03b22 binding site versus other subtypes. To study this problem, we apply nonsense suppression-based non-canonical amino acid mutagenesis. We report a combination of four mutations in \u03b16\u03b22 that yield high-level heterologous expression in Xenopus oocytes. By varying mRNA injection ratios, two populations were observed with unique characteristics, likely due to differing stoichiometries. Responses to nine known nAChR agonists were analyzed at the receptor, and their corresponding EC50 values and efficacies are reported. The system is compatible with nonsense suppression, allowing structure\u2013function studies between Trp149 \u2013 a conserved residue on loop B found to make a cation-\u03c0 interaction at several nAChR subtypes \u2013 and several agonists. These studies reveal that acetylcholine forms a strong cation-\u03c0 interaction with the conserved tryptophan, while nicotine and TC299423 do not, suggesting a unique pharmacology for the \u03b16\u03b22 nAChR.", "date": "2015-10", "date_type": "published", "publication": "Neuropharmacology", "volume": "97", "publisher": "Elsevier", "pagerange": "376-382", "id_number": "CaltechAUTHORS:20150429-110737552", "issn": "0028-3908", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150429-110737552", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5 T32 GM07616" } ] }, "doi": "10.1016/j.neuropharm.2015.04.009", "pmcid": "PMC4635625", "primary_object": { "basename": "mmc2.docx", "url": "https://authors.library.caltech.edu/records/gsawf-1aw27/files/mmc2.docx" }, "related_objects": [ { "basename": "nihms683389.pdf", "url": "https://authors.library.caltech.edu/records/gsawf-1aw27/files/nihms683389.pdf" }, { "basename": "mmc1.docx", "url": "https://authors.library.caltech.edu/records/gsawf-1aw27/files/mmc1.docx" } ], "resource_type": "article", "pub_year": "2015", "author_list": "Post, Michael R.; Limapichat, Walrati; et el." }, { "id": "https://authors.library.caltech.edu/records/4cjq5-cp673", "eprint_id": 54687, "eprint_status": "archive", "datestamp": "2023-08-22 16:12:48", "lastmod": "2023-10-20 20:06:58", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Inside-out neuropharmacology of nicotinic drugs", "ispublished": "pub", "full_text_status": "public", "keywords": "Chaperoning; Nicotine; Nicotine addiction; Nicotinic receptors; Unfolded protein response; Upregulation", "note": "\u00a9 2015 Published by Elsevier Ltd.\n\nAvailable online 4 February 2015.\n\nThis work was supported by National Institutes of Health Grants AG033954, DA017279, DA019375, DA030396, NS034407, and DA033721, by the California Tobacco-Related Disease Research Program Grant 17RT0127, and by Louis and Janet Fletcher. We thank Sherry Leonard for contributing comments.\n\nAccepted Version - nihms-665035.pdf
", "abstract": "Upregulation of neuronal nicotinic acetylcholine receptors (AChRs) is a venerable result of chronic exposure to nicotine; but it is one of several consequences of pharmacological chaperoning by nicotine and by some other nicotinic ligands, especially agonists. Nicotinic ligands permeate through cell membranes, bind to immature AChR oligomers, elicit incompletely understood conformational reorganizations, increase the interaction between adjacent AChR subunits, and enhance the maturation process toward stable AChR pentamers. These changes and stabilizations in turn lead to increases in both anterograde and retrograde traffic within the early secretory pathway. In addition to the eventual upregulation of AChRs at the plasma membrane, other effects of pharmacological chaperoning include modifications to endoplasmic reticulum stress and to the unfolded protein response. Because these processes depend on pharmacological chaperoning within intracellular organelles, we group them as \"inside-out pharmacology\". This term contrasts with the better-known, acute, \"outside-in\" effects of activating and desensitizing plasma membrane AChRs. We review current knowledge concerning the mechanisms and consequences of inside-out pharmacology.", "date": "2015-09", "date_type": "published", "publication": "Neuropharmacology", "volume": "96", "publisher": "Elsevier", "pagerange": "178-193", "id_number": "CaltechAUTHORS:20150211-075813471", "issn": "0028-3908", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150211-075813471", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "DA019375" }, { "agency": "NIH", "grant_number": "DA030396" }, { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "NIH", "grant_number": "DA033721" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "17RT0127" } ] }, "doi": "10.1016/j.neuropharm.2015.01.022", "pmcid": "PMC4486611", "primary_object": { "basename": "nihms-665035.pdf", "url": "https://authors.library.caltech.edu/records/4cjq5-cp673/files/nihms-665035.pdf" }, "resource_type": "article", "pub_year": "2015", "author_list": "Henderson, Brandon J. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/ekf2w-zha60", "eprint_id": 55159, "eprint_status": "archive", "datestamp": "2023-08-22 16:12:56", "lastmod": "2023-10-20 21:53:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Sinkus-M-L", "name": { "family": "Sinkus", "given": "Melissa L." } }, { "id": "Graw-S", "name": { "family": "Graw", "given": "Sharon" } }, { "id": "Freedman-R", "name": { "family": "Freedman", "given": "Robert" } }, { "id": "Ross-R-G", "name": { "family": "Ross", "given": "Randal G." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Leonard-S", "name": { "family": "Leonard", "given": "Sherry" } } ] }, "title": "The human CHRNA7 and CHRFAM7A genes: A review of the genetics, regulation, and function", "ispublished": "pub", "full_text_status": "public", "keywords": "Nicotinic receptor; Duplication; Schizophrenia; Alzheimer's; CHRNA7; CHRFAM7A; Gene mutation", "note": "\u00a9 2015 Published by Elsevier Ltd.\n\nAvailable online 19 February 2015.\n\nSpecial thanks to Ralph Berger, Judith Logel, Margaret Short, and William Proctor, Ph.D. for technical assistance. The research was funded by NIH grants DA09457, MH81177, and the Veterans Affairs Medical Research Service to SL.\n\nAccepted Version - nihms665644.pdf
", "abstract": "The human \u03b17 neuronal nicotinic acetylcholine receptor gene (CHRNA7) is ubiquitously expressed in both the central nervous system and in the periphery. CHRNA7 is genetically linked to multiple disorders with cognitive deficits, including schizophrenia, bipolar disorder, ADHD, epilepsy, Alzheimer's disease, and Rett syndrome. The regulation of CHRNA7 is complex; more than a dozen mechanisms are known, one of which is a partial duplication of the parent gene. Exons 5\u201310 of CHRNA7 on chromosome 15 were duplicated and inserted 1.6 Mb upstream of CHRNA7, interrupting an earlier partial duplication of two other genes. The chimeric CHRFAM7A gene product, dup\u03b17, assembles with \u03b17 subunits, resulting in a dominant negative regulation of function. The duplication is human specific, occurring neither in primates nor in rodents. The duplicated \u03b17 sequence in exons 5\u201310 of CHRFAM7A is almost identical to CHRNA7, and thus is not completely queried in high throughput genetic studies (GWAS). Further, pre-clinical animal models of the \u03b17nAChR utilized in drug development research do not have CHRFAM7A (dup\u03b17) and cannot fully model human drug responses. The wide expression of CHRNA7, its multiple functions and modes of regulation present challenges for study of this gene in disease.", "date": "2015-09", "date_type": "published", "publication": "Neuropharmacology", "volume": "96", "publisher": "Elsevier", "pagerange": "274-288", "id_number": "CaltechAUTHORS:20150224-151714505", "issn": "0028-3908", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150224-151714505", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA09457" }, { "agency": "NIH", "grant_number": "MH81177" }, { "agency": "Veterans Affairs Medical Research Service" } ] }, "doi": "10.1016/j.neuropharm.2015.02.006", "pmcid": "PMC4486515", "primary_object": { "basename": "nihms665644.pdf", "url": "https://authors.library.caltech.edu/records/ekf2w-zha60/files/nihms665644.pdf" }, "resource_type": "article", "pub_year": "2015", "author_list": "Sinkus, Melissa L.; Graw, Sharon; et el." }, { "id": "https://authors.library.caltech.edu/records/6z3r0-fva06", "eprint_id": 59025, "eprint_status": "archive", "datestamp": "2023-08-20 07:48:50", "lastmod": "2023-10-23 19:58:49", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Marotta-Christopher-B", "name": { "family": "Marotta", "given": "Christopher B." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "An Unaltered Orthosteric Site and a Network of Long-Range Allosteric Interactions for PNU-120596 in \u03b17 Nicotinic Acetylcholine Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2015 Elsevier Ltd. \n\nReceived: January 22, 2015; Revised: May 19, 2015; Accepted: June 10, 2015; Published: July 23, 2015. \n\nWe thank Matt Rienzo and Noah Duffy for their work in making the dCA-coupled fluorinated-OMe-tyrosines and tRNA, and Emily Blythe for developing the a7 homology model. Support for this work came from the NIH (NS 34407).\n\nAccepted Version - nihms708057.pdf
Supplemental Material - mmc1.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are vital to neuronal signaling, are implicated in important processes such as learning and memory, and are therapeutic targets for neural diseases. The \u03b17 nAChR has been implicated in Alzheimer's disease and schizophrenia, and allosteric modulators have become one focus of drug development efforts. We investigate the mode of action of the \u03b17-selective positive allosteric modulator, PNU-120596, and show that the higher potency of acetylcholine in the presence of PNU-120596 is not due to an altered agonist binding site. In addition, we propose several residues in the gating interface and transmembrane region that are functionally important to transduction of allosteric properties, and link PNU-120596, the acetylcholine binding region, and the receptor gate. These results suggest global protein stabilization from a communication network through several key residues that alter the gating equilibrium of the receptor while leaving the agonist binding properties unperturbed.", "date": "2015-08-20", "date_type": "published", "publication": "Chemistry and Biology", "volume": "22", "number": "8", "publisher": "Elsevier", "pagerange": "1063-1073", "id_number": "CaltechAUTHORS:20150728-085848506", "issn": "1074-5521", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150728-085848506", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" } ] }, "doi": "10.1016/j.chembiol.2015.06.018", "pmcid": "PMC4547686", "primary_object": { "basename": "mmc1.pdf", "url": "https://authors.library.caltech.edu/records/6z3r0-fva06/files/mmc1.pdf" }, "related_objects": [ { "basename": "nihms708057.pdf", "url": "https://authors.library.caltech.edu/records/6z3r0-fva06/files/nihms708057.pdf" } ], "resource_type": "article", "pub_year": "2015", "author_list": "Marotta, Christopher B.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/k1ryv-x5j45", "eprint_id": 57644, "eprint_status": "archive", "datestamp": "2023-08-20 06:16:10", "lastmod": "2023-10-23 17:35:56", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Wieskopf-Jeffrey-S", "name": { "family": "Wieskopf", "given": "Jeffrey S." } }, { "id": "Limapichat-Walrati", "name": { "family": "Limapichat", "given": "Walrati" } }, { "id": "Post-Michael-R", "name": { "family": "Post", "given": "Michael R." }, "orcid": "0000-0002-3214-7619" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "The nicotinic \u03b16 subunit gene determines variability in chronic pain sensitivity via cross-inhibition of P2X2/3 receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2015 American Association for the Advancement of Science.\n\nSubmitted 7 July 2014; accepted 11 March 2015; published 13 May 2015.\n\nWe thank T. Earley, T. Miyamoto, and M. Petrus for assistance with DRG dissection. We thank S. Ranade and V. Uzzell for data analysis. Funding: Supported by the Canadian Institutes for Health Research and the Louise and Alan Edwards Foundation (J.S.M.) and the NIH (D.A.D., H.A.L., and A.P.). Author contributions: The study was conceived by A.P. and J.S.M. and designed by J.S.W., L.D., I.B., M.I.D., H.A.L., A.P., and J.S.M. J.S.W., J. Mathur,\nW.L., M.R.P., M.A.-Q., R.E.S., L.J.M., K.F., J.-S.A., J.Z., J. Marcovitz, A.H.T., P.M.S., S.C., J.J., S.A.S., E.K.A., R.B., C.I.R., H.K., and J.W. collected data. D.V.Z., S.B.S., F.D., R.M.D., S.K.S., W.L., G.D.S., and A.I.S. analyzed data. U.M., J.-P.C., and M.D. provided reagents or data. W.M., L.D., I.B., D.A.D., S.C.R.L., M.I.D., H.A.L., A.P., and J.S.M. supervised the collection of data, contributed to its interpretation, and edited the manuscript. J.S.W. and J.S.M. wrote the manuscript. Competing interests: The authors declare that they have no competing interests. Data and materials availability: The expression genomics data for this study have been deposited into the GeneNetwork Database (www.genenetwork.org).\n\nAccepted Version - nihms814673.pdf
Supplemental Material - 7-287ra72_SM.pdf
", "abstract": "Chronic pain is a highly prevalent and poorly managed human health problem. We used microarray-based expression genomics in 25 inbred mouse strains to identify dorsal root ganglion (DRG)\u2013expressed genetic contributors to mechanical allodynia, a prominent symptom of chronic pain. We identified expression levels of Chrna6, which encodes the \u03b16 subunit of the nicotinic acetylcholine receptor (nAChR), as highly associated with allodynia. We confirmed the importance of \u03b16* (\u03b16-containing) nAChRs by analyzing both gain- and loss-of-function mutants. We find that mechanical allodynia associated with neuropathic and inflammatory injuries is significantly altered in \u03b16* mutants, and that \u03b16* but not \u03b14* nicotinic receptors are absolutely required for peripheral and/or spinal nicotine analgesia. Furthermore, we show that Chrna6's role in analgesia is at least partially due to direct interaction and cross-inhibition of \u03b16* nAChRs with P2X2/3 receptors in DRG nociceptors. Finally, we establish the relevance of our results to humans by the observation of genetic association in patients suffering from chronic postsurgical and temporomandibular pain.", "date": "2015-05-13", "date_type": "published", "publication": "Science Translational Medicine", "volume": "7", "number": "287", "publisher": "American Association for the Advancement of Science", "pagerange": "Art. No. 287ra72", "id_number": "CaltechAUTHORS:20150519-092649728", "issn": "1946-6234", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150519-092649728", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Canadian Institutes of Health Research (CIHR)" }, { "agency": "Louise and Alan Edwards Foundation" }, { "agency": "NIH" } ] }, "doi": "10.1126/scitranslmed.3009986", "pmcid": "PMC5018401", "primary_object": { "basename": "7-287ra72_SM.pdf", "url": "https://authors.library.caltech.edu/records/k1ryv-x5j45/files/7-287ra72_SM.pdf" }, "related_objects": [ { "basename": "nihms814673.pdf", "url": "https://authors.library.caltech.edu/records/k1ryv-x5j45/files/nihms814673.pdf" } ], "resource_type": "article", "pub_year": "2015", "author_list": "Wieskopf, Jeffrey S.; Limapichat, Walrati; et el." }, { "id": "https://authors.library.caltech.edu/records/pe26h-xz928", "eprint_id": 53197, "eprint_status": "archive", "datestamp": "2023-08-22 15:14:35", "lastmod": "2023-10-19 14:38:44", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Miles-T-F", "name": { "family": "Miles", "given": "Timothy F." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Allosteric activation of the 5-HT_3AB receptor by mCPBG", "ispublished": "pub", "full_text_status": "public", "keywords": "mCPBG; Serotonin; 5-HT3; Allosteric modulation; Cys-loop receptor", "note": "\u00a9 2014 Published by Elsevier Ltd. \n\nReceived 8 July 2014. Received in revised form 16 October 2014. Accepted 10 December 2014. Available online 23 December 2014. \n\nWe would like to thank Dr. Sarah Lummis (Cambridge) and Dr.\nNoah Duffy for helpful discussion. \n\nThis work was supported by the NIH (NS 34407).\n\nAccepted Version - nihms651345.pdf
", "abstract": "The 5-HT_3AB receptor contains three A and two B subunits in an A-A-B-A-B order. However, serotonin function at the 5-HT_3AB receptor has been shown to depend solely on the A-A interface present in the homomeric receptor. Using mutations at sites on both the primary (E122) and complementary (Y146) faces of the B subunit, we demonstrate that meta-chlorophenyl biguanide (mCPBG), a 5-HT_3 selective agonist, is capable of binding to and activating the 5-HT_3AB receptor at all five subunit interfaces of the heteromer. Further, mCPBG is capable of allosterically modulating the activity of serotonin from these sites. While these five binding sites are similar enough that they form to a monophasic dose \u2013 response relationship, we uncover subtle differences in the heteromeric binding sites. We also find that the A-A interface appears to contribute disproportionately to the efficacy of 5-HT_3AB receptor activation.", "date": "2015-04", "date_type": "published", "publication": "Neuropharmacology", "volume": "91", "publisher": "Elsevier", "pagerange": "103-108", "id_number": "CaltechAUTHORS:20150106-073218861", "issn": "0028-3908", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150106-073218861", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" } ] }, "doi": "10.1016/j.neuropharm.2014.12.018", "pmcid": "PMC4312754", "primary_object": { "basename": "nihms651345.pdf", "url": "https://authors.library.caltech.edu/records/pe26h-xz928/files/nihms651345.pdf" }, "resource_type": "article", "pub_year": "2015", "author_list": "Miles, Timothy F.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/2mqqd-98s92", "eprint_id": 55653, "eprint_status": "archive", "datestamp": "2023-08-22 15:08:01", "lastmod": "2023-10-20 22:55:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Miwa-Julie-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Wang-Ying", "name": { "family": "Wang", "given": "Ying" } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Nicotinic Receptor Subtype-Selective Circuit Patterns in the Subthalamic Nucleus", "ispublished": "pub", "full_text_status": "public", "keywords": "alpha4beta2; alpha7; chronic nicotine; Parkinson's disease; substantia nigra; upregulation", "note": "\u00a9 2015 the authors. After 6 months the work becomes available to the public to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license. \n\nReceived Aug. 17, 2014; revised Jan. 5, 2015; accepted Jan. 14, 2015.\n\nThis work was supported by National Institutes of Health Grants DA17279, AG033954, and R21DA033831, and the California Tobacco-Related Disease Research Program Grants 16FT-0066, 17RT-0127, and 19KT-0032.We thank Raad Nashmi and Haijiang Cai for comments and discussion.\n\nPublished - 3734.full.pdf
", "abstract": "The glutamatergic subthalamic nucleus (STN) exerts control over motor output through nuclei of the basal ganglia. High-frequency electrical stimuli in the STN effectively alleviate motor symptoms in movement disorders, and cholinergic stimulation boosts this effect. To gain knowledge about the mechanisms of cholinergic modulation in the STN, we studied cellular and circuit aspects of nicotinic acetylcholine receptors (nAChRs) in mouse STN. We discovered two largely divergent microcircuits in the STN; these are regulated in part by either \u03b14\u03b22 or \u03b17 nAChRs. STN neurons containing \u03b14\u03b22 nAChRs (\u03b14\u03b22 neurons) received more glutamatergic inputs, and preferentially innervated GABAergic neurons in the substantia nigra pars reticulata. In contrast, STN neurons containing \u03b17 nAChRs (\u03b17 neurons) received more GABAergic inputs, and preferentially innervated dopaminergic neurons in the substantia nigra pars compacta. Interestingly, local electrical stimuli excited a majority (79%) of \u03b14\u03b22 neurons but exerted strong inhibition in 58% of \u03b17 neurons, indicating an additional diversity of STN neurons: responses to electrical stimulation. Chronic exposure to nicotine selectively affects \u03b14\u03b22 nAChRs in STN: this treatment increased the number of \u03b14\u03b22 neurons, upregulated \u03b14-containing nAChR number and sensitivity, and enhanced the basal firing rate of \u03b14\u03b22 neurons both ex vivo and in vivo. Thus, chronic nicotine enhances the function of the microcircuit involving \u03b14\u03b22 nAChRs. This indicates chronic exposure to nicotinic agonist as a potential pharmacological intervention to alter selectively the balance between these two microcircuits, and may provide a means to inhibit substantia nigra dopaminergic neurons.", "date": "2015-03-04", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "35", "number": "9", "publisher": "Society for Neuroscience", "pagerange": "3734-3746", "id_number": "CaltechAUTHORS:20150309-153924379", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150309-153924379", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "R21DA033831" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "16FT-0066" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "17RT-0127" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19KT-0032" } ] }, "doi": "10.1523/JNEUROSCI.3528-14.2015", "pmcid": "PMC4348180", "primary_object": { "basename": "3734.full.pdf", "url": "https://authors.library.caltech.edu/records/2mqqd-98s92/files/3734.full.pdf" }, "resource_type": "article", "pub_year": "2015", "author_list": "Xiao, Cheng; Miwa, Julie M.; et el." }, { "id": "https://authors.library.caltech.edu/records/9c6rn-2cj64", "eprint_id": 49689, "eprint_status": "archive", "datestamp": "2023-08-22 14:09:41", "lastmod": "2023-10-17 22:05:04", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nichols-W-A", "name": { "family": "Nichols", "given": "Weston A." } }, { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Yu-Caroline", "name": { "family": "Yu", "given": "Caroline" } }, { "id": "Parker-R-L", "name": { "family": "Parker", "given": "Rell L." } }, { "id": "Richards-C-I", "name": { "family": "Richards", "given": "Christopher I." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Miwa-J-M", "name": { "family": "Miwa", "given": "Julie M." } } ] }, "title": "Lynx1 Shifts \u03b14\u03b22 Nicotinic Receptor Subunit Stoichiometry by Affecting Assembly in the Endoplasmic Reticulum", "ispublished": "pub", "full_text_status": "public", "keywords": "Cys-loop receptor; glycosylphosphatidylinositol (GPI anchor); nicotinic acetylcholine receptors (nAChR); snake venom; toxin", "note": "\u00a9 2014 The American Society for Biochemistry and Molecular Biology.\n\nReceived April 23, 2014; accepted September 5, 2014.\n\nThanks to Sheri McKinney for providing hippocampal neuron cultures. We also thank R. Srinivasan and M. Starbird for comments and technical advice. This research was supported by TRDRP 19KT-0032 and by the National Institutes of Health (DA033831, NS034407, EY018502, MH088550).\n\nPublished - 31423.full.pdf
", "abstract": "GPI-anchored neurotoxin-like receptor binding proteins, such as lynx modulators, are topologically positioned to exert pharmacological effects by binding to the extracellular portion of nAChRs. These actions are generally thought to proceed when both lynx and the nAChRs are on the plasma membrane. Here, we demonstrate that lynx1 also exerts effects on \u03b14\u03b22 nAChRs within the endoplasmic reticulum. Lynx1 affects assembly of nascent \u03b14 and \u03b22 subunits, and alters the stoichiometry of the population that reaches the plasma membrane. Additionally, these data suggest that lynx1 shifts nAChR stoichiometry to low sensitivity (\u03b14)_3 (\u03b22)_2 pentamers primarily through this interaction in the endoplasmic reticulum, rather than solely via direct modulation of activity on the plasma membrane To our knowledge, these data represent the first test of the hypothesis that a lynx family member, or indeed any GPI-anchored protein, could act within the cell to alter assembly of multi-subunit protein.", "date": "2014-11-07", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "289", "number": "45", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "31423-31432", "id_number": "CaltechAUTHORS:20140915-092705574", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140915-092705574", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19KT-0032" }, { "agency": "NIH", "grant_number": "DA033831" }, { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "NIH", "grant_number": "EY018502" }, { "agency": "NIH", "grant_number": "MH088550" } ] }, "doi": "10.1074/jbc.M114.573667", "pmcid": "PMC4223341", "primary_object": { "basename": "31423.full.pdf", "url": "https://authors.library.caltech.edu/records/9c6rn-2cj64/files/31423.full.pdf" }, "resource_type": "article", "pub_year": "2014", "author_list": "Nichols, Weston A.; Henderson, Brandon J.; et el." }, { "id": "https://authors.library.caltech.edu/records/4pp35-rsg18", "eprint_id": 48319, "eprint_status": "archive", "datestamp": "2023-08-20 03:03:48", "lastmod": "2023-10-17 18:42:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Daeffler-K-N-M", "name": { "family": "Daeffler", "given": "Kristina N.-M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Functional Evaluation of Key Interactions Evident in the Structure of the Eukaryotic Cys-Loop Receptor GluCl", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2014 American Chemical Society. ACS AuthorChoice - This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes. \n\nReceived: April 29, 2014; accepted: July 22, 2014; Published: July 22, 2014. \n\nThis work was supported by the National Institutes of Health\ngrant NS034407. We are also grateful for support from the\nBeckman Institute Zebrafish/Xenopus/Lamprey Facility.\n\nPublished - cb500323d.pdf
Supplemental Material - cb500323d_si_001.pdf
", "abstract": "The publication of the first high-resolution crystal structure of a eukaryotic Cys-loop receptor, GluCl\u03b1, has provided valuable structural information on this important class of ligand-gated ion channels (LGIC). However, limited functional data exist for the GluCl receptors. Before applying the structural insights from GluCl to mammalian Cys-loop receptors such as nicotinic acetylcholine and GABA receptors, it is important to ensure that established functional features of mammalian Cys-loop receptors are present in the more distantly related GluCl receptors. Here, we seek to identify ligand-binding interactions that are generally associated with Cys-loop receptors, including the frequently observed cation\u2212\u03c0 interaction. Our studies were performed on the highly homologous GluCl\u03b2 receptor, because GluCl\u03b1 is not activated by glutamate in Xenopus laevis oocytes. Mutagenesis of the signal peptide and pore lining helix was performed to enhance functional expression and sensitivity to applied ligand, respectively. Conventional and unnatural amino acid mutagenesis indicate a strong cation\u2212\u03c0 interaction between Y206 and the protonated amine of glutamate, as well as other important ionic and hydrogen bond interactions between the ligand and the binding site, consistent with the crystal structure.", "date": "2014-10", "date_type": "published", "publication": "ACS Chemical Biology", "volume": "9", "number": "10", "publisher": "American Chemical Society", "pagerange": "2283-2290", "id_number": "CaltechAUTHORS:20140811-132238553", "issn": "1554-8929", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140811-132238553", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "Caltech Beckman Institute" } ] }, "doi": "10.1021/cb500323d", "pmcid": "PMC4201344", "primary_object": { "basename": "cb500323d.pdf", "url": "https://authors.library.caltech.edu/records/4pp35-rsg18/files/cb500323d.pdf" }, "related_objects": [ { "basename": "cb500323d_si_001.pdf", "url": "https://authors.library.caltech.edu/records/4pp35-rsg18/files/cb500323d_si_001.pdf" } ], "resource_type": "article", "pub_year": "2014", "author_list": "Daeffler, Kristina N.-M.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/ea6d4-g9206", "eprint_id": 47546, "eprint_status": "archive", "datestamp": "2023-08-22 13:41:35", "lastmod": "2023-10-26 20:41:46", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Wang-Ying", "name": { "family": "Wang", "given": "Ying" } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Indersmitten-T", "name": { "family": "Indersmitten", "given": "Tim" } }, { "id": "Freedman-R", "name": { "family": "Freedman", "given": "Robert" } }, { "id": "Leonard-S", "name": { "family": "Leonard", "given": "Sherry" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "The duplicated \u03b17 subunits assemble and form functional nicotinic receptors with the full-length \u03b17", "ispublished": "pub", "full_text_status": "public", "keywords": "electrophysiology; fluorescence recovery after photobleaching (FRAP); fluorescence resonance energy transfer (FRET); genomics; ion channel; nicotinic acetylcholine receptors (nAChR); patch clamp; schizophrenia; choline; ligand-gated channel", "note": "\u00a9 2014 The American Society for Biochemistry and Molecular Biology.\n\nPublished on July 23, 2014 as Manuscript M114.582858.\n\nThanks to Sheri McKinney for providing neuron cultures. We also thank Drs. Christopher I. Richards and Bruce N. Cohen for helpful discussions.\n\nThis work was supported by the National Institutes of Health (MH088550).\n\nPublished - J._Biol._Chem.-2014-Wang-26451-63.pdf
", "abstract": "The \u03b17 nicotinic acetylcholine receptor gene (CHRNA7) is linked to schizophrenia. A partial duplication of CHRNA7 (CHRFAM7A) is found in humans on 15q13-14. Exon 6 of CHRFAM7A harbors a 2 base pair deletion polymorphism, CHRFAM7A\u03942bp, which is also associated with schizophrenia. To understand the effects of the duplicated subunits on \u03b17 receptors, we fused \u03b17, dup\u03b17, and dup\u0394\u03b17 subunits with various fluorescent proteins. The duplicated subunits co-localized with full-length \u03b17 subunits in mouse neuroblastoma cells (Neuro2a) as well as rat hippocampal neurons. We investigated the interaction between the duplicated subunits and full-length \u03b17 by measuring Foerster resonance energy transfer (FRET) using donor recovery after photobleaching (DRAP) and fluorescence lifetime imaging microscopy (FLIM). The results revealed that the duplicated proteins co-assemble with \u03b17. In electrophysiological studies, leucine at the 9' position in the M2 membrane-spanning segment was replaced with Cys in dup\u03b17 or dup\u0394\u03b17, and constructs were cotransfected with full-length \u03b17 in Neuro2a cells. Exposure to ethylammonium methanethiosulfonate (MTSEA) inhibited acetylcholine (ACh)-induced currents, showing that the assembled functional nAChRs included the duplicated subunit. Incorporation of dup\u03b17 and dup\u2206\u03b17 subunits modestly changes the sensitivity of receptors to choline and varenicline. Thus, the duplicated proteins are assembled and transported to the cell membrane together with full-length \u03b17 subunits, and alter the function of the nAChRs. The characterization of dup\u03b17 and dup\u0394\u03b17 as well as their influence on \u03b17 nAChRs may help explain the pathophysiology of schizophrenia and may suggest therapeutic strategies.", "date": "2014-09-19", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "289", "number": "38", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "26451-26463", "id_number": "CaltechAUTHORS:20140729-082327052", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140729-082327052", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "MH088550" } ] }, "doi": "10.1074/jbc.M114.582858", "pmcid": "PMC4176222", "primary_object": { "basename": "J._Biol._Chem.-2014-Wang-26451-63.pdf", "url": "https://authors.library.caltech.edu/records/ea6d4-g9206/files/J._Biol._Chem.-2014-Wang-26451-63.pdf" }, "resource_type": "article", "pub_year": "2014", "author_list": "Wang, Ying; Xiao, Cheng; et el." }, { "id": "https://authors.library.caltech.edu/records/tsvpp-1q848", "eprint_id": 50432, "eprint_status": "archive", "datestamp": "2023-08-20 02:44:56", "lastmod": "2023-10-17 23:48:23", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Limapichat-W", "name": { "family": "Limapichat", "given": "Walrati" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Subtype-Specific Mechanisms for Functional Interaction between \u03b16\u03b24* Nicotinic Acetylcholine Receptors and P2X Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2014 by The American Society for Pharmacology and Experimental Therapeutics.\n\nReceived April 11, 2014; accepted June 24, 2014.\n\nThe authors thank J. Mogil, J. Wieskopf, R. Drenan, M. AlQazzaz, and C. I. Richards for discussion.\n\nSupplemental Material - 93179_2014-06-15_supplementary_info.pdf
", "abstract": "P2X receptors and nicotinic acetylcholine receptors (nAChRs) display functional and physical interactions in many cell types and heterologous expression systems, but interactions between \u03b16\u03b24-containing (\u03b16\u03b24*) nAChRs and P2X2 receptors and/or P2X3 receptors have not been fully characterized. We measured several types of crosstalk in oocytes coexpressing \u03b16\u03b24 nAChRs and P2X2, P2X3, or P2X2/3 receptors. A novel form of crosstalk occurs between \u03b16\u03b24 nAChRs and P2X2 receptors. P2X2 receptors were forced into a prolonged desensitized state upon activation by ATP through a mechanism that does not depend on the intracellular C terminus of the P2X2 receptors. Coexpression of \u03b16\u03b24 nAChRs with P2X3 receptors shifts the ATP dose-response relation to the right, even in the absence of acetylcholine (ACh). Moreover, currents become nonadditive when ACh and ATP are coapplied, as previously reported for other Cys-loop receptors interacting with P2X receptors, and this crosstalk is dependent on the presence of the P2X3 C-terminal domain. P2X2 receptors also functionally interact with \u03b16\u03b24\u03b23 but through a different mechanism from \u03b16\u03b24. The interaction with P2X3 receptors is less pronounced for the \u03b16\u03b24\u03b23 nAChR than the \u03b16\u03b24 nAChR. We also measured a functional interaction between the \u03b16\u03b24 nAChRs and the heteromeric P2X2/3 receptor. Experiments with the nAChR channel blocker mecamylamine on P2X2\u2013\u03b16\u03b24 oocytes point to the loss of P2X2 channel activity during the crosstalk, whereas the ion channel pores of the P2X receptors were fully functional and unaltered by the receptor interaction for P2X2\u2013\u03b16\u03b24\u03b23, P2X2/3\u2013\u03b16\u03b24, and P2X2/3\u2013\u03b16\u03b24\u03b23. These results may be relevant to dorsal root ganglion cells and to other neurons that coexpress these receptor subunits.", "date": "2014-09", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "86", "number": "3", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "263-274", "id_number": "CaltechAUTHORS:20141016-090934738", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20141016-090934738", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1124/mol.114.093179", "pmcid": "PMC4152909", "primary_object": { "basename": "93179_2014-06-15_supplementary_info.pdf", "url": "https://authors.library.caltech.edu/records/tsvpp-1q848/files/93179_2014-06-15_supplementary_info.pdf" }, "resource_type": "article", "pub_year": "2014", "author_list": "Limapichat, Walrati; Dougherty, Dennis A.; et el." }, { "id": "https://authors.library.caltech.edu/records/vjr3n-k2362", "eprint_id": 47358, "eprint_status": "archive", "datestamp": "2023-08-22 13:12:39", "lastmod": "2023-10-26 20:31:30", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kobayashi-Atsuko-K", "name": { "family": "Kobayashi", "given": "Atsuko" } }, { "id": "Parker-R-L", "name": { "family": "Parker", "given": "Rell L." } }, { "id": "Wright-A-P", "name": { "family": "Wright", "given": "Ashley P." } }, { "id": "Brahem-H", "name": { "family": "Brahem", "given": "Hajer" } }, { "id": "Ku-Pauline", "name": { "family": "Ku", "given": "Pauline" } }, { "id": "Oliver-K-M", "name": { "family": "Oliver", "given": "Katherine M." } }, { "id": "Walz-A", "name": { "family": "Walz", "given": "Andreas" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Miwa-J-M", "name": { "family": "Miwa", "given": "Julie M." } } ] }, "title": "lynx1 Supports Neuronal Health in the Mouse Dorsal Striatum During Aging: an Ultrastructural Investigation", "ispublished": "pub", "full_text_status": "public", "keywords": "Nicotinic acetylcholine receptors; Cholinergic system; Prototoxins; Neurotoxins; Neurodegeneration; Plasticity; Learning and memory", "note": "\u00a9 2014 Springer Science+Business Media New York. \n\nReceived: 30 August 2013; accepted: 9 June 2014. \n\nThis work was supported by funds from Tobacco-Related Disease Research Program of the University of California, Grant Number TRDRP19KT-0032 for JMM and RLP, TRDRP22DT-0008 and NIH/NRSA Institutional training grant 5 T32 GM07616 for RLP; US- India BRCP Award - 1R21DA033831 for JMM; R01AG-033954 for HAL and JMM; R41DA032464 and 1R43MH094004 for PK, HB, APW and AW. Financial support for this project included funds for undergraduate research from the College of Arts & Sciences and the Department of Biological Sciences at Lehigh University for KMO. The Jensen electron microscopy facility is funded in part by the Gordon and Betty Moore Foundation, the Agouron Institute and the Beckman Foundation. Special thanks to Dr. Amber Rice for helpful discussion and critical reading of the manuscript, and to Samantha Eichelberger for editorial help. In memory of Andreas Walz.\n\nAccepted Version - nihms614070.pdf
", "abstract": "Nicotinic acetylcholine receptors have been shown to participate in neuroprotection in the aging brain. Lynx protein modulators dampen the activity of the cholinergic system through direct interaction with nicotinic receptors. Although lynx1 null mutant mice exhibit augmented learning and plasticity, they also exhibit macroscopic vacuolation in the dorsal striatum as they age, detectable at the optical microscope level. Despite the relevance of the lynx1 gene to brain function, little is known about the cellular ultrastructure of these age-related changes. In this study, we assessed degeneration in the dorsal striatum in 1-, 3-, 7-, and 13-month-old mice, using optical and transmission electron microscopy. We observed a loss of nerve fibers, a breakdown in nerve fiber bundles, and a loss of neuronal nuclei in the 13-month-old lynx1 null striatum. At higher magnification, these nerve fibers displayed intracellular vacuoles and disordered myelin sheaths. Few or none of these morphological alterations were present in younger lynx1 null mutant mice or in heterozygous lynx1 null mutant mice at any age. These data indicate that neuronal health can be maintained by titrating lynx1 dosage and that the lynx1 gene may participate in a trade-off between neuroprotection and augmented learning.", "date": "2014-07-17", "date_type": "published", "publication": "Journal of Molecular Neuroscience", "volume": "53", "number": "3", "publisher": "Humana Press", "pagerange": "525-536", "id_number": "CaltechAUTHORS:20140721-090832827", "issn": "0895-8696", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140721-090832827", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "TRDRP19KT-0032" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "TRDRP22DT-0008" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5T32 GM07616" }, { "agency": "NIH", "grant_number": "1R21DA033831" }, { "agency": "NIH", "grant_number": "R01AG-033954" }, { "agency": "NIH", "grant_number": "R41DA032464" }, { "agency": "NIH", "grant_number": "1R43MH094004" }, { "agency": "Lehigh University" }, { "agency": "Gordon and Betty Moore Foundation" }, { "agency": "Agouron Institute" }, { "agency": "Arnold and Mabel Beckman Foundation" }, { "agency": "US-India Bilateral Brain Research Collaborative Partnerships" } ] }, "doi": "10.1007/s12031-014-0352-1", "pmcid": "PMC4265479", "primary_object": { "basename": "nihms614070.pdf", "url": "https://authors.library.caltech.edu/records/vjr3n-k2362/files/nihms614070.pdf" }, "resource_type": "article", "pub_year": "2014", "author_list": "Kobayashi, Atsuko; Parker, Rell L.; et el." }, { "id": "https://authors.library.caltech.edu/records/h0555-yha70", "eprint_id": 48559, "eprint_status": "archive", "datestamp": "2023-08-22 13:12:29", "lastmod": "2023-10-17 19:26:34", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Shih-Pei-Yu", "name": { "family": "Shih", "given": "Pei-Yu" } }, { "id": "Engle-S-E", "name": { "family": "Engle", "given": "Staci E." } }, { "id": "Oh-Gyeon", "name": { "family": "Oh", "given": "Gyeon" } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Puskar-N-L", "name": { "family": "Puskar", "given": "Nyssa L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" } ] }, "title": "Differential Expression and Function of Nicotinic Acetylcholine Receptors in Subdivisions of Medial Habenula", "ispublished": "pub", "full_text_status": "public", "keywords": "habenula; interpeduncular; nicotine; nicotinic; tobacco; withdrawal", "note": "\u00a9 2014 The Authors. The authors grant the Society for Neuroscience an exclusive license to publish their work for the first 6 months. After 6 months the work becomes available to the public to copy, distribute, or display under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license.\n\nReceived Jan. 31, 2014; revised June 9, 2014; accepted June 12, 2014.\n\nThis work was supported by a Brain and Behavior Research Foundation (formerly National Alliance for Research on Schizophrenia and Depression) Young Investigator Award and National Institutes of Health Grant DA030396 to R.M.D., Grant DA028955 to H.A.L. and Michael J. Marks, and Grant NS034407 to Dennis A. Dougherty). We thank Elisha Mackey and Rahul Srinivasan for technical assistance.\n\nPublished - 9789.full.pdf
", "abstract": "Neuronal nAChRs in the medial habenula (MHb) to the interpeduncular nucleus (IPN) pathway are key mediators of nicotine's aversive properties. In this paper, we report new details regarding nAChR anatomical localization and function in MHb and IPN. A new group of knock-in mice were created that each expresses a single nAChR subunit fused to GFP, allowing high-resolution mapping. We find that \u03b13 and \u03b24 nAChR subunit levels are strong throughout the ventral MHb (MHbV). In contrast, \u03b16, \u03b22, \u03b23, and \u03b14 subunits are selectively found in some, but not all, areas of MHbV. All subunits were found in both ChAT-positive and ChAT-negative cells in MHbV. Next, we examined functional properties of neurons in the lateral and central part of MHbV (MHbVL and MHbVC) using brain slice patch-clamp recordings. MHbVL neurons were more excitable than MHbVC neurons, and they also responded more strongly to puffs of nicotine. In addition, we studied firing responses of MHbVL and MHbVC neurons in response to bath-applied nicotine. Cells in MHbVL, but not those in MHbVC, increased their firing substantially in response to 1 \u03bcm nicotine. Additionally, MHbVL neurons from mice that underwent withdrawal from chronic nicotine were less responsive to nicotine application compared with mice withdrawn from chronic saline. Last, we characterized rostral and dorsomedial IPN neurons that receive input from MHbVL axons. Together, our data provide new details regarding neurophysiology and nAChR localization and function in cells within the MHbV.", "date": "2014-07-16", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "34", "number": "29", "publisher": "Society for Neuroscience", "pagerange": "9789-9802", "id_number": "CaltechAUTHORS:20140814-112513666", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140814-112513666", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Brain and Behavior Research Foundation" }, { "agency": "NIH", "grant_number": "DA030396" }, { "agency": "NIH", "grant_number": "DA028955" }, { "agency": "NIH", "grant_number": "NS034407" } ] }, "doi": "10.1523/JNEUROSCI.0476-14.2014", "pmcid": "PMC4099552", "primary_object": { "basename": "9789.full.pdf", "url": "https://authors.library.caltech.edu/records/h0555-yha70/files/9789.full.pdf" }, "resource_type": "article", "pub_year": "2014", "author_list": "Shih, Pei-Yu; Engle, Staci E.; et el." }, { "id": "https://authors.library.caltech.edu/records/m0svv-4js11", "eprint_id": 74673, "eprint_status": "archive", "datestamp": "2023-08-20 00:58:57", "lastmod": "2023-10-24 22:50:54", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Marotta-C-B", "name": { "family": "Marotta", "given": "Christopher B." } }, { "id": "Rreza-I", "name": { "family": "Rreza", "given": "Iva" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Selective Ligand Behaviors Provide New Insights into Agonist Activation of Nicotinic Acetylcholine Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2014 American Chemical Society. This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes. \n\nReceived: December 20, 2013; Accepted: February 24, 2014; Published: February 24, 2014. \n\nWe thank the National Institutes of Health (NIH) (NS034407, DA017279, DA280382), NIH/NRSA (GM07616), and the California Tobacco-Related Disease Research Program from the University of California (19XT-0102) for support of this work.\n\nThe authors declare no competing financial interest.\n\nPublished - cb400937d.pdf
Supplemental Material - cb400937d_si_001.pdf
", "abstract": "Nicotinic acetylcholine receptors are a diverse set of ion channels that are essential to everyday brain function. Contemporary research studies selective activation of individual subtypes of receptors, with the hope of increasing our understanding of behavioral responses and neurodegenerative diseases. Here, we aim to expand current binding models to help explain the specificity seen among three activators of \u03b14\u03b22 receptors: sazetidine-A, cytisine, and NS9283. Through mutational analysis, we can interchange the activation profiles of the stoichiometry-selective compounds sazetidine-A and cytisine. In addition, mutations render NS9283\u2014currently identified as a positive allosteric modulator\u2014into an agonist. These results lead to two conclusions: (1) occupation at each primary face of an \u03b1 subunit is needed to activate the channel and (2) the complementary face of the adjacent subunit dictates the binding ability of the agonist.", "date": "2014-05-16", "date_type": "published", "publication": "ACS Chemical Biology", "volume": "9", "number": "5", "publisher": "American Chemical Society", "pagerange": "1153-1159", "id_number": "CaltechAUTHORS:20170302-160217090", "issn": "1554-8929", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170302-160217090", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "DA280382" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM07616" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19XT-0102" } ] }, "doi": "10.1021/cb400937d", "pmcid": "PMC4033646", "primary_object": { "basename": "cb400937d_si_001.pdf", "url": "https://authors.library.caltech.edu/records/m0svv-4js11/files/cb400937d_si_001.pdf" }, "related_objects": [ { "basename": "cb400937d.pdf", "url": "https://authors.library.caltech.edu/records/m0svv-4js11/files/cb400937d.pdf" } ], "resource_type": "article", "pub_year": "2014", "author_list": "Marotta, Christopher B.; Rreza, Iva; et el." }, { "id": "https://authors.library.caltech.edu/records/19vwb-sa496", "eprint_id": 47478, "eprint_status": "archive", "datestamp": "2023-08-22 12:36:42", "lastmod": "2023-10-26 20:38:21", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Richards-C-I", "name": { "family": "Richards", "given": "Christopher I." } } ] }, "title": "Pharmacological chaperoning of nAChRs: A therapeutic target for Parkinson's disease", "ispublished": "pub", "full_text_status": "public", "keywords": "Pharmacological chaperone; Chaperoning; Nicotine; nAChR; Tobacco; Neuroprotection; Parkinson's disease; Neurodegeneration; Unfolded protein response; Dopaminergic; Endoplasmic reticulum stress; FRET; TIRF; Confocal; ER exit sites; Golgi; Ligand; COPII; COPI", "note": "\u00a9 2014 Elsevier Ltd. \n\nReceived 31 December 2013; Received in revised form 18 February 2014; Accepted 18 February 2014. \n\nSupported by grants from the Tobacco-Related Disease Research Program (TRDRP 18FT-0066), the Michael J Fox Foundation (MJFF), U.S. National Institutes of Health, Louis and Janet Fletcher. \n\nThe authors declare no competing financial interests.\n\nAccepted Version - nihms-947291.pdf
", "abstract": "Chronic exposure to nicotine results in an upregulation of neuronal nicotinic acetylcholine receptors (nAChRs) at the cellular plasma membrane. nAChR upregulation occurs via nicotine-mediated pharmacological receptor chaperoning and is thought to contribute to the addictive properties of tobacco as well as relapse following smoking cessation. At the subcellular level, pharmacological chaperoning by nicotine and nicotinic ligands causes profound changes in the structure and function of the endoplasmic reticulum (ER), ER exit sites, the Golgi apparatus and secretory vesicles of cells. Chaperoning-induced changes in cell physiology exert an overall inhibitory effect on the ER stress/unfolded protein response. Cell autonomous factors such as the repertoire of nAChR subtypes expressed by neurons and the pharmacological properties of nicotinic ligands (full or partial agonist versus competitive antagonist) govern the efficiency of receptor chaperoning and upregulation. Together, these findings are beginning to pave the way for developing pharmacological chaperones to treat Parkinson's disease and nicotine addiction.", "date": "2014-05", "date_type": "published", "publication": "Pharmacological Research", "volume": "83", "publisher": "Elsevier", "pagerange": "20-29", "id_number": "CaltechAUTHORS:20140724-150050094", "issn": "1043-6618", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140724-150050094", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "18FT-0066" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "NIH" }, { "agency": "Louis and Janet Fletcher" } ] }, "doi": "10.1016/j.phrs.2014.02.005", "pmcid": "PMC6075820", "primary_object": { "basename": "nihms-947291.pdf", "url": "https://authors.library.caltech.edu/records/19vwb-sa496/files/nihms-947291.pdf" }, "resource_type": "article", "pub_year": "2014", "author_list": "Srinivasan, Rahul; Henderson, Brandon J.; et el." }, { "id": "https://authors.library.caltech.edu/records/zjk2e-yk085", "eprint_id": 54427, "eprint_status": "archive", "datestamp": "2023-08-20 00:16:22", "lastmod": "2023-10-20 16:26:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry" }, "orcid": "0000-0002-5470-5255" }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce" } } ] }, "title": "Drugs and the brain, the MOOC", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2014 Federation of American Societies for Experimental Biology.\n\nPublished - Lester_2014p221.2.pdf
", "abstract": "How does a MOOC differ in value and impact from previous \"distance-learning\" technologies? (1) A major additional possibility is the online community that springs up around the course. \"Drugs and the Brain\" brought some people with real-life interest in drug mechanisms, and others with detailed scientific experience. Quantifying the letter group: of the students who finished the 2012-2013 course and earned \"statements of accomplishment\", ~ 10% of the students had PHD degrees and 7% had MD degrees, and ~ 7% had professional degrees in pharmacy. This online community requires detailed curation, moderation, and judgment. For the 2013-2014 rendition, we plan to have two undergraduate TA's as curators. (2) A secondary possibility is the concept of instantaneous feedback on exams and quizzes. This can also be incorporated in previous technologies. What are limitations of the MOOC? (1) After all limitations in bandwidth and technology have been overcome, \"internet time\" will still be constrained by the fact that the world is round. The instructor will be asleep when some students want to communicate, and vice-versa. (2) The course staff must invest formidable effort to optimize the learning experience. In this area, we include analyzing quiz responses, clicks, etc. We also include human evaluation of essays and other student responses. These are both wonderful problems, in the sense that we have too much data!", "date": "2014-04", "date_type": "published", "publication": "FASEB Journal", "volume": "28", "number": "1", "publisher": "Federation of American Societies for Experimental Biology", "pagerange": "221.2", "id_number": "CaltechAUTHORS:20150205-130837428", "issn": "0892-6638", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150205-130837428", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "primary_object": { "basename": "Lester_2014p221.2.pdf", "url": "https://authors.library.caltech.edu/records/zjk2e-yk085/files/Lester_2014p221.2.pdf" }, "resource_type": "article", "pub_year": "2014", "author_list": "Lester, Henry and Cohen, Bruce" }, { "id": "https://authors.library.caltech.edu/records/501sm-h7s17", "eprint_id": 44042, "eprint_status": "archive", "datestamp": "2023-08-22 11:36:12", "lastmod": "2023-10-26 00:08:21", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Marotta-C-B", "name": { "family": "Marotta", "given": "Christopher B." } }, { "id": "Dilworth-C-N", "name": { "family": "Dilworth", "given": "Crystal N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Probing the non-canonical interface for agonist interaction with an \u03b15 containing nicotinic acetylcholine receptor", "ispublished": "pub", "full_text_status": "public", "keywords": "Addiction; Biophysics; Cholinergic receptor; Electrophysiology; Ion channels; Nicotinic acetylcholine receptors", "note": "\u00a9 2013 Elsevier Ltd.\n\nReceived 19 July 2013; Received in revised form 25 September 2013; Accepted 30 September 2013.\n\nWe thank Bruce N. Cohen for help with data interpretation.\nThis work was supported by grants from the NIH (NS034407, DA017279, DA280382), NIH/NRSA (GM07616) and the California Tobacco-Related Disease Research Program from the University of California (19XT-0102).\n\nAccepted Version - nihms-533201.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) containing the \u03b15 subunit are of interest because genomewide\nassociation studies and candidate gene studies have identified polymorphisms in the \u03b15 gene\nthat are linked to an increased risk for nicotine dependence, lung cancer, and/or alcohol addiction. To\nprobe the functional impact of an \u03b15 subunit on nAChRs, a method to prepare a homogeneous population\nof \u03b15-containing receptors must be developed. Here we use a gain of function (9') mutation to isolate\npopulations of \u03b15-containing nAChRs for characterization by electrophysiology. We find that the \u03b15\nsubunit modulates nAChR rectification when co-assembled with \u03b14 and \u03b22 subunits. We also probe the\n\u03b15-\u03b14 interface for possible ligand-binding interactions. We find that mutations expected to ablate an\nagonist-binding site involving the \u03b15 subunit have no impact on receptor function. The most straightforward\ninterpretation of this observation is that agonists do not bind at the \u03b15-\u03b14 interface, in contrast\nto what has recently been demonstrated for the \u03b14-\u03b14 interface in related receptors. In addition, our\nmutational results suggest that the \u03b15 subunit does not replace the \u03b14 or \u03b22 subunits and is relegated to\noccupying only the auxiliary position of the pentameric receptor.", "date": "2014-02", "date_type": "published", "publication": "Neuropharmacology", "volume": "77", "publisher": "Elsevier", "pagerange": "342-349", "id_number": "CaltechAUTHORS:20140227-142503676", "issn": "0028-3908", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140227-142503676", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "DA280382" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM07616" }, { "agency": "University of California California Tobacco-Related Disease Research Program", "grant_number": "19XT-0102" } ] }, "doi": "10.1016/j.neuropharm.2013.09.028", "pmcid": "PMC3934363", "primary_object": { "basename": "nihms-533201.pdf", "url": "https://authors.library.caltech.edu/records/501sm-h7s17/files/nihms-533201.pdf" }, "resource_type": "article", "pub_year": "2014", "author_list": "Marotta, Christopher B.; Dilworth, Crystal N.; et el." }, { "id": "https://authors.library.caltech.edu/records/ry04d-rnn52", "eprint_id": 47127, "eprint_status": "archive", "datestamp": "2023-08-19 23:15:50", "lastmod": "2023-10-26 20:20:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Marotta-C-B", "name": { "family": "Marotta", "given": "Christopher" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Identifying Motifs Essential for Selective Activation of Nicotinic Acetylcholine Receptors", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2014 Biophysical Society. Published by Elsevier Inc.", "abstract": "Neuronal Nicotinic acetylcholine receptors (nAChRs) are found throughout the brain and have vital roles in memory and learning. In addition, these ligand gated ion channel are major targets of drug research for neurological disorders. Here, the two stoichiometries of \u03b14\u03b22 nAChRs expressed in Xenopus oocytes were probed using two agonists that display stoichiometry selectivity: Sazetidine A and NS9283. The results from these studies led to the following conclusions that may be generalized to the nAChR family: 1) an agonist must be bound to each \u03b1 subunit in the receptor in order to fully activate the receptor and 2) three key residues located on the complementary side of the subunit dictate agonist selectivity. Through better understanding of agonist binding, more potent and selective responses with minimized off target responses can be obtained.", "date": "2014-01-28", "date_type": "published", "publication": "Biophysical Journal", "volume": "106", "number": "2", "publisher": "Biophysical Society", "pagerange": "341A", "id_number": "CaltechAUTHORS:20140710-092155936", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140710-092155936", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/j.bpj.2013.11.1951", "resource_type": "article", "pub_year": "2014", "author_list": "Marotta, Christopher; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/d1w00-hre55", "eprint_id": 47133, "eprint_status": "archive", "datestamp": "2023-08-19 23:16:28", "lastmod": "2023-10-26 20:20:35", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Post-M-R", "name": { "family": "Post", "given": "Michael R." }, "orcid": "0000-0002-3214-7619" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Structure Function Studies at Two Different Nicotinic Acetylcholine Receptor Subtypes", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2014 Biophysical Society. Published by Elsevier Inc.", "abstract": "The nicotinic acetylcholine receptor (nAChR) is involved with regulating the release of neurotransmitter in the synapse. This research focuses on two subtypes of neuronal nAChR, the \u03b16\u03b22 and \u03b14\u03b22 receptors. Subtypes containing the \u03b16 and \u03b22 subunits are found mainly in dopaminergic neurons. The acetylcholine binding site in these subtypes, which sits at the interface of the two subunits, is an important therapeutic target. Expression and activation of a pure population of \u03b16\u03b22 receptors in X. laevis oocytes has previously never been reported. This study used two mutations in the \u03b22 M3-M4 loop and a reporter mutation in the M2 helix of each subunit to successfully express \u03b16\u03b22 receptors. Expression has been optimized, and a method to control subunit stoichiometry has been developed. We then used unnatural amino acid mutagenesis to develop a binding model for acetylcholine and nicotine at the \u03b16-\u03b22 subunit interface.\n\nThe \u03b14\u03b22 subtype is the most prevalent nAChR subtype found in the brain and is implicated in many neurological disorders. Previous work has established a hydrogen bond interaction between the pyrrolidine nitrogen of nicotine and the carbonyl backbone adjacent to Trp149 in the \u03b14 subunit. N'-methylnicotinium, which has a quaternary amine at the pyrrolidine nitrogen and cannot donate a hydrogen bond, was synthesized in order to probe this hydrogen bond donating interaction. Through a double-mutant cycle analysis utilizing an \u03b1-hydroxy acid mutation to perturb the carbonyl backbone hydrogen bond acceptor, we have determined that the hydrogen bonding interaction has a coupling energy of \u223c2 kcal/mol.", "date": "2014-01-28", "date_type": "published", "publication": "Biophysical Journal", "volume": "106", "number": "2", "publisher": "Biophysical Society", "pagerange": "341A", "id_number": "CaltechAUTHORS:20140710-095141475", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140710-095141475", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/j.bpj.2013.11.1950", "resource_type": "article", "pub_year": "2014", "author_list": "Post, Michael R.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/p8n88-7dv20", "eprint_id": 53340, "eprint_status": "archive", "datestamp": "2023-08-19 22:44:44", "lastmod": "2023-10-19 14:47:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry" }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Drug Actions on Nicotinic Receptors. Chronic vs Acute; Outside-in vs Inside-out", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2014 American College of Neuropsychopharmacology.", "abstract": "The superfamily of nicotinic acetylcholine\nreceptors (nAChRs) comprises various homopentameric and heteropentameric ligand-gated channels (such as alpha7\nand alpha4beta2*, respectively). Continuing studies show\nhow the acute effects of exposure to nicotine (ms to min)\narise from an ''outside-in'' event: activation and desensitization\nof sodium, potassium, and calcium selective ion channels\nin the various nAChRs. The ''outside-in'' events of nicotine\nresemble the actions of acetylcholine itself. These events are\nbecoming known on the neuroanatomical scale of micrometers,\nthe time scale of ms, the structural distance scale ofA \u00b0 ,\nand the genomic resolution of single bp.\nMethods: Evolving methods are also revealing new features of\nnicotinic drug action during **chronic** exposure (hr to wk).\nThe new methods include fluorescently labeled nAChRs,\nexpressed both in knock-in mice and in cultured model\nsystems; proteomics of molecules that interact with nAChRs;\nand RNA-Seq of genes activated by chronic nicotine.\nResults: An ''inside-out'' mechanism is becoming a likely\nmechanism to explain some chronic actions of nicotine and\nother nicotinic drugs. The \"inside-out\" pathway occurs at\nmuch lower nicotine concentrations than channel activation.\nThe ''inside-out'' pathway begins with pharmacological\nchaperoning: nicotine and related drugs permeate into\nthe cytoplasm, then into the endoplasmic reticulum. There,\nthe drugs interact with nAChRs and stabilize some nascent\nnAChRs. The best-known result of \"inside-out\" action is the\nclassical posttranslational ''upregulation'' of some nAChRs.\nIn addition, the ''inside-out'' pathway also leads, via\npharmacological chaperoning, to other events including\ndecreased endoplasmic reticulum stress, decreased unfolded\nprotein response, and possibly perturbed nAChR interactions\nwith other molecules.\nConclusions: The ''inside-events'' are not straightforward\ncontinuations of transduction pathways activated during\n''outside-in'' activation and desensitization, but result when\nprolonged (hr to wk) nicotine binding to intracellular\nnAChRs activates an entirely different set of pathways. The\n''inside-out'' pathway is accreting many biophysical,\nbiochemical, thermodynamic, cell biological, neuroanatomical,\nand electrophysiological details. Yet, ''inside-out''\nmechanisms have arisen, in part, from re-examining\nclassical pharmacokinetic and pharmacodynamic facts:\nnicotine can interact with intracellular molecules many\norders of magnitude more strongly than can the endogenous\nneurotransmitter, acetylcholine. There is no current\nevidence that the ''inside-out'' events have counterparts\nin the normal cell or organismic biology of nAChRs. The\npresent challenge is pursue the hypothesis that the ''insideout''\nevents have the power and selectivity to underlie\ntwo aspects of chronic exposure to nicotine: nicotine\ndependence and apparent neuroprotection.\nDisclosure: Nothing to Disclose.", "date": "2014", "date_type": "published", "publication": "Neuropsychopharmacology", "volume": "39", "number": "S1", "publisher": "Nature Publishing Group", "pagerange": "581-582", "id_number": "CaltechAUTHORS:20150108-114406249", "issn": "0893-133X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150108-114406249", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "resource_type": "article", "pub_year": "2014", "author_list": "Lester, Henry" }, { "id": "https://authors.library.caltech.edu/records/xkpgn-3ab31", "eprint_id": 43497, "eprint_status": "archive", "datestamp": "2023-08-19 22:40:35", "lastmod": "2023-10-25 23:36:04", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Henderson-B-J", "name": { "family": "Henderson", "given": "Brandon J." }, "orcid": "0000-0003-0381-028X" }, { "id": "Srinivasan-Rahul", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Nichols-W-A", "name": { "family": "Nichols", "given": "Weston A." } }, { "id": "Dilworth-C-N", "name": { "family": "Dilworth", "given": "Crystal N." } }, { "id": "Gutierrez-D-F", "name": { "family": "Gutierrez", "given": "Diana F." } }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "McKinney-Sheri", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Richards-C-I", "name": { "family": "Richards", "given": "Christopher I." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Nicotine exploits a COPI-mediated process for chaperone-mediated up-regulation of its receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2014 Henderson et al. This article is distributed under the terms of an Attribution\u2013Noncommercial\u2013Share Alike\u2013No Mirror Sites license for the first six months after the publication\ndate (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).\n\nSubmitted: 16 September 2013; accepted: 6 December 2013.\n\nPublished December 30, 2013.\nWe thank Jennifer Lippincott-Schwartz for kindly providing \u025bCOPGFP subunits for our studies, Rell Parker for comments, and the Barbara Wold laboratory for providing cryosectioning tools. This work was supported by National Institutes of Health (grants AG033954, DA017279, DA019375, DA030396, NS034407, and DA033721) and by the California Tobacco-Related Disease Research Program (grant 17RT0127). Louis and Janet Fletcher provided partial funding for the TIRF microscope. The authors have no conflicting financial interests.\nEdward N. Pugh Jr. served as editor.\n\nPublished - J_Gen_Physiol-2014-Henderson-51-66.pdf
Supplemental Material - JGP_201311102_sm.pdf
", "abstract": "Chronic exposure to nicotine up-regulates high sensitivity nicotinic acetylcholine receptors (nAChRs) in the brain. This up-regulation partially underlies addiction and may also contribute to protection against Parkinson's disease. nAChRs containing the \u03b16 subunit (\u03b16* nAChRs) are expressed in neurons in several brain regions, but comparatively little is known about the effect of chronic nicotine on these nAChRs. We report here that nicotine up-regulates \u03b16* nAChRs in several mouse brain regions (substantia nigra pars compacta, ventral tegmental area, medial habenula, and superior colliculus) and in neuroblastoma 2a cells. We present evidence that a coat protein complex I (COPI)-mediated process mediates this up-regulation of \u03b16* or \u03b14* nAChRs but does not participate in basal trafficking. We show that \u03b16\u03b22\u03b23 nAChR up-regulation is prevented by mutating a putative COPI-binding motif in the \u03b23 subunit or by inhibiting COPI. Similarly, a COPI-dependent process is required for up-regulation of \u03b14\u03b22 nAChRs by chronic nicotine but not for basal trafficking. Mutation of the putative COPI-binding motif or inhibition of COPI also results in reduced normalized F\u00f6rster resonance energy transfer between \u03b16\u03b22\u03b23 nAChRs and \u03b5COP subunits. The discovery that nicotine exploits a COPI-dependent process to chaperone high sensitivity nAChRs is novel and suggests that this may be a common mechanism in the up-regulation of nAChRs in response to chronic nicotine.", "date": "2013-12-30", "date_type": "published", "publication": "Journal of General Physiology", "volume": "143", "number": "1", "publisher": "Rockefeller University Press", "pagerange": "51-66", "id_number": "CaltechAUTHORS:20140123-151832351", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20140123-151832351", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "DA019375" }, { "agency": "NIH", "grant_number": "DA030396" }, { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "NIH", "grant_number": "DA033721" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "17RT0127" } ] }, "doi": "10.1085/jgp.201311102", "pmcid": "PMC3874574", "primary_object": { "basename": "JGP_201311102_sm.pdf", "url": "https://authors.library.caltech.edu/records/xkpgn-3ab31/files/JGP_201311102_sm.pdf" }, "related_objects": [ { "basename": "J_Gen_Physiol-2014-Henderson-51-66.pdf", "url": "https://authors.library.caltech.edu/records/xkpgn-3ab31/files/J_Gen_Physiol-2014-Henderson-51-66.pdf" } ], "resource_type": "article", "pub_year": "2013", "author_list": "Henderson, Brandon J.; Srinivasan, Rahul; et el." }, { "id": "https://authors.library.caltech.edu/records/1zh8z-ewp74", "eprint_id": 39932, "eprint_status": "archive", "datestamp": "2023-08-19 21:54:45", "lastmod": "2023-10-24 17:19:05", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Henley-B-M", "name": { "family": "Henley", "given": "Beverley M." } }, { "id": "Williams-B-A", "name": { "family": "Williams", "given": "Brian A." } }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "Wold-B-J", "name": { "family": "Wold", "given": "Barbara J." }, "orcid": "0000-0003-3235-8130" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Transcriptional regulation by nicotine in dopaminergic neurons", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Nicotinic acetylcholine receptor, RNA-Seq, Substantia nigra pars compacta, Dopamine-related genes, Ubiquitin-proteasome pathway", "note": "\u00a9 2013 Elsevier B.V. \n\nReceived 8 May 2013. Accepted 26 July 2013. Available online 9 August 2013. \n\nWe thank Igor Antoshechkin for library sequencing, sequencing facility management and for computational training, Sheri McKinney for excellent cell cultures and for conducting nicotine administration, and Purnima Deshpande for animal breeding and laboratory management. We thank Prof. John Allman for use of the Zeiss laser capture microscope, Georgi Marinov for some analysis and helpful discussions and Charlotte Yang for initial DAVID analysis. This work was supported by the NIH (DA017279, AG033954), by the California Tobacco-Related Disease Research Program, by the Caltech Innovation Initiative, and by the Millard and Muriel Jacobs Genetics and Genomics Laboratory at California Institute of Technology. We thank the reviewers for helpful and insightful comments.", "abstract": "Dopaminergic neurons in the substantia nigra pars compacta (SNc) degenerate in Parkinson's disease. These neurons robustly express several nicotinic acetylcholine receptor (nAChR) subtypes. Smoking appears to be neuroprotective for Parkinson's disease but the mechanism is unknown. To determine whether chronic nicotine-induced changes in gene expression contribute to the neuroprotective effects of smoking, we develop methods to measure the effect of prolonged nicotine exposure on the SNc neuronal transcriptome in an unbiased manner. Twenty neurons were collected using laser-capture microscopy and transcriptional changes were assessed using RNA deep sequencing. These results are the first whole-transcriptome analyses of chronic nicotinic treatment in SNc neurons. Overall, 129 genes were significantly regulated: 67 upregulated, 62 downregulated. Nicotine-induced relief of endoplasmic reticulum (ER) stress has been postulated as a potential mechanism for the neuroprotective effects of smoking. Chronic nicotine did not significantly affect the expression of ER stress-related genes, nor of dopamine-related or nAChR genes, but it did modulate expression of 129 genes that could be relevant to the neuroprotective effects of smoking, including genes involved in (1) the ubiquitin-proteasome pathway, (2) cell cycle regulation, (3) chromatin modification, and (4) DNA binding and RNA regulation. We also report preliminary transcriptome data for single-cell dopaminergic and GABAergic neurons isolated from midbrain cultures. These novel techniques will facilitate advances in understanding the mechanisms taking place at the cellular level and may have applications elsewhere in the fields of neuroscience and molecular biology. The results give an emerging picture of the role of nicotine on the SNc and on dopaminergic neurons.", "date": "2013-10-15", "date_type": "published", "publication": "Biochemical Pharmacology", "volume": "86", "number": "8", "publisher": "Elsevier", "pagerange": "1074-1083", "id_number": "CaltechAUTHORS:20130815-073947290", "issn": "0006-2952", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130815-073947290", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Caltech Innovation Initiative (CI2)" }, { "agency": "NIH", "grant_number": "MH086383" }, { "agency": "Millard and Muriel Jacobs Genetics and Genomics Laboratory" } ] }, "local_group": { "items": [ { "id": "Millard-and-Muriel-Jacobs-Genetics-and-Genomics-Laboratory" } ] }, "doi": "10.1016/j.bcp.2013.07.031", "pmcid": "PMC3969234", "resource_type": "article", "pub_year": "2013", "author_list": "Henley, Beverley M.; Williams, Brian A.; et el." }, { "id": "https://authors.library.caltech.edu/records/ke5m8-6sn80", "eprint_id": 41299, "eprint_status": "archive", "datestamp": "2023-08-19 21:07:19", "lastmod": "2023-10-24 23:37:29", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Miles-T-F", "name": { "family": "Miles", "given": "Timothy F." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "The 5-HT_3AB Receptor Shows an A_3B_2 Stoichiometry at the Plasma Membrane", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2013 Biophysical Society. Submitted May 16, 2013, and accepted for publication July 11, 2013. Editor: Cynthia Czajkowski. The authors thank Rahul Srinivasan, Kristina N. Daeffler, and Ethan B. Van Arnam for technical assistance and helpful discussion. This work was supported by grants (No. NS034407, No. DA017279, and No. AG033954) from the National Institutes of Health, Bethesda, MD.\n\nPublished - 1-s2.0-S0006349513007996-main.pdf
Supplemental Material - mmc1.pdf
", "abstract": "The 5-HT_3AB receptor is the best-characterized heteropentameric 5-HT_3 receptor. Under conditions of heterologous expression, the 5-HT_3AB receptor shows a single functionally resolvable population, suggesting the presence of a unique subunit stoichiometry; however, conflicting previous reports have suggested two different possible stoichiometries. Here we isolate plasma membrane sheets containing assembled receptors from individual HEK293T cells. We then determine the stoichiometry of 5-HT_3AB receptors on the plasma membrane by fluorescence methods, employing meCFP- and meYFP-labeled A and B subunits. Over a wide range of cDNA transfection ratios, fluorescence intensity ratios are closest to values that correspond to a subunit ratio of A_3B_2. F\u00f6rster resonance energy transfer (family FRET) efficiencies provide minor corrections (3\u20136%) to the subunit ratios and provide independent support for a predominantly A_3B_2 stoichiometry on the plasma membrane sheets. Twin FRET efficiencies support these data, also suggesting that the two B subunits are nonadjacent in most of the heteropentamers. The high-frequency variant HTR3B p.Y129S (c.386A>C, rs11767445), linked to psychiatric disease, also forms A_3B_2 receptors on the plasma membrane. The 5-HT_3B Y129S, subunit incorporates in a slightly (11\u201314%) more efficient manner than the common variant. In general, most of the subunits reside within the cell. In contrast to the findings for the plasma membrane, the relative abundances and FRET characteristics of intracellular subunits depend strongly on the transfection ratio. The straightforward and unambiguous combination of plasma membrane-sheet isolation, fluorescence intensity ratios, and FRET is a generally promising procedure for determining stoichiometry of proteins on the plasma membrane.", "date": "2013-08-20", "date_type": "published", "publication": "Biophysical Journal", "volume": "105", "number": "4", "publisher": "Biophysical Society", "pagerange": "887-898", "id_number": "CaltechAUTHORS:20130912-145808068", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130912-145808068", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "AG033954" } ] }, "doi": "10.1016/j.bpj.2013.07.015", "pmcid": "PMC3752109", "primary_object": { "basename": "1-s2.0-S0006349513007996-main.pdf", "url": "https://authors.library.caltech.edu/records/ke5m8-6sn80/files/1-s2.0-S0006349513007996-main.pdf" }, "related_objects": [ { "basename": "mmc1.pdf", "url": "https://authors.library.caltech.edu/records/ke5m8-6sn80/files/mmc1.pdf" } ], "resource_type": "article", "pub_year": "2013", "author_list": "Miles, Timothy F.; Dougherty, Dennis A.; et el." }, { "id": "https://authors.library.caltech.edu/records/z5smk-dpq65", "eprint_id": 39824, "eprint_status": "archive", "datestamp": "2023-08-19 20:53:42", "lastmod": "2023-10-24 17:13:39", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Van-Arman-E-B", "name": { "family": "Van Arnam", "given": "Ethan B." } }, { "id": "Blythe-E-E", "name": { "family": "Blythe", "given": "Emily E." }, "orcid": "0000-0001-6363-2644" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "An Unusual Pattern of Ligand-Receptor Interactions for the \u03b17 Nicotinic Acetylcholine Receptor, with Implications for the Binding of Varenicline", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2013 The American Society for Pharmacology and Experimental Therapeutics. \n\nReceived February 20, 2013; accepted May 16, 2013. \n\nThis work was supported by the National Institutes of Health National Institute of Neurological Disorders and Stroke [Grant NS34407]; and California Tobacco-Related Disease Research Program of the University of California, Award 19XT-0102. E.E.B. was an Amgen Scholar. The authors thank Pfizer for a generous gift of varenicline.\nParticipated in research design: Van Arnam, Blythe, Lester, Dougherty. Conducted experiments: Van Arnam, Blythe. Performed data analysis: Van Arnam, Blythe, Lester, Dougherty. Wrote or contributed to the writing of the manuscript: Van Arnam, Dougherty.\n\nSupplemental Material - Van_Arnam_et_al_Supplemental_Rev.pdf
", "abstract": "The \u03b17 nicotinic acetylcholine receptor shows broad pharmacology, complicating the development of subtype-specific nicotinic receptor agonists. Here we use unnatural amino acid mutagenesis to characterize binding to \u03b17 by the smoking cessation drug varenicline (Chantix; Pfizer, Groton, CT), an \u03b14\u03b22-targeted agonist that shows full efficacy and modest potency at the \u03b17 receptor. We find that unlike binding to its target receptor, varenicline does not form a cation-\u03c0 interaction with TrpB, further supporting a unique binding mode for the cationic amine of nicotinic agonists at the \u03b17 receptor. We also evaluate binding to the complementary face of the receptor's binding site by varenicline, the endogenous agonist acetylcholine, and the potent nicotine analog epibatidine. Interestingly, we find no evidence for functionally important interactions involving backbone NH and CO groups thought to bind the canonical agonist hydrogen bond acceptor of the nicotinic pharmacophore, perhaps reflecting a lesser importance of this pharmacophore element for \u03b17 binding. We also show that the Trp55 and Leu119 side chains of the binding site's complementary face are important for the binding of the larger agonists epibatidine and varenicline, but dispensable for binding of the smaller, endogenous agonist acetylcholine.", "date": "2013-08", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "84", "number": "2", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "201-207", "id_number": "CaltechAUTHORS:20130808-161341375", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130808-161341375", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19XT-0102" }, { "agency": "Amgen" }, { "agency": "National Institute of Neurological Disorders and Stroke (NINDS)" } ] }, "doi": "10.1124/mol.113.085795", "pmcid": "PMC3716316", "primary_object": { "basename": "Van_Arnam_et_al_Supplemental_Rev.pdf", "url": "https://authors.library.caltech.edu/records/z5smk-dpq65/files/Van_Arnam_et_al_Supplemental_Rev.pdf" }, "resource_type": "article", "pub_year": "2013", "author_list": "Van Arnam, Ethan B.; Blythe, Emily E.; et el." }, { "id": "https://authors.library.caltech.edu/records/rrxy5-gke71", "eprint_id": 41160, "eprint_status": "archive", "datestamp": "2023-08-19 20:48:19", "lastmod": "2023-10-24 23:30:02", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Frazier-S-J", "name": { "family": "Frazier", "given": "Shawnalea J." } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "An Engineered Glutamate-gated Chloride (GluCl) Channel for Sensitive, Consistent Neuronal Silencing by Ivermectin", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2013 by The American Society for Biochemistry and Molecular Biology, Inc.\n\nReceived for publication, September 28, 2012, and in revised form, May 27, 2013 Published, JBC Papers in Press, May 29, 2013.\n\nThis work was supported, in whole or in part, by National Institutes of Health\nGrants NS034407, EY018502, and MH088550. This work was also supported\nby the McKnight Foundation.\n\nWe thank Sheri McKinney for providing hippocampal\nneuron cultures. We also thank D. A. Dougherty and D.\nAnderson for comments.\n\nPublished - J._Biol._Chem.-2013-Frazier-21029-42.pdf
", "abstract": "A modified invertebrate glutamate-gated Cl\u2212 channel (GluCl \u03b1\u03b2) was previously employed to allow pharmacologically induced silencing of electrical activity in CNS neurons upon exposure to the anthelmintic drug ivermectin (IVM). Usefulness of the previous receptor was limited by 1) the high concentration of IVM necessary to elicit a consistent silencing phenotype, raising concern about potential side effects, and 2) the variable extent of neuronal spike suppression, due to variations in the co-expression levels of the fluorescent protein-tagged \u03b1 and \u03b2 subunits. To address these issues, mutant receptors generated via rational protein engineering strategies were examined for improvement. Introduction of a gain-of-function mutation (L9\u2032F) in the second transmembrane domain of the \u03b1 subunit appears to facilitate \u03b2 subunit incorporation and substantially increase heteromeric GluCl \u03b1\u03b2 sensitivity to IVM. Removal of an arginine-based endoplasmic reticulum retention motif (RSR mutated to AAA) from the intracellular loop of the \u03b2 subunit further promotes heteromeric expression at the plasma membrane possibly by preventing endoplasmic reticulum-associated degradation of the \u03b2 subunit rather than simply reducing endoplasmic reticulum retention. A monomeric XFP (mXFP) mutation that prevents fluorescent protein dimerization complements the mutant channel effects. Expression of the newly engineered GluCl opt \u03b1-mXFP L9\u2032F + opt \u03b2-mXFP Y182F RSR_AAA receptor in dissociated neuronal cultures markedly increases conductance and reduces variability in spike suppression at 1 nm IVM. This receptor, named \"GluClv2.0,\" is an improved tool for IVM-induced silencing.", "date": "2013-07-19", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "288", "number": "29", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "21029-21042", "id_number": "CaltechAUTHORS:20130909-090111946", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130909-090111946", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS034407" }, { "agency": "NIH", "grant_number": "EY018502" }, { "agency": "NIH", "grant_number": "MH088550" }, { "agency": "McKnight Foundation" } ] }, "doi": "10.1074/jbc.M112.423921", "pmcid": "PMC3774370", "primary_object": { "basename": "J._Biol._Chem.-2013-Frazier-21029-42.pdf", "url": "https://authors.library.caltech.edu/records/rrxy5-gke71/files/J._Biol._Chem.-2013-Frazier-21029-42.pdf" }, "resource_type": "article", "pub_year": "2013", "author_list": "Frazier, Shawnalea J.; Cohen, Bruce N.; et el." }, { "id": "https://authors.library.caltech.edu/records/823wk-b3380", "eprint_id": 41170, "eprint_status": "archive", "datestamp": "2023-08-19 20:46:26", "lastmod": "2023-10-24 23:30:25", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Chatterjee-S", "name": { "family": "Chatterjee", "given": "Susmita" }, "orcid": "0000-0002-2878-1502" }, { "id": "Santos-N", "name": { "family": "Santos", "given": "Nathan" } }, { "id": "Holgate-J", "name": { "family": "Holgate", "given": "Joan" } }, { "id": "Haass-Koffler-C-L", "name": { "family": "Haass-Koffler", "given": "Carolina L." } }, { "id": "Hopf-F-W", "name": { "family": "Hopf", "given": "F. Woodward" } }, { "id": "Kharazia-V", "name": { "family": "Kharazia", "given": "Viktor" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry" }, "orcid": "0000-0002-5470-5255" }, { "id": "Bonci-A", "name": { "family": "Bonci", "given": "Antonello" } }, { "id": "Bartlett-S-E", "name": { "family": "Bartlett", "given": "Selena E." } } ] }, "title": "The \u03b15 Subunit Regulates the Expression and Function of \u03b14*-Containing Neuronal Nicotinic Acetylcholine Receptors in the Ventral-Tegmental Area", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2013 Chatterjee et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits\nunrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.\n\nReceived February 24, 2012; Accepted June 2, 2013; Published July 15, 2013.\n\nEditor: Zhong-Ping Feng, University of Toronto, Canada.\n\nFunding: This work was supported by funding from the National Institutes of Health (NIH) 1RC2AA019429-01 (to SEB), NIH 1R01AA017924-01 (to SEB),\nNIH 1R01DA17279 (to HL), and the State of California for Medical Research on Alcohol and Substance Abuse through the University of California San\nFrancisco (to SEB), and the California Tobacco-Related Disease Research Program (to HL). The funders had no role in study design, data collection and\nanalysis, decision to publish, or preparation of the manuscript.\n\nWe thank Stacy Taylor for excellent technical assistance in\nweaning and maintaining the breeding colony. We also thank\nSan Francisco Nikon Imaging Center, at the University of\nCalifornia San Francisco for assistance with imaging.\n\nAuthor Contributions:\nAnalyzed the data: JH CLHK SC. Wrote the manuscript: SC.\nConceived the project: SEB AB. Design of the study and\ndevelopment of the manuscript: SC SEB. Performed\nelectrophysiology experiments and analyzed data: SC.\nPerformed biochemistry experiments: JH CLHK VK. Provided\ncritical supplemental content and revisions: SC NS FWH VK\nSEB. Provided the \u03b14-YFP mice: HL. Provided significant\ncritiques to the manuscript: AB SEB FWH HL. Reviewed\ncontents of the study and have approved final version for\npublications: SEB SC NS JH CLHK FWH VK HL AB.\n\nPublished - journal.pone.0068300.pdf
", "abstract": "Human genetic association studies have shown gene variants in the \u03b15 subunit of the neuronal nicotinic receptor (nAChR) influence both ethanol and nicotine dependence. The \u03b15 subunit is an accessory subunit that facilitates \u03b14* nAChRs assembly in vitro. However, it is unknown whether this occurs in the brain, as there are few research tools to adequately address this question. As the \u03b14*-containing nAChRs are highly expressed in the ventral tegmental area (VTA) we assessed the molecular, functional and pharmacological roles of \u03b15 in \u03b14*-containing nAChRs in the VTA. We utilized transgenic mice \u03b15+/+(\u03b14YFP) and \u03b15-/-(\u03b14YFP) that allow the direct visualization and measurement of \u03b14-YFP expression and the effect of the presence (\u03b15+/+) and absence of \u03b15 (-/-) subunit, as the antibodies for detecting the \u03b14* subunits of the nAChR are not specific. We performed voltage clamp electrophysiological experiments to study baseline nicotinic currents in VTA dopaminergic neurons. We show that in the presence of the \u03b15 subunit, the overall expression of \u03b14 subunit is increased significantly by 60% in the VTA. Furthermore, the \u03b15 subunit strengthens baseline nAChR currents, suggesting the increased expression of \u03b14* nAChRs to be likely on the cell surface. While the presence of the \u03b15 subunit blunts the desensitization of nAChRs following nicotine exposure, it does not alter the amount of ethanol potentiation of VTA dopaminergic neurons. Our data demonstrates a major regulatory role for the \u03b15 subunit in both the maintenance of \u03b14*-containing nAChRs expression and in modulating nicotinic currents in VTA dopaminergic neurons. Additionally, the \u03b15\u03b14* nAChR in VTA dopaminergic neurons regulates the effect of nicotine but not ethanol on currents. Together, the data suggest that the \u03b15 subunit is critical for controlling the expression and functional role of a population of \u03b14*-containing nAChRs in the VTA.", "date": "2013-07-15", "date_type": "published", "publication": "PLoS ONE", "volume": "8", "number": "7", "publisher": "Public Library of Science", "pagerange": "Art. No. e68300", "id_number": "CaltechAUTHORS:20130909-105437251", "issn": "1932-6203", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130909-105437251", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "1RC2AA019429-01" }, { "agency": "NIH", "grant_number": "1R01AA017924-01" }, { "agency": "NIH", "grant_number": "1R01DA17279" }, { "agency": "University of California San Francisco State of California for Medical Research on Alcohol and Substance Abuse" }, { "agency": "California Tobacco-Related Disease Research Program" } ] }, "doi": "10.1371/journal.pone.0068300", "pmcid": "PMC3712017", "primary_object": { "basename": "journal.pone.0068300.pdf", "url": "https://authors.library.caltech.edu/records/823wk-b3380/files/journal.pone.0068300.pdf" }, "resource_type": "article", "pub_year": "2013", "author_list": "Chatterjee, Susmita; Santos, Nathan; et el." }, { "id": "https://authors.library.caltech.edu/records/p9qm0-jq795", "eprint_id": 37887, "eprint_status": "archive", "datestamp": "2023-08-19 19:01:19", "lastmod": "2023-10-23 19:05:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Blum-A-P", "name": { "family": "Blum", "given": "Angela P." } }, { "id": "Van-Arman-E-B", "name": { "family": "Van Arnam", "given": "Ethan B." } }, { "id": "German-L-A", "name": { "family": "German", "given": "Laurel A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Binding Interactions with the Complementary Subunit of Nicotinic Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2013 American Society for Biochemistry and Molecular Biology, Inc.\n\nReceived for publication, November 26, 2012, and in revised form, January 23, 2013. Published, JBC Papers in Press, January 24, 2013.\n\nThis work was supported, in whole or in part, by National Institutes of Health Grants NS34407 and NS11756. This work was also supported by Tobacco-Related Disease Research Program Award 19XT-0102 from the University of California.\nAcknowledgment\u2014We thank Pfizer for the generous gift of\nvarenicline.\n\nPublished - J._Biol._Chem.-2013-Blum-6991-7.pdf
", "abstract": "The agonist-binding site of nicotinic acetylcholine receptors (nAChRs) spans an interface between two subunits of the pentameric receptor. The principal component of this binding site is contributed by an \u03b1 subunit, and it binds the cationic moiety of the nicotinic pharmacophore. The other part of the pharmacophore, a hydrogen bond acceptor, has recently been shown to bind to the complementary non-\u03b1 subunit via the backbone NH of a conserved Leu. This interaction was predicted by studies of ACh-binding proteins and confirmed by functional studies of the neuronal (CNS) nAChR, \u03b14\u03b22. The ACh-binding protein structures further suggested that the hydrogen bond to the backbone NH is mediated by a water molecule and that a second hydrogen bonding interaction occurs between the water molecule and the backbone CO of a conserved Asn, also on the non-\u03b1 subunit. Here, we provide new insights into the nature of the interactions between the hydrogen bond acceptor of nicotinic agonists and the complementary subunit backbone. We studied both the nAChR of the neuromuscular junction (muscle-type) and a neuronal subtype, (\u03b14)2(\u03b24)3. In the muscle-type receptor, both ACh and nicotine showed a strong interaction with the Leu NH, but the potent nicotine analog epibatidine did not. This interaction was much attenuated in the \u03b14\u03b24 receptor. Surprisingly, we found no evidence for a functionally significant interaction with the backbone carbonyl of the relevant Asn in either receptor with an array of agonists.", "date": "2013-03-08", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "288", "number": "10", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "6991-6997", "id_number": "CaltechAUTHORS:20130411-103637479", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130411-103637479", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19XT-0102" } ] }, "doi": "10.1074/jbc.M112.439968", "pmcid": "PMC3591609", "primary_object": { "basename": "J._Biol._Chem.-2013-Blum-6991-7.pdf", "url": "https://authors.library.caltech.edu/records/p9qm0-jq795/files/J._Biol._Chem.-2013-Blum-6991-7.pdf" }, "resource_type": "article", "pub_year": "2013", "author_list": "Blum, Angela P.; Van Arnam, Ethan B.; et el." }, { "id": "https://authors.library.caltech.edu/records/1kkdy-zxe50", "eprint_id": 37670, "eprint_status": "archive", "datestamp": "2023-08-22 08:33:23", "lastmod": "2023-10-23 17:55:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Limapichat-W", "name": { "family": "Limapichat", "given": "Walrati" } }, { "id": "Yu-Wesley-Y", "name": { "family": "Yu", "given": "Wesley Y." } }, { "id": "Branigan-E", "name": { "family": "Branigan", "given": "Emma" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Key Binding Interactions for Memantine in the NMDA Receptor", "ispublished": "pub", "full_text_status": "public", "keywords": "Memantine; open-channel blocker; amantadine; unnatural amino acid mutagenesis", "note": "\u00a9 2012 American Chemical Society. \n\nReceived: October 12, 2012; Accepted: November 29, 2012; Published: November 29, 2012. \n\nThis work was supported by the NIH (NS 34407 to D.A.D.). \n\nAuthor Contributions: Contributed to research design: W.L., E.B., W.Y., H.A.L., and D.A.D. conducted experiments: W.L., E.B., and W.Y. Performed data analysis: W.L., E.B., W.Y., H.A.L., and D.A.D. Writing of the manuscript: W.L., W.Y., H.A.L., and D.A.D. \n\nWe thank Dr. Kathryn McMenimen for helpful discussions.\n\nSupplemental Material - cn300180a_si_001.pdf
", "abstract": "Memantine (Namenda) is prescribed as a treatment for moderate to severe Alzheimer's Disease. Memantine functions by blocking the NMDA receptor, but the key binding interactions between drug and receptor are not fully elucidated. To determine key binding interactions of memantine, we made side-by-side comparisons of IC_(50) for memantine and amantadine, a structurally related drug, in the GluN1/GluN2B NMDA receptor. We identified hydrophobic binding pockets for the two methyl groups on memantine formed by the residues A645 and A644 on the third transmembrane helices of GluN1 and GluN2B, respectively. Moreover, we found that while adding two methyl groups to amantadine to produce memantine greatly improves affinity, adding a third methyl group to produce the symmetrical trimethylamantadine diminished affinity. Our results provide a better understanding of chemical-scale interactions between memantine and the NMDA channel, which will potentially benefit the development of new drugs for neurodegenerative diseases involving NMDA receptors.", "date": "2013-02", "date_type": "published", "publication": "ACS Chemical Neuroscience", "volume": "4", "number": "2", "publisher": "American Chemical Society", "pagerange": "255-260", "id_number": "CaltechAUTHORS:20130328-101734866", "issn": "1948-7193", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130328-101734866", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" } ] }, "doi": "10.1021/cn300180a", "pmcid": "PMC3751542", "primary_object": { "basename": "cn300180a_si_001.pdf", "url": "https://authors.library.caltech.edu/records/1kkdy-zxe50/files/cn300180a_si_001.pdf" }, "resource_type": "article", "pub_year": "2013", "author_list": "Limapichat, Walrati; Yu, Wesley Y.; et el." }, { "id": "https://authors.library.caltech.edu/records/6ejxh-cq403", "eprint_id": 37559, "eprint_status": "archive", "datestamp": "2023-08-22 08:06:25", "lastmod": "2023-10-23 17:45:22", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "O'Neill-H-C", "name": { "family": "O'Neill", "given": "Heidi C." } }, { "id": "Laverty-D-C", "name": { "family": "Laverty", "given": "Duncan C." } }, { "id": "Patzlaff-N-E", "name": { "family": "Patzlaff", "given": "Natalie E." } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Fonck-C", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "McKinney-S", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "McIntosh-J-M", "name": { "family": "McIntosh", "given": "J. Michael" } }, { "id": "Lindstrom-J-M", "name": { "family": "Lindstrom", "given": "Jon M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } } ] }, "title": "Mice expressing the ADNFLE valine 287 leucine mutation of the \u03922 nicotinic acetylcholine receptor subunit display increased sensitivity to acute nicotine administration and altered presynaptic nicotinic receptor function", "ispublished": "pub", "full_text_status": "public", "keywords": "Nicotinic acetylcholine receptor; Autosomal dominant nocturnal frontal lobe epilepsy; Nicotine; Autoradiography; Knock-in mouse; Synaptosomes", "note": "\u00a9 2012 Elsevier Inc. Received 25 July 2012. Received in revised form 26 September 2012. Accepted 24 October 2012.\nAvailable online 1 November 2012. The authors thank Jian Xuand Steve Heinemannatthe SalkInstitute, La Jolla, CA for initially supplying the \u03b22VL mutant mice. This work was supported by grants R01 DA003194, R01 DA012242, P30 DA015663 and U19 DA019375 to MJM; R01 GM 103801 and R01 GM 48677 to JMM; R01 NS 11323 to JML\n\nAccepted Version - nihms418937.pdf
", "abstract": "Several mutations in \u03b14 or \u03b22 nicotinic receptor subunits are linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE). One such missense mutation in the gene encoding the \u03b22 neuronal nicotinic acetylcholine receptor (nAChR) subunit (CHRNB2) is a valine-to-leucine substitution in the second transmembrane domain at position 287 (\u03b22VL). Previous studies indicated that the \u03b22VL mutation in mice alters circadian rhythm consistent with sleep alterations observed in ADNFLE patients (Xu et al., 2011). The current study investigates changes in nicotinic receptor function and expression that may explain the behavioral phenotype of \u03b22VL mice. No differences in \u03b22 mRNA expression were found between wild-type (WT) and heterozygous (HT) or homozygous mutant (MT) mice. However, antibody and ligand binding indicated that the mutation resulted in a reduction in receptor protein. Functional consequences of the \u03b22VL mutation were assessed biochemically using crude synaptosomes. A gene-dose dependent increase in sensitivity to activation by acetylcholine and decrease in maximal nAChR-mediated [^3H]-dopamine release and ^(86)Rb efflux were observed. Maximal nAChR-mediated [^3H]-GABA release in the cortex was also decreased in the MT, but maximal [^3H]-GABA release was retained in the hippocampus. Behaviorally both HT and MT mice demonstrated increased sensitivity to nicotine-induced hypolocomotion and hypothermia. Furthermore, WT mice display only a tonic\u2013clonic seizure (EEG recordable) 3 min after injection of a high dose of nicotine, while MT mice also display a dystonic arousal complex (non-EEG recordable) event 30 s after nicotine injection. Data indicate decreases in maximal response for certain measures are larger than expected given the decrease in receptor expression.", "date": "2013-01", "date_type": "published", "publication": "Pharmacology Biochemistry and Behavior", "volume": "103", "number": "3", "publisher": "Elsevier", "pagerange": "603-621", "id_number": "CaltechAUTHORS:20130319-103616459", "issn": "0091-3057", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130319-103616459", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "R01 DA003194" }, { "agency": "NIH", "grant_number": "R01 DA012242" }, { "agency": "NIH", "grant_number": "P30 DA015663" }, { "agency": "NIH", "grant_number": "U19 DA019375" }, { "agency": "NIH", "grant_number": "R01 GM 103801" }, { "agency": "NIH", "grant_number": "R01 GM 48677" }, { "agency": "NIH", "grant_number": "R01 NS 11323" } ] }, "doi": "10.1016/j.pbb.2012.10.014", "pmcid": "PMC3544981", "primary_object": { "basename": "nihms418937.pdf", "url": "https://authors.library.caltech.edu/records/6ejxh-cq403/files/nihms418937.pdf" }, "resource_type": "article", "pub_year": "2013", "author_list": "O'Neill, Heidi C.; Laverty, Duncan C.; et el." }, { "id": "https://authors.library.caltech.edu/records/jzmj6-vtw04", "eprint_id": 35998, "eprint_status": "archive", "datestamp": "2023-08-22 07:48:46", "lastmod": "2023-10-20 22:06:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Miwa-J-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } } ] }, "title": "Psychiatric Drugs Bind to Classical Targets Within Early Exocytotic Pathways: Therapeutic Effects", "ispublished": "pub", "full_text_status": "public", "keywords": "Antipsychotics; depression; gene activation; nicotine; schizophrenia; serotonin selective reuptake inhibitors", "note": "\u00a9 2012 Society of Biological Psychiatry. \n\nReceived Mar 21, 2012; revised Apr 23, 2012; accepted May 21, 2012. \n\nThis work was supported by the National Institutes of Health (MH-086383 and MH-088550). Our laboratory's work on nicotinic receptors has been supported by the National Institutes of Health (AG-33954, DA-17279, DA-19375), the Michael J. Fox Foundation, the Joint Center for Translational Medicine, the California Tobacco-Related Disease Research Program (17RT-0127, 18FT-0066, 19KT-0032), and Targacept, Inc. In the past, our laboratory's work on G protein-coupled receptors has been supported by GM-081662, on neurotransmitter transporters by DA-09121, and on intracellular messengers by GM-29836.\nHAL, JMM, and RS wrote the article. HAL developed the concepts.\nJMM contributed ideas on lynx. RS contributed scholarship on the unfolded\nprotein response.\nWe thank Hadassah Tamir for suggesting that axonal transport\nunderlies some aspects of psychiatric drug effects and for discussion,\nJohannes Schwarz for pointing out aspects of dyskinesia, and Luke\nWiseman for tutorials on the unfolded protein response.Wealso thank\nNathan Dascal, Dennis Dougherty, Hesso Farhan, Robert Farley, Robert\nFreedman, Ege Kavalali, Odile Kellermann, Jun Li,JohnLowe, Stefan\nMcDonough, J. Michael McIntosh, Randy Schekman, Darryle Schoepp,\nAndrew Steele, Teagan Wall, and the anonymous referees for comments.\nHAL thanks members of our research group for discussion and\nfor allowing the time to assemble this essay.\nHAL has submitted patent applications covering pharmacological\nchaperones in neurodegenerative and psychiatric disease. JMM is\nfounder and shareholder of Ophidion, Inc. She has applied for US patents\n10322359 and 20080221013 on the use of lynx for therapeutic\npurposes. RS reports no biomedical financial interests or potential conflicts of interest.\n\nAccepted Version - nihms-982735.pdf
Supplemental Material - mmc1.pdf
", "abstract": "The classical targets for antipsychotic and antidepressant drugs are G protein-coupled receptors and neurotransmitter transporters, respectively. Full therapeutic actions of these drugs require several weeks. We show how therapeutic effects may eventually accrue after existing therapeutic ligands bind to these classical targets, not on the plasma membrane but rather within endoplasmic reticulum (ER) and cis-Golgi. Consequences of such binding may include pharmacological chaperoning: the nascent drug targets are stabilized against degradation and can therefore exit the ER more readily. Another effect may be matchmaking: heterodimers and homodimers of the target form and can more readily exit the ER. Summarizing recent data for nicotinic receptors, we explain how such effects could lead to reduced ER stress and to a decreased unfolded protein response, including changes in gene activation and protein synthesis. In effects not directly related to cellular stress, escorting would allow increased ER exit and trafficking of known associated proteins, as well as other proteins such as growth factors and their receptors, producing both cell-autonomous and non-cell-autonomous effects. Axonal transport of relevant proteins may underlie the several weeks required for full therapy. In contrast, the antidepressant effects of ketamine and other N-methyl-D-aspartate receptor ligands, which occur within <2 hours, could arise from dendritically localized intracellular binding, followed by chaperoning, matchmaking, escorting, and reduced ER stress. Thus, the effects of intracellular binding extend beyond proteostasis of the targets themselves and involve pathways distinct from ion channel and G protein activation. We propose experimental tests and note pathophysiological correlates.", "date": "2012-12", "date_type": "published", "publication": "Biological Psychiatry", "volume": "72", "number": "11", "publisher": "Elsevier", "pagerange": "907-915", "id_number": "CaltechAUTHORS:20121217-074942263", "issn": "0006-3223", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20121217-074942263", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "MH-86383" }, { "agency": "NIH", "grant_number": "MH-088550" }, { "agency": "NIH", "grant_number": "AG-33954" }, { "agency": "NIH", "grant_number": "DA-17279" }, { "agency": "NIH", "grant_number": "DA-19375" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "Joint Center for Translational Medicine" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "17RT-0127" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "18FT-0066" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19KT-0032" }, { "agency": "Targacept, Inc." }, { "agency": "NIH", "grant_number": "GM-081662" }, { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "GM-29836" } ] }, "doi": "10.1016/j.biopsych.2012.05.020", "pmcid": "PMC6167061", "primary_object": { "basename": "mmc1.pdf", "url": "https://authors.library.caltech.edu/records/jzmj6-vtw04/files/mmc1.pdf" }, "related_objects": [ { "basename": "nihms-982735.pdf", "url": "https://authors.library.caltech.edu/records/jzmj6-vtw04/files/nihms-982735.pdf" } ], "resource_type": "article", "pub_year": "2012", "author_list": "Lester, Henry A.; Miwa, Julie M.; et el." }, { "id": "https://authors.library.caltech.edu/records/jwegq-dc938", "eprint_id": 35670, "eprint_status": "archive", "datestamp": "2023-08-19 13:11:51", "lastmod": "2023-10-20 20:09:08", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Miles-T-F", "name": { "family": "Miles", "given": "Timothy F." } }, { "id": "Bower-K-S", "name": { "family": "Bower", "given": "Kiowa S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "A Coupled Array of Noncovalent Interactions Impacts the Function of the 5\u2011HT_3A Serotonin Receptor in an Agonist-Specific Way", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Serotonin, mCPBG, Cys-loop, binding site, gating, unnatural amino acid mutagenesis", "note": "\u00a9 2012 American Chemical Society. Received: May 30, 2012.\nAccepted: July 20, 2012. Publication Date (Web): July 20, 2012. Author Contributions:\nT.F.M., K.S.B., H.A.L., and D.A.D. contributed to research\ndesign. T.F.M. and K.S.B. conducted experiments. T.F.M.,\nK.S.B., H.A.L., and D.A.D. performed data analysis. T.F.M.,\nK.S.B., H.A.L., and D.A.D. wrote the manuscript. The authors declare no competing financial interest. We thank Dr. Sarah Lummis (Cambridge) for helpful discussions and Noah Duffy for the preparation of 4,7-F2-Trp. This work was supported by the NIH (NS 34407 to D.A.D.).", "abstract": "The serotonin type 3A (5-HT_3A) receptor is a Cys-loop (pentameric) neurotransmitter-gated ion channel found in the central and peripheral nervous systems and implicated in numerous diseases. In previous studies with the endogenous agonist serotonin, we identified two interactions critical for receptor function: a cation\u2212\u03c0 interaction with W183 in loop B (TrpB) and a hydrogen bond to E129 in loop A. Here we employ mutant cycle analyses utilizing conventional and unnatural amino acid mutagenesis to demonstrate that a third residue, D124 of loop A, forms two functionally important hydrogen bonds to the backbone of loop B. We also show that these three interactions, the cation\u2212\u03c0 interaction, the backbone hydrogen bonds, and the E129 hydrogen bond, are tightly coupled to each other, suggesting they function as a single unit. We also identify key functional differences between serotonin and the competitive partial agonist m-chlorophenyl biguanide (mCPBG) at these residues. mCPBG displays no cation\u2212\u03c0 at TrpB and extreme sensitivity to the positioning of E129, on which it is reliant for initiation of channel gating.", "date": "2012-10-17", "date_type": "published", "publication": "ACS Chemical Neuroscience", "volume": "3", "number": "10", "publisher": "American Chemical Society", "pagerange": "753-760", "id_number": "CaltechAUTHORS:20121127-103848210", "issn": "1948-7193", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20121127-103848210", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" } ] }, "doi": "10.1021/cn3000586", "pmcid": "PMC3474281", "resource_type": "article", "pub_year": "2012", "author_list": "Miles, Timothy F.; Bower, Kiowa S.; et el." }, { "id": "https://authors.library.caltech.edu/records/nyy8d-kxg21", "eprint_id": 35999, "eprint_status": "archive", "datestamp": "2023-08-22 07:10:51", "lastmod": "2023-10-20 22:06:45", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Scott-K-M", "name": { "family": "Scott", "given": "Kimberly M." } }, { "id": "Du-Jiangang", "name": { "family": "Du", "given": "Jiangang" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Masmanidis-S-C", "name": { "family": "Masmanidis", "given": "Sotiris C." } } ] }, "title": "Variability of acute extracellular action potential measurements with multisite silicon probes", "ispublished": "pub", "full_text_status": "public", "keywords": "Silicon neural probes; Multielectrode arrays; MEMS", "note": "\u00a9 2012 Elsevier B.V. Received 9 April 2012. Received in revised form 31 July 2012. Accepted 4 August 2012. We thank T.J. Blanche for discussions on probe design and recording quality assessment, R.R. Harrison for developing the ASIC\nused to read out data from the probe, and M.L. Roukes for device fabrication support. Neural probe nanofabrication was carried out at the Kavli Nanoscience Institute at Caltech, and the Nanoelectronics Research Facility at UCLA. We acknowledge support from the Broad Foundations, the Della Martin Fund for Discoveries in Mental Illness, and the NIH (DA-17279 and AG-033954).\n\nAccepted Version - nihms-401238.pdf
", "abstract": "Device miniaturization technologies have led to significant advances in sensors for extracellular measurements of electrical activity in the brain. Multisite, silicon-based probes containing implantable electrode arrays afford greater coverage of neuronal activity than single electrodes and therefore potentially offer a more complete view of how neuronal ensembles encode information. However, scaling up the number of sites is not sufficient to ensure capture of multiple neurons, as action potential signals from extracellular electrodes may vary due to numerous factors. In order to understand the large-scale recording capabilities and potential limitations of multisite probes, it is important to quantify this variability, and to determine whether certain key device parameters influence the recordings. Here we investigate the effect of four parameters, namely, electrode surface, width of the structural support shafts, shaft number, and position of the recording site relative to the shaft tip. This study employs acutely implanted silicon probes containing up to 64 recording sites, whose performance is evaluated by the metrics of noise, spike amplitude, and spike detection probability. On average, we find no significant effect of device geometry on spike amplitude and detection probability but we find significant differences among individual experiments, with the likelihood of detecting spikes varying by a factor of approximately three across trials.", "date": "2012-10-15", "date_type": "published", "publication": "Journal of Neuroscience Methods", "volume": "211", "number": "1", "publisher": "Elsevier", "pagerange": "22-30", "id_number": "CaltechAUTHORS:20121217-082045717", "issn": "0165-0270", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20121217-082045717", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Eli and Edythe Broad Foundation" }, { "agency": "Della Martin Fund for Discoveries in Mental Illness" }, { "agency": "NIH", "grant_number": "DA-17279" }, { "agency": "NIH", "grant_number": "AG-033954" } ] }, "doi": "10.1016/j.jneumeth.2012.08.005", "pmcid": "PMC3473102", "primary_object": { "basename": "nihms-401238.pdf", "url": "https://authors.library.caltech.edu/records/nyy8d-kxg21/files/nihms-401238.pdf" }, "resource_type": "article", "pub_year": "2012", "author_list": "Scott, Kimberly M.; Du, Jiangang; et el." }, { "id": "https://authors.library.caltech.edu/records/cv21r-8kd26", "eprint_id": 35367, "eprint_status": "archive", "datestamp": "2023-08-19 12:58:39", "lastmod": "2023-10-20 16:09:07", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Insights into the Neurobiology of the Nicotinic Cholinergic System and Nicotine Addiction from Mice Expressing Nicotinic Receptors Harboring Gain-of-Function Mutations", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2012 by The American Society for Pharmacology and Experimental Therapeutics.\nPublished online before print August 10, 2012.\n\nThis work was supported by the National Institutes of Health National\nInstitute on Drug Abuse [Grants DA030396, DA19335,\nDA17279]; the National Institutes of Health National Institute on Aging\n[Grant AG33954]; the National Institutes of Health National Institute\nof Mental Health [Grant MH86383]; and the University of California\nTobacco Related Disease Research Program [Grant 17RT-0127].\nAuthorship Contributions:\nWrote or contributed to the writing of the manuscript: Drenan and\nLester.", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are ligand-gated, cation-selective ion channels expressed throughout the brain. Although these channels have been investigated for several decades, it is still challenging 1) to identify the important nAChR subunits in cholinergic transmission and nicotine dependence and 2) to develop nAChR subtype-specific ligands. To overcome these challenges, we and others have studied mice expressing mutant, gain-of-function nAChR subunits. In this review, we discuss this research approach and the results it has yielded to date. Gain-of-function mutations, including those in nAChR subunits, provide an approach that is complementary to loss-of-function studies such as gene knockouts; the former allows one to answer questions of sufficiency and the latter addresses questions of necessity. Mutant mice expressing gain-of-function nAChR subunits are commonly produced using traditional gene targeting in embryonic stem cells, but novel approaches such as bacterial artificial chromosome transgenesis have yielded important insights as well. \u03b17 nAChRs were the first nAChRs to be targeted with a gain-of-function mutation, followed by a pair of \u03b14 nAChR gain-of-function mutant mice. These \u03b14 nAChR gain-of-function mice (\u03b14 L9\u2032S mice, followed by \u03b14 L9\u2032A mice) provided an important system to probe \u03b14 nAChR function in vivo, particularly in the dopamine reward system. \u03b16 nAChR gain-of-function mice provided the first robust system allowing specific manipulation of this receptor subtype. Other targeted mutations in various nAChR subunits have also been produced and have yielded important insights into nicotinic cholinergic biology. As nAChR research advances and more details associated with nAChR expression and function emerge, we expect that existing and new mouse lines expressing gain-of-function nAChR subunits will continue to provide new insights.", "date": "2012-10", "date_type": "published", "publication": "Pharmacological Reviews", "volume": "64", "number": "4", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "869-879", "id_number": "CaltechAUTHORS:20121108-131347078", "issn": "0031-6997", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20121108-131347078", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA030396" }, { "agency": "NIH", "grant_number": "DA19335" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "AG33954" }, { "agency": "NIH", "grant_number": "MH86383" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "17RT-0127" } ] }, "doi": "10.1124/pr.111.004671", "pmcid": "PMC3462994", "resource_type": "article", "pub_year": "2012", "author_list": "Drenan, Ryan M. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/jm7ym-p7c66", "eprint_id": 35672, "eprint_status": "archive", "datestamp": "2023-08-19 12:59:31", "lastmod": "2023-10-20 20:28:12", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Duffy-N-H", "name": { "family": "Duffy", "given": "Noah H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Ondansetron and Granisetron Binding Orientation in the 5\u2011HT_3 \n Receptor Determined by Unnatural Amino Acid Mutagenesis", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2012 American Chemical Society. \n\nReceived: May 17, 2012. Accepted: August 8, 2012. Publication Date (Web): August 8, 2012. \n\nWe thank S. Lummis for helpful discussions and input. This\nwork was supported by the National Institutes of Health (NS\n34407 and DA19375). \n\nThe authors declare no competing financial interest.\n\nAccepted Version - nihms401305.pdf
", "abstract": "The serotonin type 3 receptor (5-HT_3R) is a ligand-gated ion channel found in the central and peripheral nervous systems. The 5-HT_3R is a therapeutic target, and the clinically available drugs ondansetron and granisetron inhibit receptor activity. Their inhibitory action is through competitive binding to the native ligand binding site, although the binding orientation of the drugs at the receptor has been a matter of debate. Here we heterologously express mouse 5-HT_3A receptors in Xenopus oocytes and use unnatural amino acid mutagenesis to establish a cation-\u03c0 interaction for both ondansetron and granisetron to tryptophan 183 in the ligand binding pocket. This cation-\u03c0 interaction establishes a binding orientation for both ondansetron and granisetron within the binding pocket.", "date": "2012-10", "date_type": "published", "publication": "ACS Chemical Biology", "volume": "7", "number": "10", "publisher": "American Chemical Society", "pagerange": "1738-1745", "id_number": "CaltechAUTHORS:20121127-104931725", "issn": "1554-8929", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20121127-104931725", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "DA19375" } ] }, "doi": "10.1021/cb300246j", "pmcid": "PMC3477246", "primary_object": { "basename": "nihms401305.pdf", "url": "https://authors.library.caltech.edu/records/jm7ym-p7c66/files/nihms401305.pdf" }, "resource_type": "article", "pub_year": "2012", "author_list": "Duffy, Noah H.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/g3rks-c4x36", "eprint_id": 35220, "eprint_status": "archive", "datestamp": "2023-08-19 12:41:52", "lastmod": "2023-10-20 15:51:47", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Daeffler-K-N-M", "name": { "family": "Daeffler", "given": "Kristina N.-M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Functionally Important Aromatic\u2013Aromatic and Sulfur\u2212\u03c0 Interactions in the D2 Dopamine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2012 American Chemical Society. \n\nReceived: May 10, 2012; Published: August 16, 2012; Published In Issue: September 12, 2012; Article ASAP: August 31, 2012. \n\nThis work was supported by the National Institutes of Health\ngrant GM081662. \n\nThe authors declare no competing financial interest.\n\nAccepted Version - nihms405177.pdf
Supplemental Material - ja304560x_si_001.pdf
Supplemental Material - ja304560x_si_002.pdf
", "abstract": "The recently published crystal structure of the D3 dopamine receptor shows a tightly packed region of aromatic residues on helices 5 and 6 in the space bridging the binding site and what is thought to be the origin of intracellular helical motion. This highly conserved region also makes contacts with residues on helix 3, and here we use double mutant cycle analysis and unnatural amino acid mutagenesis to probe the functional role of several residues in this region of the closely related D2 dopamine receptor. Of the eight mutant pairs examined, all show significant functional coupling (\u03a9 > 2), with the largest coupling coefficients observed between residues on different helices, C3.36/W6.48, T3.37/S5.46, and F5.47/F6.52. Additionally, three aromatic residues examined, F5.47, Y5.48, and F5.51, show consistent trends upon progressive fluorination of the aromatic side chain. These trends are indicative of a functionally important electrostatic interaction with the face of the aromatic residue examined, which is likely attributed to aromatic\u2013aromatic interactions between residues in this microdomain. We also propose that the previously determined fluorination trend at W6.48 is likely due to a sulfur\u2212\u03c0 interaction with the side chain of C3.36. We conclude that these residues form a tightly packed structural microdomain that connects helices 3, 5, and 6, thus forming a barrier that prevents dopamine from binding further toward the intracellular surface. Upon activation, these residues likely do not change their relative conformation, but rather act to translate agonist binding at the extracellular surface into the large intracellular movements that characterize receptor activation.", "date": "2012-09-12", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "134", "number": "36", "publisher": "American Chemical Society", "pagerange": "14890-14896", "id_number": "CaltechAUTHORS:20121031-153603159", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20121031-153603159", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM081662" } ] }, "doi": "10.1021/ja304560x", "pmcid": "PMC3461201", "primary_object": { "basename": "nihms405177.pdf", "url": "https://authors.library.caltech.edu/records/g3rks-c4x36/files/nihms405177.pdf" }, "related_objects": [ { "basename": "ja304560x_si_001.pdf", "url": "https://authors.library.caltech.edu/records/g3rks-c4x36/files/ja304560x_si_001.pdf" }, { "basename": "ja304560x_si_002.pdf", "url": "https://authors.library.caltech.edu/records/g3rks-c4x36/files/ja304560x_si_002.pdf" } ], "resource_type": "article", "pub_year": "2012", "author_list": "Daeffler, Kristina N.-M.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/0xvm2-40078", "eprint_id": 33950, "eprint_status": "archive", "datestamp": "2023-08-19 12:26:53", "lastmod": "2023-10-18 21:39:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Liu-Cambrian-Y", "name": { "family": "Liu", "given": "Cambrian Y." } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Fraser-S-E", "name": { "family": "Fraser", "given": "Scott E." }, "orcid": "0000-0002-5377-0223" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Koos-D-S", "name": { "family": "Koos", "given": "David S." } } ] }, "title": "Electrophysiological characterization of Grueneberg ganglion olfactory neurons: spontaneous firing, sodium conductance, and\n hyperpolarization-activated currents", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2012 American Physiological Society. \n\nSubmitted 5 October 2011. Accepted in final form 29 May 2012. \nPublished online before print May 30, 2012. This information is current as of September 7, 2012. \n\nWe thank J. Gutierrez, A. R. Douglas, and S. M. M. Alaniz for animal care and husbandry. This project was supported by grants from the National Institutes of Health and the National Science Foundation. No conflicts of interest, financial or otherwise, are declared by the authors. Author contributions: C.Y.L., H.A.L., and D.S.K. conception and design of research; C.Y.L. and C.X. performed experiments; C.Y.L. and C.X. analyzed data; C.Y.L., C.X., and H.A.L. interpreted results of experiments; C.Y.L. prepared figures; C.Y.L. drafted manuscript; C.Y.L., C.X., S.E.F., H.A.L., and D.S.K. edited and revised manuscript; S.E.F. and D.S.K. approved final version of manuscript.", "abstract": "Mammals rely on their acute olfactory sense for their survival. The most anterior olfactory subsystem in the nose, the Grueneberg ganglion (GG), plays a role in detecting alarm pheromone, cold, and urinary compounds. GG neurons respond homogeneously to these stimuli with increases in intracellular [Ca^(2+)] or transcription of immediate-early genes. In this electrophysiological study, we used patch clamp techniques to characterize the membrane properties of GG neurons. Our results offer evidence of functional heterogeneity in the GG. GG neurons fire spontaneously and independently in several stable patterns, including phasic and repetitive single-spike modes of discharge. Whole-cell recordings demonstrated two distinct voltage-gated fast-inactivating Na^+ currents with different steady-state voltage dependencies and different sensitivities to tetrodotoxin. Hodgkin-Huxley simulations showed that these Na^+ currents confer dual mechanisms of action potential generation and contribute to different firing patterns. Additionally, GG neurons exhibited hyperpolarization-activated inward currents that modulated spontaneous firing in vitro. Thus, in GG neurons the heterogeneity of firing patterns is linked to the unusual repertoire of ionic currents. The membrane properties described here will aid the interpretation of chemosensory function in the GG.", "date": "2012-09", "date_type": "published", "publication": "Journal of Neurophysiology", "volume": "108", "number": "5", "publisher": "American Physiological Society", "pagerange": "1318-1334", "id_number": "CaltechAUTHORS:20120910-073630839", "issn": "0022-3077", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120910-073630839", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "NSF" } ] }, "pmcid": "PMC3544954", "resource_type": "article", "pub_year": "2012", "author_list": "Liu, Cambrian Y.; Xiao, Cheng; et el." }, { "id": "https://authors.library.caltech.edu/records/t37gq-68p43", "eprint_id": 34462, "eprint_status": "archive", "datestamp": "2023-09-14 19:33:53", "lastmod": "2023-10-23 20:53:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Shapiro-M-G", "name": { "family": "Shapiro", "given": "Mikhail G." }, "orcid": "0000-0002-0291-4215" }, { "id": "Frazier-S-J", "name": { "family": "Frazier", "given": "Shawnalea J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Unparalleled Control of Neural Activity Using Orthogonal Pharmacogenetics", "ispublished": "pub", "full_text_status": "public", "keywords": "Pharmacology; pharmacogenetics; neural control; systems neuroscience; brain interfaces; optogenetics", "note": "\u00a9 2012 American Chemical Society.\n\nReceived: May 4, 2012; Accepted: June 1, 2012; Published: June 1, 2012. \n\nThe authors declare no competing financial interest.\n\nPublished - cn300053q.pdf
", "abstract": "Studying the functional architecture of the brain requires technologies to precisely measure and perturb the activity of specific neural cells and circuits in live animals. Substantial progress has been made in recent years to develop and apply such tools. In particular, technologies that provide precise control of activity in genetically defined populations of neurons have enabled the study of causal relationships between and among neural circuit elements and behavioral outputs. Here, we review an important subset of such technologies, in which neurons are genetically engineered to respond to specific chemical ligands that have no interfering pharmacological effect in the central nervous system. A rapidly expanding set of these \"orthogonal pharmacogenetic\" tools provides a unique combination of genetic specificity, functional diversity, spatiotemporal precision, and potential for multiplexing. We review the main classes of orthogonal pharmacogenetic technologies, including neuroreceptors to control neuronal excitability, systems to control gene transcription and translation, and general constructs to control protein\u2013protein interactions, enzymatic function, and protein stability. We describe the key performance characteristics informing the use of these technologies in the brain, and potential directions for improvement and expansion of the orthogonal pharmacogenetics toolkit to enable more sophisticated systems neuroscience.", "date": "2012-08-15", "date_type": "published", "publication": "ACS Chemical Neuroscience", "volume": "3", "number": "8", "publisher": "American Chemical Society", "pagerange": "619-629", "id_number": "CaltechAUTHORS:20120926-114816752", "issn": "1948-7193", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120926-114816752", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1021/cn300053q", "pmcid": "PMC3419455", "primary_object": { "basename": "cn300053q.pdf", "url": "https://authors.library.caltech.edu/records/t37gq-68p43/files/cn300053q.pdf" }, "resource_type": "article", "pub_year": "2012", "author_list": "Shapiro, Mikhail G.; Frazier, Shawnalea J.; et el." }, { "id": "https://authors.library.caltech.edu/records/7nsva-c6493", "eprint_id": 34315, "eprint_status": "archive", "datestamp": "2023-08-19 12:01:26", "lastmod": "2023-10-19 14:49:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Miwa-J-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Walz-A", "name": { "family": "Walz", "given": "Andreas" } } ] }, "title": "Optimizing Cholinergic Tone Through Lynx Modulators of Nicotinic Receptors: Implications for Plasticity and Nicotine Addiction", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2012 International Union of the Physiological Sciences/American Physiological Society.\n\n\nWe thank Sharon Grady and Rell L. Parker for helpful\ndiscussions and Pauline Ku and Wesley Chen for help with\nfigures. This work was supported by funds from Tobacco-Related Disease Research Program of the University of California, Grant No. TRDRP19KT-0032 for J. M. Miwa; 5R43MH083416-02 and 1R43MH094004-01 for A. Walz; and R01 AG-033954-01 for H. A. Lester and J. M. Miwa. J. M. Miwa holds shares in Ophidion. A. Walz holds shares in Ophidion. Ophidion pursues the development of cognitive enhancement therapies related to the subject matter. Author contributions: J.M.M. analyzed data; J.M.M. interpreted\nresults of experiments; J.M.M. and A.W. prepared\nfigures; J.M.M. drafted manuscript; J.M.M., A.W.,\nand H.A.L. edited and revised manuscript; J.M.M., A.W.,\nand H.A.L. approved final version of manuscript.", "abstract": "The cholinergic system underlies both adaptive (learning and memory) and nonadaptive (addiction and dependency) behavioral changes through its ability to shape and regulate plasticity. Protein modulators such as lynx family members can fine tune the activity of the cholinergic system and contribute to the graded response of the cholinergic system, stabilizing neural circuitry through direct interaction with nicotinic receptors. Release of this molecular brake can unmask cholinergic-dependent mechanisms in the brain. Lynx proteins have the potential to provide top-down control over plasticity mechanisms, including addictive propensity. If this is indeed the case, then, what regulates the regulator? Transcriptional changes of lynx genes in response to pharmacological, physiological, and pathological alterations are explored in this review.", "date": "2012-08-01", "date_type": "published", "publication": "Physiology", "volume": "27", "number": "4", "publisher": "American Physiological Society", "pagerange": "187-199", "id_number": "CaltechAUTHORS:20120924-131621260", "issn": "1548-9221", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120924-131621260", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "TRDRP19KT-0032" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "5R43MH083416-02" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "1R43MH094004-01" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "R01 AG-033954-01" } ] }, "doi": "10.1152/physiol.00002.2012", "resource_type": "article", "pub_year": "2012", "author_list": "Miwa, Julie M.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/mwg10-kyd21", "eprint_id": 33941, "eprint_status": "archive", "datestamp": "2023-08-22 06:17:23", "lastmod": "2023-10-18 21:39:00", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Richards-C-I", "name": { "family": "Richards", "given": "Christopher I." } }, { "id": "Dilworth-C", "name": { "family": "Dilworth", "given": "Crystal" } }, { "id": "Moss-F-J", "name": { "family": "Moss", "given": "Fraser J." }, "orcid": "0000-0002-8519-6991" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "F\u00f6rster Resonance Energy Transfer (FRET) Correlates of Altered Subunit Stoichiometry in Cys-Loop Receptors, Exemplified by Nicotinic \u03b14\u03b22", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotine; cytisine; NFRET; nicotine addiction; Parkinson's disease; ion channels", "note": "\u00a9 2012 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article\ndistributed under the terms and conditions of the Creative Commons Attribution license\n(http://creativecommons.org/licenses/by/3.0/).\n\nReceived: 30 May 2012; in revised form: 2 July 2012; Accepted: 5 July 2012; Published: 10 August 2012.\nWe thank Kimberly Scott for discussions. Supported by grants from the U.S. National Institutes of\nHealth (NS-11756, NS-34407, AG-033954), Targacept Inc., Louis and Janet Fletcher, the Michael J.\nFox Foundation, the California Tobacco-Related Disease Research Program postdoctoral fellowship\n(18FT-0066 to R.S.), a Beckman Institute Fellowship (C.I.R) and an NIH Kirschstein National\nResearch Service Award (DA030877 to C.I.R).\n\nPublished - Srinivasan2012p19376Int_J_Mol_Sci.pdf
Supplemental Material - ijms-13-10022-s001.pdf
", "abstract": "We provide a theory for employing F\u00f6rster resonance energy transfer (FRET)\nmeasurements to determine altered heteropentameric ion channel stoichiometries in\nintracellular compartments of living cells. We simulate FRET within nicotinic receptors\n(nAChRs) whose \u03b14 and \u03b22 subunits contain acceptor and donor fluorescent protein\nmoieties, respectively, within the cytoplasmic loops. We predict FRET and normalized\nFRET (NFRET) for the two predominant stoichiometries, (\u03b14)3(\u03b22)2 vs. (\u03b14)2(\u03b22)3.\nStudying the ratio between FRET or NFRET for the two stoichiometries, minimizes\ndistortions due to various photophysical uncertainties. Within a range of assumptions\nconcerning the distance between fluorophores, deviations from plane pentameric geometry,\nand other asymmetries, the predicted FRET and NFRET for (\u03b14)3(\u03b22)2 exceeds that of\n(\u03b14)2(\u03b22)3. The simulations account for published data on transfected Neuro2a cells in\nwhich \u03b14\u03b22 stoichiometries were manipulated by varying fluorescent subunit cDNA ratios:\nNFRET decreased monotonically from (\u03b14)3(\u03b22)2 stoichiometry to mostly (\u03b14)2(\u03b22)3. The\nsimulations also account for previous macroscopic and single-channel observations that\npharmacological chaperoning by nicotine and cytisine increase the (\u03b14)2(\u03b22)3 and\n(\u03b14)3(\u03b22)2 populations, respectively. We also analyze sources of variability. NFRET-based monitoring of changes in subunit stoichiometry can contribute usefully to studies on\nCys-loop receptors.", "date": "2012-08", "date_type": "published", "publication": "International Journal of Molecular Sciences", "volume": "13", "number": "8", "publisher": "MDPI", "pagerange": "10022-10040", "id_number": "CaltechAUTHORS:20120907-141513430", "issn": "1422-0067", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120907-141513430", "rights": "This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "AG-033954" }, { "agency": "Targacept Inc." }, { "agency": "Louis and Janet Fletcher" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "18FT-0066" }, { "agency": "Caltech Beckman Institute" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "DA030877" } ] }, "doi": "10.3390/ijms130810022", "pmcid": "PMC3431844", "primary_object": { "basename": "Srinivasan2012p19376Int_J_Mol_Sci.pdf", "url": "https://authors.library.caltech.edu/records/mwg10-kyd21/files/Srinivasan2012p19376Int_J_Mol_Sci.pdf" }, "related_objects": [ { "basename": "ijms-13-10022-s001.pdf", "url": "https://authors.library.caltech.edu/records/mwg10-kyd21/files/ijms-13-10022-s001.pdf" } ], "resource_type": "article", "pub_year": "2012", "author_list": "Srinivasan, Rahul; Richards, Christopher I.; et el." }, { "id": "https://authors.library.caltech.edu/records/xrnsz-e3q23", "eprint_id": 33814, "eprint_status": "archive", "datestamp": "2023-08-19 11:47:19", "lastmod": "2023-10-18 20:41:46", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "Engle-S-E", "name": { "family": "Engle", "given": "Staci E." } }, { "id": "Kim-Mi-Ran", "name": { "family": "Kim", "given": "Mi Ran" } }, { "id": "O'Neill-H-C", "name": { "family": "O'Neill", "given": "Heidi C." } }, { "id": "Wageman-C-R", "name": { "family": "Wageman", "given": "Charles R." } }, { "id": "Patzlaff-N-E", "name": { "family": "Patzlaff", "given": "Natalie E." } }, { "id": "Wang-Ying", "name": { "family": "Wang", "given": "Ying" } }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "McIntosh-J-M", "name": { "family": "McIntosh", "given": "J. Michael" } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" } ] }, "title": "\u03b16* Nicotinic Acetylcholine Receptor Expression and Function in a Visual Salience Circuit", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2012 The Authors. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). \n\nReceived Jan. 2, 2012; revised April 14, 2012; accepted May 14, 2012.\n\nAuthor contributions: E.D.W.M., H.C.O., S.R.G., M.J.M., and R.M.D. designed research; E.D.W.M., S.E.E., M.R.K.,\nH.C.O., C.R.W., N.E.P., Y.W., S.R.G., M.J.M., and R.M.D. performed research; Y.W. contributed unpublished reagents/\nanalytic tools; E.D.W.M., S.E.E., M.R.K., H.C.O., Y.W., S.R.G., M.J.M., H.A.L., and R.M.D. analyzed data; E.D.W.M.,\nS.R.G., J.M.M., M.J.M., H.A.L., and R.M.D. wrote the paper.\nThis work was supported by NIH Grants DA030396, DA17279, DA12242, DA015663, DA03194, MH53631, and GM48677. Thanks to the Purdue Histology and Phenotyping Laboratory (Purdue University, School of Veterinary Medicine) for technical assistance with histology procedures. The authors declare no competing financial interests.\n\nPublished - Mackey2012p19334J_Neurosci.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) containing \u03b16 subunits are expressed in only a few brain areas, including midbrain dopamine (DA) neurons, noradrenergic neurons of the locus ceruleus, and retinal ganglion cells. To better understand the regional and subcellular expression pattern of \u03b16-containing nAChRs, we created and studied transgenic mice expressing a variant \u03b16 subunit with green fluorescent protein (GFP) fused in-frame in the M3-M4 intracellular loop. In \u03b16-GFP transgenic mice, \u03b16-dependent synaptosomal DA release and radioligand binding experiments confirmed correct expression and function in vivo. In addition to strong \u03b16* nAChR expression in glutamatergic retinal axons, which terminate in superficial superior colliculus (sSC), we also found \u03b16 subunit expression in a subset of GABAergic cell bodies in this brain area. In patch-clamp recordings from sSC neurons in brain slices from mice expressing hypersensitive \u03b16* nAChRs, we confirmed functional, postsynaptic \u03b16* nAChR expression. Further, sSC GABAergic neurons expressing \u03b16* nAChRs exhibit a tonic conductance mediated by standing activation of hypersensitive \u03b16* nAChRs by ACh. \u03b16* nAChRs also appear in a subpopulation of SC neurons in output layers. Finally, selective activation of \u03b16* nAChRs in vivo induced sSC neuronal activation as measured with c-Fos expression. Together, these results demonstrate that \u03b16* nAChRs are uniquely situated to mediate cholinergic modulation of glutamate and GABA release in SC. The SC has emerged as a potential key brain area responsible for transmitting short-latency salience signals to thalamus and midbrain DA neurons, and these results suggest that \u03b16* nAChRs may be important for nicotinic cholinergic sensitization of this pathway.", "date": "2012-07-25", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "32", "number": "30", "publisher": "Society for Neuroscience", "pagerange": "10226-10237", "id_number": "CaltechAUTHORS:20120904-085001085", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120904-085001085", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA030396" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "DA12242" }, { "agency": "NIH", "grant_number": "DA015663" }, { "agency": "NIH", "grant_number": "DA03194" }, { "agency": "NIH", "grant_number": "MH53631" }, { "agency": "NIH", "grant_number": "GM48677" } ] }, "doi": "10.1523/JNEUROSCI.0007-12.2012", "pmcid": "PMC3432940", "primary_object": { "basename": "Mackey2012p19334J_Neurosci.pdf", "url": "https://authors.library.caltech.edu/records/xrnsz-e3q23/files/Mackey2012p19334J_Neurosci.pdf" }, "resource_type": "article", "pub_year": "2012", "author_list": "Mackey, Elisha D. W.; Engle, Staci E.; et el." }, { "id": "https://authors.library.caltech.edu/records/y4b0p-tpb51", "eprint_id": 33518, "eprint_status": "archive", "datestamp": "2023-08-19 11:44:06", "lastmod": "2023-10-18 20:23:20", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Tavares-X-D-S", "name": { "family": "Tavares", "given": "Ximena Da Silva" } }, { "id": "Blum-A-P", "name": { "family": "Blum", "given": "Angela P." } }, { "id": "Nakamura-D-T", "name": { "family": "Nakamura", "given": "Darren T." } }, { "id": "Puskar-N-L", "name": { "family": "Puskar", "given": "Nyssa L." } }, { "id": "Shanata-J-A-P", "name": { "family": "Shanata", "given": "Jai A. P." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Variations in Binding Among Several Agonists at Two Stoichiometries of the Neuronal, \u03b14\u03b22 Nicotinic Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2012 American Chemical Society.\n\nReceived: February 14, 2012; Published: June 20, 2012.\n\nThis work was supported by the National Institutes of Health\n[Grants NS34407 NS11756] and by the California Tobacco-Related Disease Research Program of the University of\nCalifornia, Award 19XT-0102.\n\nPublished - Tavares2012p19275J_Am_Chem_Soc.pdf
Accepted Version - nihms-389562.pdf
Supplemental Material - ja3011379_si_001.pdf
", "abstract": "Drug-receptor binding interactions of four agonists, ACh, nicotine, and the smoking cessation compounds varenicline (Chantix) and cytisine (Tabex), have been evaluated at both the 2:3 and 3:2 stoichiometries of the \u03b14\u03b22 nicotinic acetylcholine receptor (nAChR). Previous studies have established that unnatural amino acid mutagenesis can probe three key binding interactions at the nAChR: a cation\u2212\u03c0 interaction, and two hydrogen-bonding interactions to the protein backbone of the receptor. We find that all drugs make a cation\u2212\u03c0 interaction to TrpB of the receptor. All drugs except ACh, which lacks an N^(+)H group, make a hydrogen bond to a backbone carbonyl, and ACh and nicotine behave similarly in acting as a hydrogen-bond acceptor. However, varenicline is not a hydrogen-bond acceptor to the backbone NH that interacts strongly with the other three compounds considered. In addition, we see interesting variations in hydrogen bonding interactions with cytisine that provide a rationalization for the stoichiometry selectivity seen with this compound.", "date": "2012-07-18", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "134", "number": "28", "publisher": "American Chemical Society", "pagerange": "11474-11480", "id_number": "CaltechAUTHORS:20120824-131040943", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120824-131040943", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19XT-0102" } ] }, "doi": "10.1021/ja3011379", "pmcid": "PMC3399941", "primary_object": { "basename": "Tavares2012p19275J_Am_Chem_Soc.pdf", "url": "https://authors.library.caltech.edu/records/y4b0p-tpb51/files/Tavares2012p19275J_Am_Chem_Soc.pdf" }, "related_objects": [ { "basename": "ja3011379_si_001.pdf", "url": "https://authors.library.caltech.edu/records/y4b0p-tpb51/files/ja3011379_si_001.pdf" }, { "basename": "nihms-389562.pdf", "url": "https://authors.library.caltech.edu/records/y4b0p-tpb51/files/nihms-389562.pdf" } ], "resource_type": "article", "pub_year": "2012", "author_list": "Tavares, Ximena Da Silva; Blum, Angela P.; et el." }, { "id": "https://authors.library.caltech.edu/records/2j93j-84002", "eprint_id": 33128, "eprint_status": "archive", "datestamp": "2023-08-22 06:04:49", "lastmod": "2023-10-18 18:49:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Richards-C-I", "name": { "family": "Richards", "given": "Christopher I." } }, { "id": "Luong-Khai", "name": { "family": "Luong", "given": "Khai" } }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Turner-S-W", "name": { "family": "Turner", "given": "Stephen W." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Korlach-J", "name": { "family": "Korlach", "given": "Jonas" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Live-Cell Imaging of Single Receptor Composition Using Zero-Mode Waveguide Nanostructures", "ispublished": "pub", "full_text_status": "public", "keywords": "Membrane receptors; nanostructures; single-molecule imaging; nicotinic receptors; zero-mode waveguide", "note": "\u00a9 2012 American Chemical Society. Received: April 20, 2012.\nRevised: June 2, 2012. Publication Date (Web): June 5, 2012. We thank A. Fernandez and M. Boitano for assistance in\npreparing ZMW arrays. Supported by a Beckman Fellowship\nand an NIH NRSA to C.I.R., by a California Tobacco-Related\nDisease Research Fellowship to R.S., and by the US National\nInstitutes of Health (NS34407).\n\nPublished - Richards2012p19101Nano_Lett.pdf
Accepted Version - nihms-383248.pdf
Supplemental Material - nl301480h_si_001.pdf
", "abstract": "We exploit the optical and spatial features of subwavelength nanostructures to examine individual receptors on the plasma membrane of living cells. Receptors were sequestered in portions of the membrane projected into zero-mode waveguides. Using single-step photobleaching of green fluorescent protein incorporated into individual subunits, the resulting spatial isolation was used to measure subunit stoichiometry in \u03b14\u03b24 and \u03b14\u03b22 nicotinic acetylcholine and P2X2 ATP receptors. We also show that nicotine and cytisine have differential effects on \u03b14\u03b22 stoichiometry.", "date": "2012-07", "date_type": "published", "publication": "Nano Letters", "volume": "12", "number": "7", "publisher": "American Chemical Society", "pagerange": "3690-3694", "id_number": "CaltechAUTHORS:20120813-111734004", "issn": "1530-6984", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120813-111734004", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Arnold and Mabel Beckman Foundation" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "NIH", "grant_number": "NS34407" } ] }, "doi": "10.1021/nl301480h", "pmcid": "PMC3397148", "primary_object": { "basename": "Richards2012p19101Nano_Lett.pdf", "url": "https://authors.library.caltech.edu/records/2j93j-84002/files/Richards2012p19101Nano_Lett.pdf" }, "related_objects": [ { "basename": "nihms-383248.pdf", "url": "https://authors.library.caltech.edu/records/2j93j-84002/files/nihms-383248.pdf" }, { "basename": "nl301480h_si_001.pdf", "url": "https://authors.library.caltech.edu/records/2j93j-84002/files/nl301480h_si_001.pdf" } ], "resource_type": "article", "pub_year": "2012", "author_list": "Richards, Christopher I.; Luong, Khai; et el." }, { "id": "https://authors.library.caltech.edu/records/k02z9-3hw15", "eprint_id": 31897, "eprint_status": "archive", "datestamp": "2023-08-19 10:59:14", "lastmod": "2023-10-17 21:40:59", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Puskar-N-L", "name": { "family": "Puskar", "given": "Nyssa L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Probing the Effects of Residues Located Outside the Agonist Binding Site on Drug-Receptor Selectivity in the Nicotinic Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2012 American Chemical Society.\n\nPublished In Issue May 18, 2012; Article ASAP: February 14, 2012; Just Accepted Manuscript: February 01, 2012; Received: October 31, 2011; Accepted: February 01, 2012.\n\nThis work was supported by the National Institutes of Health\n(NS 34407 and NS 11756). We also thank a referee for\npointing out the sequence differences between the Limanea and Aplysia forms of AChBP.\n\nAccepted Version - nihms355205.pdf
Supplemental Material - cb200448j_si_001.pdf
", "abstract": "The nicotinic acetylcholine receptors (nAChRs) are a family of closely related but pharmacologically distinct neurotransmitter-gated ion channels. They are therapeutic targets for a wide range of neurological disorders, and a key issue in drug development is selective targeting among the more than 20 subtypes of nAChRs that are known. The present work evaluates a proposed hydrogen bonding interaction involving a residue known as the \"loop B glycine\" that distinguishes receptors that are highly responsive to ACh and nicotine from those that are much less so. We have performed structure\u2013function studies on the loop B site, including unnatural amino acid mutagenesis, in three different nAChR subtypes and found that the correlation between agonist potency and this residue is strong. Low potency receptor subtypes have a glycine at this key site, and mutation to a residue with a side chain converts a low potency receptor to a high potency receptor. Innately high potency receptors have a lysine at the loop B site and show a decrease in potency for the reverse mutation (i.e., introducing a glycine). This residue lies outside of the agonist binding site, and studies of other residues at the agonist binding site show that the details of how changes at the loop B glycine site impact agonist potency vary for differing receptor subtypes. This suggests a model in which the loop B residue influences the global shape of the agonist binding site rather than modulating any specific interaction.", "date": "2012-05-18", "date_type": "published", "publication": "ACS Chemical Biology", "volume": "7", "number": "5", "publisher": "American Chemical Society", "pagerange": "841-846", "id_number": "CaltechAUTHORS:20120613-133532352", "issn": "1554-8929", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120613-133532352", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1021/cb200448j", "pmcid": "PMC3356501", "primary_object": { "basename": "cb200448j_si_001.pdf", "url": "https://authors.library.caltech.edu/records/k02z9-3hw15/files/cb200448j_si_001.pdf" }, "related_objects": [ { "basename": "nihms355205.pdf", "url": "https://authors.library.caltech.edu/records/k02z9-3hw15/files/nihms355205.pdf" } ], "resource_type": "article", "pub_year": "2012", "author_list": "Puskar, Nyssa L.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/4fmkj-dzr64", "eprint_id": 31692, "eprint_status": "archive", "datestamp": "2023-08-19 10:04:08", "lastmod": "2023-10-17 18:46:55", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Richards-C-I", "name": { "family": "Richards", "given": "Christopher I." } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Rhee-Doreen", "name": { "family": "Rhee", "given": "Doreen" } }, { "id": "Pantoja-R", "name": { "family": "Pantoja", "given": "Rigo" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Miwa-Julie-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Pharmacological Chaperoning of Nicotinic Acetylcholine Receptors Reduces the Endoplasmic Reticulum Stress Response", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2012 The American Society for Pharmacology and Experimental Therapeutics. \n\nReceived January 13, 2012; accepted February 28, 2012. Published online before print February 29, 2012. \n\nThis work was supported by the National Institutes of Health National Institute of Neurological Disorders and Stroke [Grant NS11756]; the National Institutes of Health National Institute on Aging [Grant AG033954]; National Institutes of Health National Institute on Drug Abuse Kirschstein National Research Service Award [Grant DA030877] (to C.I.R.); Targacept; Louis and Janet Fletcher; the Michael J. Fox Foundation; a California Tobacco-Related Disease Research Program postdoctoral fellowship [Grant 18FT-0066] (to R.S.); a Beckman Institute fellowship (to C.I.R.); and a California Tobacco-Related Disease Research Program New Investigator Award [Grant 19KT-0032] (to J.M.M.). \n\nWe thank Johannes Schwarz for discussion, Elisha D. W. Mackey\nfor assistance with cloning, and Sheri McKinney for assistance with primary neuronal cultures. \n\nAuthorship Contributions: Participated in research design: Srinivasan, Richards, Pantoja, and Lester.\nConducted experiments: Srinivasan, Richards, Xiao, Rhee, and\nPantoja.\nContributed new reagents or analytic tools: Srinivasan, Richards,\nand Miwa.\nPerformed data analysis: Srinivasan, Richards, Xiao, and Lester.\nWrote or contributed to the writing of the manuscript: Srinivasan,\nRichards, Dougherty, and Lester.\n\nSupplemental Material - Supplementary_figure_1.pdf
", "abstract": "We report the first observation that endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) can decrease when a central nervous system drug acts as an intracellular pharmacological chaperone for its classic receptor. Transient expression of \u03b14\u03b22 nicotinic receptors (nAChRs) in Neuro-2a cells induced the nuclear translocation of activating transcription factor 6 (ATF6), which is part of the UPR. Cells were exposed for 48 h to the full agonist nicotine, the partial agonist cytisine, or the competitive antagonist dihydro-\u03b2-erythroidine; we also tested mutant nAChRs that readily exit the ER. Each of these four manipulations increased Sec24D-enhanced green fluorescent protein fluorescence of condensed ER exit sites and attenuated translocation of ATF6-enhanced green fluorescent protein to the nucleus. However, we found no correlation among the manipulations regarding other tested parameters [i.e., changes in nAChR stoichiometry (\u03b14_2\u03b22_3 versus \u03b14_3\u03b22_2), changes in ER and trans-Golgi structures, or the degree of nAChR up-regulation at the plasma membrane]. The four manipulations activated 0 to 0.4% of nAChRs, which shows that activation of the nAChR channel did not underlie the reduced ER stress. Nicotine also attenuated endogenously expressed ATF6 translocation and phosphorylation of eukaryotic initiation factor 2\u03b1 in mouse cortical neurons transfected with \u03b14\u03b22 nAChRs. We conclude that, when nicotine accelerates ER export of \u03b14\u03b22 nAChRs, this suppresses ER stress and the UPR. Suppression of a sustained UPR may explain the apparent neuroprotective effect that causes the inverse correlation between a person's history of tobacco use and susceptibility to developing Parkinson's disease. This suggests a novel mechanism for neuroprotection by nicotine.", "date": "2012-02-29", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "81", "number": "6", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "759-769", "id_number": "CaltechAUTHORS:20120530-081520292", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120530-081520292", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "AG0033954" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "DA030877" }, { "agency": "Targacept" }, { "agency": "Louis and Janet Fletcher" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "18FT-0066" }, { "agency": "Caltech Beckman Institute" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19KT-0032" }, { "agency": "National Institute of Neurological Disorders and Stroke (NINDS)" }, { "agency": "National Institute on Aging" }, { "agency": "National Institute on Drug Abuse" } ] }, "doi": "10.1124/mol.112.077792", "pmcid": "PMC3362896", "primary_object": { "basename": "Supplementary_figure_1.pdf", "url": "https://authors.library.caltech.edu/records/4fmkj-dzr64/files/Supplementary_figure_1.pdf" }, "resource_type": "article", "pub_year": "2012", "author_list": "Srinivasan, Rahul; Richards, Christopher I.; et el." }, { "id": "https://authors.library.caltech.edu/records/6qavs-7qf18", "eprint_id": 29287, "eprint_status": "archive", "datestamp": "2023-08-19 09:44:58", "lastmod": "2023-10-20 21:56:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Murray-T-A", "name": { "family": "Murray", "given": "Teresa A." } }, { "id": "Bertrand-D", "name": { "family": "Bertrand", "given": "Daniel" } }, { "id": "Papke-R-L", "name": { "family": "Papke", "given": "Roger L." } }, { "id": "George-A-A", "name": { "family": "George", "given": "Andrew A." } }, { "id": "Pantoja-R", "name": { "family": "Pantoja", "given": "Rigo" } }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Liu-Qiang", "name": { "family": "Liu", "given": "Qiang" } }, { "id": "Wu-Jie", "name": { "family": "Wu", "given": "Jie" } }, { "id": "Whiteaker-P", "name": { "family": "Whiteaker", "given": "Paul" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Lukas-R-J", "name": { "family": "Lukas", "given": "Ronald J." } } ] }, "title": "\u03b17\u03b22 Nicotinic Acetylcholine Receptors Assemble, Function,\n and Are Activated Primarily via Their \u03b17-\u03b17 Interfaces", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2012 The American Society for Pharmacology and Experimental Therapeutics. \n\nReceived June 12, 2011; accepted October 28, 2011. \n\nThis work was supported by the National Institutes of Health National Institute on Drug Abuse [Grants DA015389, DA027070, DA012242]; the National Institutes of Health National Institute of Neurological Diseases and Stroke [Grant NS11756]; the National Institutes of Health National Institute of Mental Health [Grant MH086383]; the National Institutes of Health National\nInstitute of General Medical Sciences [Grant GM057481]; a US National Science Foundation Graduate Research Fellowship; the Barrow Neurological Foundation; a Catholic Healthcare West SEED Grant; the Biodesign Institute at Arizona State University, the Swiss National Science Foundation; the EC Neurocypres Grant; and the California Tobacco-Related Disease Research Program.\nAuthorship Contributions:\nParticipated in research design: Murray, Bertrand, Papke, George, Pantoja, Srinivasan, Liu, Wu, Whiteaker, Lester, and Lukas. Conducted experiments: Murray, Bertrand, Papke, George, and Liu. Performed data analysis: Murray, Bertrand, Papke, George, and\nLiu. Wrote or contributed to the writing of the manuscript: Murray, Bertrand, Papke, George, Liu, Whiteaker, Lester and Lukas.\n\nErratum - mol.111.074088err.pdf
", "abstract": "We investigated assembly and function of nicotinic acetylcholine receptors (nAChRs) composed of \u03b17 and \u03b22 subunits. We measured optical and electrophysiological properties of wild-type and mutant subunits expressed in cell lines and Xenopus laevis oocytes. Laser scanning confocal microscopy indicated that fluorescently tagged \u03b17 and \u03b22 subunits colocalize. F\u00f6rster resonance energy transfer between fluorescently tagged subunits strongly suggested that \u03b17 and \u03b22 subunits coassemble. Total internal reflection fluorescence microscopy revealed that assemblies localized to filopodia-like processes of SH-EP1 cells. Gain-of-function \u03b17 and \u03b22 subunits confirmed that these subunits coassemble within functional receptors. Moreover, \u03b17\u03b22 nAChRs composed of wild-type subunits or fluorescently tagged subunits had pharmacological properties similar to those of \u03b17 nAChRs, although amplitudes of \u03b17\u03b22 nAChR-mediated, agonist-evoked currents were generally \u223c2-fold lower than those for \u03b17 nAChRs. It is noteworthy that \u03b17\u03b22 nAChRs displayed sensitivity to low concentrations of the antagonist dihydro-\u03b2-erythroidine that was not observed for \u03b17 nAChRs at comparable concentrations. In addition, cysteine mutants revealed that the \u03b17-\u03b22 subunit interface does not bind ligand in a functionally productive manner, partly explaining lower \u03b17\u03b22 nAChR current amplitudes and challenges in identifying the function of native \u03b17\u03b22 nAChRs. On the basis of our findings, we have constructed a model predicting receptor function that is based on stoichiometry and position of \u03b22 subunits within the \u03b17\u03b22 nAChRs.", "date": "2012-02", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "81", "number": "2", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "175-188", "id_number": "CaltechAUTHORS:20120214-145000544", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120214-145000544", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA015389" }, { "agency": "NIH", "grant_number": "DA027070" }, { "agency": "NIH", "grant_number": "DA012242" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "MH086383" }, { "agency": "NIH", "grant_number": "GM057481" }, { "agency": "NSF Graduate Research Fellowship" }, { "agency": "Barrow Neurological Foundation" }, { "agency": "Catholic Heathcare West SEED Grant" }, { "agency": "Arizona State University Biodesign Institute" }, { "agency": "Swiss National Science Foundation (SNSF)" }, { "agency": "EC Neurocypres Grant" }, { "agency": "California Tobacco-Related Disease Research Program" } ] }, "doi": "10.1124/mol.111.074088", "pmcid": "PMC3263954", "primary_object": { "basename": "mol.111.074088err.pdf", "url": "https://authors.library.caltech.edu/records/6qavs-7qf18/files/mol.111.074088err.pdf" }, "resource_type": "article", "pub_year": "2012", "author_list": "Murray, Teresa A.; Bertrand, Daniel; et el." }, { "id": "https://authors.library.caltech.edu/records/rx81d-bgj35", "eprint_id": 29268, "eprint_status": "archive", "datestamp": "2023-08-22 04:41:34", "lastmod": "2023-10-24 22:03:21", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "B. N." } }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "E. D. W." } }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "S. R." } }, { "id": "Mckinney-S", "name": { "family": "Mckinney", "given": "S." } }, { "id": "Patzlaff-N-E", "name": { "family": "Patzlaff", "given": "N. E." } }, { "id": "Wageman-C-R", "name": { "family": "Wageman", "given": "C. R." } }, { "id": "Mcintosh-J-M", "name": { "family": "Mcintosh", "given": "J. M." } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "M. J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "R. M." }, "orcid": "0000-0002-8141-8577" } ] }, "title": "Nicotinic cholinergic mechanisms causing elevated dopamine release and abnormal locomotor behavior", "ispublished": "pub", "full_text_status": "public", "keywords": "dopamine; cholinergic; nicotinic; acetylcholine; transgenic; mouse", "note": "\u00a9 2011 IBRO. Published by Elsevier Ltd.\n\nAccepted 24 October 2011.\nAvailable online 04 November 2011.\nWe thank members of the Lester laboratory\nfor helpful discussion. This work was supported by National Institutes\nof Health (NIH) grants (DA17279, DA12242, DA015663,\nDA03194, MH53631, GM48677, and DA030396). R.M.D. was\nsupported by a fellowship from the California Tobacco Related\nDisease Research Program (15FT-0030), an NIH National Research\nService Award (DA021492), and an NIH Pathway to Independence\nAward (DA030396).\n\nAccepted Version - nihms-336935.pdf
", "abstract": "Firing rates of dopamine (DA) neurons in substantia nigra pars compacta (SNc) and ventral tegmental area (VTA) control DA release in target structures such as striatum and prefrontal cortex. DA neuron firing in the soma and release probability at axon terminals are tightly regulated by cholinergic transmission and nicotinic acetylcholine receptors (nAChRs). To understand the role of \u03b16* nAChRs in DA transmission, we studied several strains of mice expressing differing levels of mutant, hypersensitive (leucine 9\u2032 to serine [L9\u2032S]) \u03b16 subunits. \u03b16 L9\u2032S mice harboring six or more copies of the hypersensitive \u03b16 gene exhibited spontaneous home-cage hyperactivity and novelty-induced locomotor activity, whereas mice with an equal number of WT and L9\u2032S \u03b16 genes had locomotor activity resembling that of control mice. \u03b16-dependent, nicotine-stimulated locomotor activation was also more robust in high-copy \u03b16 L9\u2032S mice versus low-copy mice. In wheel-running experiments, results were also bi-modal; high-copy \u03b16 L9\u2032S animals exhibited blunted total wheel rotations during each day of a 9-day experiment, but low-copy \u03b16 L9\u2032S mice ran normally on the wheel. Reduced wheel running in hyperactive strains of \u03b16 L9\u2032S mice was attributable to a reduction in both overall running time and velocity. ACh and nicotine-stimulated DA release from striatal synaptosomes in \u03b16 L9\u2032S mice was well-correlated with behavioral phenotypes, supporting the hypothesis that augmented DA release mediates the altered behavior of \u03b16 L9\u2032S mice. This study highlights the precise control that the nicotinic cholinergic system exerts on DA transmission and provides further insights into the mechanisms and consequences of enhanced DA release.", "date": "2012-01-03", "date_type": "published", "publication": "Neuroscience", "volume": "200", "publisher": "Elsevier", "pagerange": "31-41", "id_number": "CaltechAUTHORS:20120214-082915834", "issn": "0306-4522", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120214-082915834", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "DA12242" }, { "agency": "NIH", "grant_number": "DA015663" }, { "agency": "NIH", "grant_number": "DA03194" }, { "agency": "NIH", "grant_number": "MH53631" }, { "agency": "NIH", "grant_number": "GM48677" }, { "agency": "NIH", "grant_number": "DA030396" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "15FT-0030" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "DA021492" }, { "agency": "NIH", "grant_number": "DA030396" } ] }, "doi": "10.1016/j.neuroscience.2011.10.047", "pmcid": "PMC3249511", "primary_object": { "basename": "nihms-336935.pdf", "url": "https://authors.library.caltech.edu/records/rx81d-bgj35/files/nihms-336935.pdf" }, "resource_type": "article", "pub_year": "2012", "author_list": "Cohen, B. N.; Mackey, E. D. W.; et el." }, { "id": "https://authors.library.caltech.edu/records/znfk0-1s981", "eprint_id": 25394, "eprint_status": "archive", "datestamp": "2023-08-22 03:57:12", "lastmod": "2023-10-24 15:50:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "McIntosh-J-M", "name": { "family": "McIntosh", "given": "J. Michael" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Characterizing functional \u03b16\u03b22 nicotinic acetylcholine receptors in vitro: Mutant \u03b22 subunits improve membrane expression, and fluorescent proteins reveal responsive cells", "ispublished": "pub", "full_text_status": "public", "keywords": "Nicotinic acetylcholine receptor; \u03b16; \u03b22; Fluorescent protein; Neuro2a cell; Endoplasmic reticulum exit sites", "note": "\u00a9 2011 Elsevier Inc. \n\nReceived 8 April 2011. Accepted 9 May 2011. Available online 17 May 2011. \n\nThis work was supported by grants from the US National Institutes of Health (DA17279, NS11756, AG033954, DA19375, DA12242, MH53631, and GM48677); from Targacept Inc.; from the California Tobacco-Related Disease Research Program (TRDRP); and from Louis and Janet Fletcher. R.S. was supported by a postdoctoral fellowship from TRDRP (18FT-0066), and R.D. by an NIH National Research Service Award (DA021492) and an NIH Pathway to Independence Award (DA030396).\n\nAccepted Version - nihms-303292.pdf
", "abstract": "\u03b16* nicotinic acetylcholine receptors (nAChRs) are highly expressed in mesostriatal and nigrostriatal dopaminergic systems, and participate in motor control, reward, and learning and memory. In vitro functional expression of \u03b16* nAChRs is essential for full pharmacological characterization of these receptors and for drug screening, but has been challenging. We expressed eGFP-tagged-\u03b16 and \u03b22 nAChR subunits in Neuro-2a cells, leading to functional channels. Inward currents were elicited with 300 \u03bcM ACh in 26% (5/19) of cells with evenly expressed \u03b16-eGFP in cytoplasm and periphery. We dramatically increased chances of detecting functional \u03b16-eGFP\u03b22 nAChRs by (i) introducing two endoplasmic reticulum (ER) export-enhancing mutations into \u03b22 subunits, and (ii) choosing cells with abundant Sec24D-mCherry-labeled ER exit sites. Both manipulations also modestly increased \u03b16-eGFP\u03b22 nAChR current amplitude. \u03b16-eGFP\u03b22 nAChRs were also activated by nicotine and by TC-2403. The \u03b16-eGFP\u03b22 currents were desensitized by 1 \u03bcM nicotine, blocked by \u03b1-conotoxin MII, partially inhibited by dihydro-\u03b2-erythroidine, and potentiated by extracellular Ca^(2+). Single-channel recordings showed that \u03b16-eGFP\u03b22 nAChRs had similar single-channel conductance to, but longer open time than, \u03b14-eGFP\u03b22 nAChRs. These methods provide avenues for developing cell lines expressing subtypes of \u03b16* nAChRs for both pharmacological study and drug screening.", "date": "2011-10-15", "date_type": "published", "publication": "Biochemical Pharmacology", "volume": "82", "number": "8", "publisher": "Elsevier", "pagerange": "852-861", "id_number": "CaltechAUTHORS:20110922-073823540", "issn": "0006-2952", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20110922-073823540", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "DA19375" }, { "agency": "NIH", "grant_number": "DA12242" }, { "agency": "NIH", "grant_number": "MH53631" }, { "agency": "NIH", "grant_number": "GM48677" }, { "agency": "Targacept Inc." }, { "agency": "Louis and Janet Fletcher" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "18FT-0066" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "DA021492" }, { "agency": "NIH", "grant_number": "DA030396" } ] }, "doi": "10.1016/j.bcp.2011.05.005", "pmcid": "PMC3162078", "primary_object": { "basename": "nihms-303292.pdf", "url": "https://authors.library.caltech.edu/records/znfk0-1s981/files/nihms-303292.pdf" }, "resource_type": "article", "pub_year": "2011", "author_list": "Xiao, Cheng; Srinivasan, Rahul; et el." }, { "id": "https://authors.library.caltech.edu/records/z6b55-75q60", "eprint_id": 27747, "eprint_status": "archive", "datestamp": "2023-09-18 23:03:16", "lastmod": "2023-10-23 22:39:37", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Du-Jiangang", "name": { "family": "Du", "given": "Jiangang" } }, { "id": "Blanche-T-J", "name": { "family": "Blanche", "given": "Timothy J." } }, { "id": "Harrison-R-R", "name": { "family": "Harrison", "given": "Reid R." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Masmanidis-S-C", "name": { "family": "Masmanidis", "given": "Sotiris C." } } ] }, "title": "Multiplexed, High Density Electrophysiology with Nanofabricated Neural Probes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2011 Du et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted\nuse, distribution, and reproduction in any medium, provided the original author and source are credited.\nReceived August 13, 2011; Accepted September 22, 2011; Published October 12, 2011.\nEditor: Hiromu Tanimoto, Max-Planck-Institut f\u00fcr Neurobiologie, Germany.\nFunding: J.D. and S.C.M. gratefully acknowledge support from the Broad Fellowship Program in Brain Circuitry and the Della Martin Fund for Discoveries in\nMental Illness. T.J.B. was supported by NIH-5R21NS066260. H.A.L. acknowledges funding by NIH DA017279. The funders had no role in study design, data\ncollection and analysis, decision to publish, or preparation of the manuscript.\n\nWe thank K. M. Scott and O. Mazor for advice on spike sorting, G.\nDeRose for assistance with electron-beam lithography, M. L. Roukes for\ndevice fabrication materials and equipment support, C. Xiao and A. D.\nSteele for animal support, and R. E. Penton for proofreading the\nmanuscript. We thank the staff at the Kavli Nanoscience Institute at\nCaltech, and the Nanoelectronics Research Facility at UCLA for device\nfabrication support.\nAuthor Contributions:\nConceived and designed the experiments: JD TJB RRH SCM. Performed\nthe experiments: JD TJB RRH SCM. Analyzed the data: JD TJB RRH\nSCM. Contributed reagents/materials/analysis tools: JD TJB RRH HAL\nSCM. Wrote the paper: JD TJB RRH HAL SCM.\n\nPublished - Du2011p16224Plos_One.pdf
Supplemental Material - FigureS1.tif
Supplemental Material - TableS1.docx
", "abstract": "Extracellular electrode arrays can reveal the neuronal network correlates of behavior with single-cell, single-spike, and sub-millisecond resolution. However, implantable electrodes are inherently invasive, and efforts to scale up the number and density of recording sites must compromise on device size in order to connect the electrodes. Here, we report on silicon-based neural probes employing nanofabricated, high-density electrical leads. Furthermore, we address the challenge of reading out multichannel data with an application-specific integrated circuit (ASIC) performing signal amplification, band-pass filtering, and multiplexing functions. We demonstrate high spatial resolution extracellular measurements with a fully integrated, low noise 64-channel system weighing just 330 mg. The on-chip multiplexers make possible recordings with substantially fewer external wires than the number of input channels. By combining nanofabricated probes with ASICs we have implemented a system for performing large-scale, high-density electrophysiology in small, freely behaving animals that is both minimally invasive and highly scalable.", "date": "2011-10-12", "date_type": "published", "publication": "PLoS ONE", "volume": "6", "number": "10", "publisher": "Public Library of Science", "pagerange": "Art. No. e26204", "id_number": "CaltechAUTHORS:20111111-102216534", "issn": "1932-6203", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20111111-102216534", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Broad Fellowship Program in Brain Circuitry" }, { "agency": "Della Martin Fund for Discoveries in Mental Illness" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "5R21NS066260" } ] }, "local_group": { "items": [ { "id": "Kavli-Nanoscience-Institute" } ] }, "doi": "10.1371/journal.pone.0026204", "pmcid": "PMC3192171", "primary_object": { "basename": "Du2011p16224Plos_One.pdf", "url": "https://authors.library.caltech.edu/records/z6b55-75q60/files/Du2011p16224Plos_One.pdf" }, "related_objects": [ { "basename": "FigureS1.tif", "url": "https://authors.library.caltech.edu/records/z6b55-75q60/files/FigureS1.tif" }, { "basename": "TableS1.docx", "url": "https://authors.library.caltech.edu/records/z6b55-75q60/files/TableS1.docx" } ], "resource_type": "article", "pub_year": "2011", "author_list": "Du, Jiangang; Blanche, Timothy J.; et el." }, { "id": "https://authors.library.caltech.edu/records/q7gs9-tt863", "eprint_id": 27277, "eprint_status": "archive", "datestamp": "2023-08-22 03:46:57", "lastmod": "2023-10-24 17:01:29", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Xu-J", "name": { "family": "Xu", "given": "J." } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "B. N." } }, { "id": "Zhu-Y", "name": { "family": "Zhu", "given": "Y." } }, { "id": "Dziewczapolski-G", "name": { "family": "Dziewczapolski", "given": "G." } }, { "id": "Panda-S", "name": { "family": "Panda", "given": "S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Heinemann-S-F", "name": { "family": "Heinemann", "given": "S. F." } }, { "id": "Contractor-A", "name": { "family": "Contractor", "given": "A." } } ] }, "title": "Altered activity\u2013rest patterns in mice with a human autosomal-dominant nocturnal frontal lobe epilepsy mutation in the \u03b22 nicotinic receptor", "ispublished": "pub", "full_text_status": "public", "keywords": "\u03b22^* nicotinic receptor; ADNFLE; knock-in mouse", "note": "\u00a9 2011 Macmillan Publishers Limited. \n\nReceived 18 September 2009; revised 13 April 2010; accepted 4. June 2010; published online 6 July 2010. \n\nThis work was supported by grants from NIH/NIDA (R01DA018247 to SFH, 5R01DA017279 to HAL), NIH/NEI (EY016807 to SP), NIH/NINDS (R01NS058894 to AC), NIH training awards (5T32NS041234-09 to JX, and 5T32EY007128-13 to YZ), and NS-011756 and NS-046464 to HAL. We thank Drs Michael Marks, Sharon Grady and Allan Collins for their suggestions.\n\nAccepted Version - nihms211620.pdf
", "abstract": "High-affinity nicotinic receptors containing \u03b22 subunits (\u03b22^*) are widely expressed in the brain, modulating many neuronal processes and contributing to neuropathologies such as Alzheimer's disease, Parkinson's disease and epilepsy. Mutations in both the \u03b14 and \u03b22 subunits are associated with a rare partial epilepsy, autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE). In this study, we introduced one such human missense mutation into the mouse genome to generate a knock-in strain carrying a valine-to-leucine mutation \u03b22V287L. \u03b22^(V287L) mice were viable and born at an expected Mendelian ratio. Surprisingly, mice did not show an overt seizure phenotype; however, homozygous mice did show significant alterations in their activity\u2013rest patterns. This was manifest as an increase in activity during the light cycle suggestive of disturbances in the normal sleep patterns of mice; a parallel phenotype to that found in human ADNFLE patients. Consistent with the role of nicotinic receptors in reward pathways, we found that \u03b22^(V287L) mice did not develop a normal proclivity to voluntary wheel running, a model for natural reward. Anxiety-related behaviors were also affected by the V287L mutation. Mutant mice spent more time in the open arms on the elevated plus maze suggesting that they had reduced levels of anxiety. Together, these findings emphasize several important roles of \u03b22^* nicotinic receptors in complex biological processes including the activity\u2013rest cycle, natural reward and anxiety.", "date": "2011-10", "date_type": "published", "publication": "Molecular Psychiatry", "volume": "16", "number": "10", "publisher": "Nature Publishing Group", "pagerange": "1048-1061", "id_number": "CaltechAUTHORS:20111018-104112346", "issn": "1359-4184", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20111018-104112346", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "R01DA018247" }, { "agency": "NIH", "grant_number": "5R01DA017279" }, { "agency": "NIH", "grant_number": "EY016807" }, { "agency": "NIH", "grant_number": "R01NS058894" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5T32NS041234-09" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5T32EY007128-13" }, { "agency": "NIH", "grant_number": "NS-011756" }, { "agency": "NIH", "grant_number": "NS-046464" } ] }, "doi": "10.1038/mp.2010.78", "pmcid": "PMC2970689", "primary_object": { "basename": "nihms211620.pdf", "url": "https://authors.library.caltech.edu/records/q7gs9-tt863/files/nihms211620.pdf" }, "resource_type": "article", "pub_year": "2011", "author_list": "Xu, J.; Cohen, B. N.; et el." }, { "id": "https://authors.library.caltech.edu/records/0xpq0-ggm32", "eprint_id": 27806, "eprint_status": "archive", "datestamp": "2023-08-19 08:13:45", "lastmod": "2023-10-24 17:26:28", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Van-Arman-E-B", "name": { "family": "Van Arnam", "given": "Ethan B." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Dissecting the Functions of Conserved Prolines within Transmembrane Helices of the D2 Dopamine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2011 American Chemical Society. \n\nReceived: May 13, 2011. Accepted: July 21, 2011. Publication Date (Web): July 21, 2011. \n\nThis work was supported by National Institutes of Health grant GM081662.\n\nAccepted Version - nihms-314331.pdf
Supplemental Material - cb200153g_si_001.pdf
", "abstract": "G protein-coupled receptors (GPCRs) contain a number of conserved proline residues in their transmembrane helices, and it is generally assumed these play important functional and/or structural roles. Here we use unnatural amino acid mutagenesis, employing \u03b1-hydroxy acids and proline analogues, to examine the functional roles of five proline residues in the transmembrane helices of the D2 dopamine receptor. The well-known tendency of proline to disrupt helical structure is important at all sites, while we find no evidence for a functional role for backbone amide cis\u2013trans isomerization, another feature associated with proline. At most proline sites, the loss of the backbone NH is sufficient to explain the role of the proline. However, at one site, P210^(5.50), a substituent on the backbone N appears to be essential for proper function. Interestingly, the pattern in functional consequences that we see is mirrored in the pattern of structural distortions seen in recent GPCR crystal structures.", "date": "2011-10", "date_type": "published", "publication": "ACS Chemical Biology", "volume": "6", "number": "10", "publisher": "American Chemical Society", "pagerange": "1063-1068", "id_number": "CaltechAUTHORS:20111116-105837023", "issn": "1554-8929", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20111116-105837023", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM081662" } ] }, "doi": "10.1021/cb200153g", "pmcid": "PMC3199346", "primary_object": { "basename": "cb200153g_si_001.pdf", "url": "https://authors.library.caltech.edu/records/0xpq0-ggm32/files/cb200153g_si_001.pdf" }, "related_objects": [ { "basename": "nihms-314331.pdf", "url": "https://authors.library.caltech.edu/records/0xpq0-ggm32/files/nihms-314331.pdf" } ], "resource_type": "article", "pub_year": "2011", "author_list": "Van Arnam, Ethan B.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/hvhqg-x6b21", "eprint_id": 27494, "eprint_status": "archive", "datestamp": "2023-08-19 08:12:34", "lastmod": "2023-10-24 17:11:51", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "S. C. R." }, "orcid": "0000-0001-9410-9805" }, { "id": "Thompson-A-J", "name": { "family": "Thompson", "given": "A. J." } }, { "id": "Bencherif-M", "name": { "family": "Bencherif", "given": "M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Varenicline Is a Potent Agonist of the Human 5-Hydroxytryptamine_3 Receptor", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2011 by the American Society for Pharmacology and Experimental Therapeutics. \n\nReceived June 21, 2011; accepted July 11, 2011. Published online before print July 20, 2011. \n\nThis work was supported by the Wellcome Trust [Grant 81925] (to A.J.T. and S.C.R.L.); the National Institutes of Health National Institute of Neurological Disorders and Stroke [Grant NS11756] (to H.A.L.); and the National Institutes of Health National Institute of General Medical Sciences [Grant GM19375] (to H.A.L.). S.C.R.L. is a Wellcome Trust Senior Research Fellow in Basic Biomedical Science. \nAuthorship Contributions: Participated in research design: Lummis, Thompson, Bencherif, and Lester. Conducted experiments: Lummis and Thompson. Performed data analysis: Lummis, Thompson, and Lester. Wrote or contributed to the writing of the manuscript: Lummis, Thompson, Bencherif, and Lester.", "abstract": "Varenicline, a widely used and successful smoking cessation agent, acts as a partial agonist at nicotinic acetylcholine receptors. Here, we explore the effects of varenicline at human and mouse 5-Hydroxytryptamine_3 (5-HT_3) receptors. Application of varenicline to human 5-HT_3 receptors expressed in Xenopus laevis oocytes reveal it is almost a full agonist (R_max = 80%) with an EC_50 (5.9 \u03bcM) 3-fold higher than 5-HT. At mouse 5-HT_3 receptors varenicline is a partial agonist (R_max = 35%) with an EC50 (18 \u03bcM) 20-fold higher than 5-HT. Displacement of the competitive 5-HT_3 receptor antagonist [^(3)H]granisetron reveals similar IC_50 values for varenicline at mouse and human receptors expressed in human embryonic kidney 293 cells, although studies in these cells using a membrane potential-sensitive dye show that again varenicline is a 4- or 35-fold less potent agonist than 5-HT in human and mouse receptors, respectively. Thus the data suggest that the efficacy, but not the affinity, of varenicline is greater at human 5-HT3 receptors compared with mouse. Docking studies provide a possible explanation for this difference, because they suggest distinct orientations of the ligand in the mouse versus human 5-HT_3 agonist binding sites. Additional binding selectivity studies in a broad panel of recombinant receptors and enzymes confirmed an interaction with 5-HT_3 receptors but revealed no additional interactions of varenicline. Therefore, activation of human 5-HT_3 receptors may be responsible for some of the side effects that preclude use of higher doses during varenicline treatment.", "date": "2011-10", "date_type": "published", "publication": "Journal of Pharmacology and Experimental Therapeutics", "volume": "339", "number": "1", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "125-131", "id_number": "CaltechAUTHORS:20111028-122557103", "issn": "0022-3565", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20111028-122557103", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust", "grant_number": "81925" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "GM19375" } ] }, "doi": "10.1124/jpet.111.185306", "pmcid": "PMC3186289", "resource_type": "article", "pub_year": "2011", "author_list": "Lummis, S. C. R.; Thompson, A. J.; et el." }, { "id": "https://authors.library.caltech.edu/records/zvw6s-prv20", "eprint_id": 25637, "eprint_status": "archive", "datestamp": "2023-08-22 03:42:43", "lastmod": "2023-10-24 15:58:09", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Blum-A-P", "name": { "family": "Blum", "given": "Angela P." } }, { "id": "Gleitsman-K-R", "name": { "family": "Gleitsman", "given": "Kristin Rule" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Evidence for an Extended Hydrogen Bond Network in the Binding Site of the Nicotinic Receptor: Role of the Vicinal Disulfide of the \u03b11 Subunit", "ispublished": "pub", "full_text_status": "public", "keywords": "Ion channels; Mutant; Nicotinic Acetylcholine Receptors; Protein Chemistry; Protein-Drug Interactions", "note": "\u00a9 2011 American Society for Biochemistry and Molecular Biology, Inc.\n\nReceived for publication, April 25, 2011, and in revised form, July 8, 2011 Published, JBC Papers in Press, July 13, 2011.\n\nThis work was supported, in whole or in part, by National Institutes of Health Grants NS 34407 (to D. A. D.) and NS 11756 (to H. A. L.).\n\nPublished - Blum2011p15944J_Biol_Chem.pdf
Supplemental Material - jbc.M111.254235-1.doc
", "abstract": "The defining feature of the \u03b1 subunits of the family of nicotinic acetylcholine receptors is a vicinal disulfide between Cys-192 and Cys-193. Although this structure has played a pivotal role in a number of pioneering studies of nicotinic receptors, its functional role in native receptors remains uncertain. Using mutant cycle analysis and unnatural residue mutagenesis, including backbone mutagenesis of the peptide bond of the vicinal disulfide, we have established the presence of a network of hydrogen bonds that extends from that peptide NH, across a \u03b2 turn to another backbone hydrogen bond, and then across the subunit interface to the side chain of a functionally important Asp residue in the non-\u03b1 subunit. We propose that the role of the vicinal disulfide is to distort the \u03b2 turn and thereby properly position a backbone NH for intersubunit hydrogen bonding to the key Asp.", "date": "2011-09-16", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "286", "number": "37", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "32251-32258", "id_number": "CaltechAUTHORS:20111004-120511603", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20111004-120511603", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1074/jbc.M111.254235", "pmcid": "PMC3173184", "primary_object": { "basename": "Blum2011p15944J_Biol_Chem.pdf", "url": "https://authors.library.caltech.edu/records/zvw6s-prv20/files/Blum2011p15944J_Biol_Chem.pdf" }, "related_objects": [ { "basename": "jbc.M111.254235-1.doc", "url": "https://authors.library.caltech.edu/records/zvw6s-prv20/files/jbc.M111.254235-1.doc" } ], "resource_type": "article", "pub_year": "2011", "author_list": "Blum, Angela P.; Gleitsman, Kristin Rule; et el." }, { "id": "https://authors.library.caltech.edu/records/fdpa8-72886", "eprint_id": 25403, "eprint_status": "archive", "datestamp": "2023-08-19 08:01:31", "lastmod": "2023-10-24 15:51:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Richards-C-I", "name": { "family": "Richards", "given": "Christopher I." } }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "Miwa-Julie-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Trafficking of \u03b14^* Nicotinic Receptors Revealed by Superecliptic Phluorin", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2011 by The American Society for Biochemistry and Molecular Biology, Inc. \n\nReceived for publication, April 29, 2011, and in revised form, July 13, 2011. \n\nThis work was supported, in whole or in part, by National Institutes of Health Grants AG033954, DA17279, and NS11756. This work was also supported by the California Tobacco-related Disease Research Program Grant 19KT-0032 and by a gift from Louis and Janet Fletcher. Supported by National Institutes of Health Kirschstein NRSA DA030877 and a Beckman Institute fellowship. Supported by a Tobacco-related Disease Research Program Postdoctoral Fellowship 18FT-0066 and by a Rapid Response grant from the Michael J. Fox Foundation.\n\nPublished - Richards2011p15814J_Biol_Chem.pdf
Supplemental Material - jbc.M111.256024-1.pdf
", "abstract": "We employed a pH-sensitive GFP analog, superecliptic phluorin, to observe aspects of nicotinic acetylcholine receptor (nAChR) trafficking to the plasma membrane (PM) in cultured mouse cortical neurons. The experiments exploit differences in the pH among endoplasmic reticulum (ER), trafficking vesicles, and the extracellular solution. The data confirm that few \u03b14\u03b24 nAChRs, but many \u03b14\u03b22 nAChRs, remain in neutral intracellular compartments, mostly the ER. We observed fusion events between nAChR-containing vesicles and PM; these could be quantified in the dendritic processes. We also studied the \u03b24R348C polymorphism, linked to amyotrophic lateral sclerosis (ALS). This mutation depressed fusion rates of \u03b14\u03b24 receptor-containing vesicles with the PM by \u223c2-fold, with only a small decrease in the number of nAChRs per vesicle. The mutation also decreased the number of ER exit sites, showing that the reduced receptor insertion results from a change at an early stage in trafficking. We confirm the previous report that the mutation leads to reduced agonist-induced currents; in the cortical neurons studied, the reduction amounts to 2\u20133-fold. Therefore, the reduced agonist-induced currents are caused by the reduced number of \u03b14\u03b24-containing vesicles reaching the membrane. Chronic nicotine exposure (0.2 \u03bcm) did not alter the PM insertion frequency or trafficking behavior of \u03b14\u03b24-laden vesicles. In contrast, chronic nicotine substantially increased the number of \u03b14\u03b22-containing vesicle fusions at the PM; this stage in \u03b14\u03b22 nAChR up-regulation is presumably downstream from increased ER exit. Superecliptic phluorin provides a tool to monitor trafficking dynamics of nAChRs in disease and addiction.", "date": "2011-09-09", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "286", "number": "36", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "31241-31249", "id_number": "CaltechAUTHORS:20110922-112958690", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20110922-112958690", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19KT-0032" }, { "agency": "Louis and Janet Fletcher" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "DA030877" }, { "agency": "Caltech Beckman Institute" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "18FT-0066" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "Arnold and Mabel Beckman Foundation" } ] }, "doi": "10.1074/jbc.M111.256024", "pmcid": "PMC3173132", "primary_object": { "basename": "Richards2011p15814J_Biol_Chem.pdf", "url": "https://authors.library.caltech.edu/records/fdpa8-72886/files/Richards2011p15814J_Biol_Chem.pdf" }, "related_objects": [ { "basename": "jbc.M111.256024-1.pdf", "url": "https://authors.library.caltech.edu/records/fdpa8-72886/files/jbc.M111.256024-1.pdf" } ], "resource_type": "article", "pub_year": "2011", "author_list": "Richards, Christopher I.; Srinivasan, Rahul; et el." }, { "id": "https://authors.library.caltech.edu/records/qpjds-bwk32", "eprint_id": 23522, "eprint_status": "archive", "datestamp": "2023-08-22 02:34:10", "lastmod": "2023-10-23 19:39:45", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Puskar-N-L", "name": { "family": "Puskar", "given": "Nyssa L." } }, { "id": "Xiu-Xinan", "name": { "family": "Xiu", "given": "Xinan" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Two Neuronal Nicotinic Acetylcholine Receptors, \u03b14\u03b24 and \u03b17, Show Differential Agonist Binding Modes", "ispublished": "pub", "full_text_status": "public", "keywords": "Drug Design; Nicotinic Acetylcholine Receptors; Protein Chemical Modification; Protein Drug Interactions; Protein Structure; Cation-\u03c0; Interaction; Unnatural Amino Acid Mutagenesis", "note": "\u00a9 2011 American Society for Biochemistry and Molecular \nBiology.\n\nReceived for publication, November 24, 2010, and in revised form, January 25, 2011. Published, JBC Papers in Press, February 22, 2011.\n\nThis work was supported, in whole or in part, by National Institutes of Health Grants NS 34407 and NS 11756.\n\nPublished - Puskar2011p13671J_Biol_Chem.pdf
Supplemental Material - jbc.M110.206565-1.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are pentameric, neurotransmitter-gated ion channels responsible for rapid excitatory neurotransmission in the central and peripheral nervous systems, resulting in skeletal muscle tone and various cognitive effects in the brain. These complex proteins are activated by the endogenous neurotransmitter ACh as well as by nicotine and structurally related agonists. Activation and modulation of nAChRs has been implicated in the pathology of multiple neurological disorders, and as such, these proteins are established therapeutic targets. Here we use unnatural amino acid mutagenesis to examine the ligand binding mechanisms of two homologous neuronal nAChRs: the \u03b14\u03b24 and \u03b17 receptors. Despite sequence identity among the residues that form the core of the agonist-binding site, we find that the \u03b14\u03b24 and \u03b17 nAChRs employ different agonist-receptor binding interactions in this region. The \u03b14\u03b24 receptor utilizes a strong cation-\u03c0 interaction to a conserved tryptophan (TrpB) of the receptor for both ACh and nicotine, and nicotine participates in a strong hydrogen bond with a backbone carbonyl contributed by TrpB. Interestingly, we find that the \u03b17 receptor also employs a cation-\u03c0 interaction for ligand recognition, but the site has moved to a different aromatic amino acid of the agonist-binding site depending on the agonist. ACh participates in a cation-\u03c0 interaction with TyrA, whereas epibatidine participates in a cation-\u03c0 interaction with TyrC2.", "date": "2011-04-22", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "286", "number": "16", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "14618-14627", "id_number": "CaltechAUTHORS:20110502-112328466", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20110502-112328466", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1074/jbc.M110.206565", "pmcid": "PMC3077659", "primary_object": { "basename": "Puskar2011p13671J_Biol_Chem.pdf", "url": "https://authors.library.caltech.edu/records/qpjds-bwk32/files/Puskar2011p13671J_Biol_Chem.pdf" }, "related_objects": [ { "basename": "jbc.M110.206565-1.pdf", "url": "https://authors.library.caltech.edu/records/qpjds-bwk32/files/jbc.M110.206565-1.pdf" } ], "resource_type": "article", "pub_year": "2011", "author_list": "Puskar, Nyssa L.; Xiu, Xinan; et el." }, { "id": "https://authors.library.caltech.edu/records/spgpt-b2k49", "eprint_id": 23728, "eprint_status": "archive", "datestamp": "2023-08-19 06:12:56", "lastmod": "2023-10-23 19:53:34", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Miwa-J-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Freedman-R", "name": { "family": "Freedman", "given": "Robert" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Neural Systems Governed by Nicotinic Acetylcholine Receptors: Emerging Hypotheses", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2011 Elsevier B.V. Available online 13 April 2011. We thank William Proctor and Susan Moriguchi for help with Figure 2 and T.K. Hensch, T.N. Wiesel, and R.L. Parker for helpful discussions. We received support from AG-33954, DA-11729, MH-86386, NS-11756, and the California\nTobacco-Related Disease Research Program (17RT-0127, 19KT-0032). J.M.M. is founder and shareholder of Ophidion, Inc. She has applied for U.S. patents 10322359 and 20080221013, on the use of lynx for therapeutic\npurposes. R.F. has received U.S. patent 10322359 on the use of alpha7 nAChR sequence variants in schizophrenia diagnosis. H.A.L. has received U.S. patent 6753456 on mice with hypersenitive alpha4 nicotinic receptors.\n\nAccepted Version - nihms683618.pdf
", "abstract": "Cholinergic neurons and nicotinic acetylcholine receptors (nAChRs) in the brain participate in diverse functions: reward, learning and memory, mood, sensory processing, pain, and neuroprotection. Nicotinic systems also have well-known roles in drug abuse. Here, we review recent insights into nicotinic function, linking exogenous and endogenous manipulations of nAChRs to alterations in synapses, circuits, and behavior. We also discuss how these contemporary advances can motivate attempts to exploit nicotinic systems therapeutically in Parkinson's disease, cognitive decline, epilepsy, and schizophrenia.", "date": "2011-04-14", "date_type": "published", "publication": "Neuron", "volume": "70", "number": "1", "publisher": "Elsevier", "pagerange": "20-33", "id_number": "CaltechAUTHORS:20110519-095350891", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20110519-095350891", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "17RT-0127" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "19KT-0032" }, { "agency": "NIH", "grant_number": "AG-33954" }, { "agency": "NIH", "grant_number": "DA-11729" }, { "agency": "NIH", "grant_number": "MH-86386" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1016/j.neuron.2011.03.014", "pmcid": "PMC4418790", "primary_object": { "basename": "nihms683618.pdf", "url": "https://authors.library.caltech.edu/records/spgpt-b2k49/files/nihms683618.pdf" }, "resource_type": "article", "pub_year": "2011", "author_list": "Miwa, Julie M.; Freedman, Robert; et el." }, { "id": "https://authors.library.caltech.edu/records/ywj7e-tx497", "eprint_id": 23223, "eprint_status": "archive", "datestamp": "2023-08-19 05:59:43", "lastmod": "2023-10-23 18:04:45", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Pless-S-A", "name": { "family": "Pless", "given": "Stephan A." } }, { "id": "Hanek-A-P", "name": { "family": "Hanek", "given": "Ariele P." } }, { "id": "Price-K-L", "name": { "family": "Price", "given": "Kerry L." } }, { "id": "Lynch-J-W", "name": { "family": "Lynch", "given": "Joseph W." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "Sarah C. R." }, "orcid": "0000-0001-9410-9805" } ] }, "title": "A Cation-\u03c0 Interaction at a Phenylalanine Residue in the Glycine Receptor Binding Site Is Conserved for Different Agonists", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2011 The American Society for Pharmacology and Experimental Therapeutics. Received October 22, 2010; accepted January 4, 2011. Published online before print January 25, 2011. This work was supported by the National Institutes of Health National Institute of Neurological Disorders and Stroke [Grants NS11756, NS34407]; the Wellcome Trust [Grant RG81925]; a Wellcome Trust Senior Research Fellowship in Basic Biomedical Science (to S.C.R.L.); the European Union (Grant FP7NeuroCypres); the Australian Research Council; and the National Health and Medical Research Council of Australia. \n\nAuthorship Contributions:\nParticipated in research design: Pless, Lynch, Lester, Dougherty,\nand Lummis.\nConducted experiments: Pless, Hanek, Price, and Lummis.\nContributed new reagents or analytic tools: Dougherty.\nPerformed data analysis: Pless, Hanek, Price, and Lummis.\nWrote or contributed to the writing of the manuscript: Pless,\nHanek, Price, Lynch, Lester, Dougherty, and Lummis.\nOther: Lynch, Lester, Dougherty, and Lummis acquired funding.", "abstract": "Cation-\u03c0 interactions have been demonstrated to play a major role in agonist-binding in Cys-loop receptors. However, neither the aromatic amino acid contributing to this interaction nor its location is conserved among Cys-loop receptors. Likewise, it is not clear how many different agonists of a given receptor form a cation-\u03c0 interaction or, if they do, whether it is with the same aromatic amino acid as the major physiological agonist. We demonstrated previously that Phe159 in the glycine receptor (GlyR) \u03b11 subunit forms a strong cation-\u03c0 interaction with the principal agonist, glycine. In the current study, we investigated whether the lower efficacy agonists of the human GlyR \u03b2-alanine and taurine also form cation-\u03c0 interactions with Phe159. By incorporating a series of unnatural amino acids, we found cation-\u03c0 interactions between Phe159 and the amino groups of \u03b2-alanine and taurine. The strengths of these interactions were significantly weaker than for glycine. Modeling studies suggest that \u03b2-alanine and taurine are orientated subtly differently in the binding pocket, with their amino groups further from Phe159 than that of glycine. These data therefore show that similar agonists can have similar but not identical orientations and interactions in the binding pocket and provide a possible explanation for the lower potencies of \u03b2-alanine and taurine.", "date": "2011-04", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "79", "number": "4", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "742-748", "id_number": "CaltechAUTHORS:20110404-100247422", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20110404-100247422", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "Wellcome Trust", "grant_number": "RG81925" }, { "agency": "European Union", "grant_number": "FP7 NeuroCypres" }, { "agency": "Australian Research Council" }, { "agency": "National Health and Medical Research Council (Australia)" } ] }, "doi": "10.1124/mol.110.069583", "pmcid": "PMC3063724", "resource_type": "article", "pub_year": "2011", "author_list": "Pless, Stephan A.; Hanek, Ariele P.; et el." }, { "id": "https://authors.library.caltech.edu/records/n1n1p-qjg75", "eprint_id": 22068, "eprint_status": "archive", "datestamp": "2023-08-19 05:05:09", "lastmod": "2023-10-23 15:39:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Srinivasan-Rahul", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Pantoja-R", "name": { "family": "Pantoja", "given": "Rigo" } }, { "id": "Moss-F-J", "name": { "family": "Moss", "given": "Fraser J." }, "orcid": "0000-0002-8519-6991" }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "Son-Cagdas-D", "name": { "family": "Son", "given": "Cagdas D." } }, { "id": "Miwa-Julie-M", "name": { "family": "Miwa", "given": "Julie" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Nicotine up-regulates \u03b14\u03b22 nicotinic receptors and ER exit sites via stoichiometry-dependent chaperoning", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2010 Srinivasan et al. This article is distributed under the terms of an Attribution\u2013\nNoncommercial\u2013Share Alike\u2013No Mirror Sites license for the first six months after the publication\ndate (see http://www.rupress.org/terms). After six months it is available under a\nCreative Commons License (Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license,\nas described at http://creativecommons.org/licenses/by-nc-sa/3.0/).\n\nSubmitted: 8 September 2010;\nAccepted: 9 December 2010.\nPublished December 27, 2010.\nWe thank Chris Richards and Larry Wade for technical assistance,\nand K. Scott and A. Goldsipe for assistance with MATLAB coding.\nThis work is supported by grants from the National Institutes\nof Health (NS-11756 and AG-033954); Targacept Inc.; Louis and\nJanet Fletcher; the Michael J. Fox Foundation (to R. Srinivasan);\nand the California Tobacco-Related Disease Research Program\n(18FT-0066 to R. Srinivasan).\nEdward N. Pugh Jr. served as editor.\n\nPublished - Srinivasan2011p12533J_Gen_Physiol.pdf
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Supplemental Material - J_Gen_Physiol_2011_Jan_137_1__59-79,_Figures.ppt
", "abstract": "The up-regulation of \u03b14\u03b22* nicotinic acetylcholine receptors (nAChRs) by chronic nicotine is a cell-delimited process and may be necessary and sufficient for the initial events of nicotine dependence. Clinical literature documents an inverse relationship between a person's history of tobacco use and his or her susceptibility to Parkinson's disease; this may also result from up-regulation. This study visualizes and quantifies the subcellular mechanisms involved in nicotine-induced nAChR up-regulation by using transfected fluorescent protein (FP)-tagged \u03b14 nAChR subunits and an FP-tagged Sec24D endoplasmic reticulum (ER) exit site marker. Total internal reflection fluorescence microscopy shows that nicotine (0.1 \u00b5M for 48 h) up-regulates \u03b14\u03b22 nAChRs at the plasma membrane (PM), despite increasing the fraction of \u03b14\u03b22 nAChRs that remain in near-PM ER. Pixel-resolved normalized F\u00f6rster resonance energy transfer microscopy between \u03b14-FP subunits shows that nicotine stabilizes the (\u03b14)_2(\u03b22)_3 stoichiometry before the nAChRs reach the trans-Golgi apparatus. Nicotine also induces the formation of additional ER exit sites (ERES). To aid in the mechanistic analysis of these phenomena, we generated a \u03b22_(enhanced-ER-export) mutant subunit that mimics two regions of the \u03b24 subunit sequence: the presence of an ER export motif and the absence of an ER retention/retrieval motif. The \u03b14\u03b22_(enhanced-ER-export) nAChR resembles nicotine-exposed nAChRs with regard to stoichiometry, intracellular mobility, ERES enhancement, and PM localization. Nicotine produces only small additional PM up-regulation of \u03b14\u03b22_(enhanced-ER-export) receptors. The experimental data are simulated with a model incorporating two mechanisms: (1) nicotine acts as a stabilizing pharmacological chaperone for nascent \u03b14\u03b22 nAChRs in the ER, eventually increasing PM receptors despite a bottleneck(s) in ER export; and (2) removal of the bottleneck (e.g., by expression of the \u03b22_(enhanced-ER-export) subunit) is sufficient to increase PM nAChR numbers, even without nicotine. The data also suggest that pharmacological chaperoning of nAChRs by nicotine can alter the physiology of ER processes.", "date": "2011-01", "date_type": "published", "publication": "Journal of General Physiology", "volume": "137", "number": "1", "publisher": "Rockefeller University Press", "pagerange": "59-79", "id_number": "CaltechAUTHORS:20110208-094115987", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20110208-094115987", "rights": "This article is distributed under the terms of an Attribution\u2013\nNoncommercial\u2013Share Alike\u2013No Mirror Sites license for the first six months after the publication\ndate (see http://www.rupress.org/terms). After six months it is available under a\nCreative Commons License (Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license,\nas described at http://creativecommons.org/licenses/by-nc-sa/3.0/).", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "AG-033954" }, { "agency": "Targacept Inc." }, { "agency": "Louis and Janet Fletcher" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "18FT-0066" } ] }, "doi": "10.1085/jgp.201010532", "pmcid": "PMC3010053", "primary_object": { "basename": "Srinivasan2011p12533J_Gen_Physiol.pdf", "url": "https://authors.library.caltech.edu/records/n1n1p-qjg75/files/Srinivasan2011p12533J_Gen_Physiol.pdf" }, "related_objects": [ { "basename": "JGP_201010532_V1.mov", "url": "https://authors.library.caltech.edu/records/n1n1p-qjg75/files/JGP_201010532_V1.mov" }, { "basename": "JGP_201010532_V2.mov", "url": "https://authors.library.caltech.edu/records/n1n1p-qjg75/files/JGP_201010532_V2.mov" }, { "basename": "JGP_201010532_V3.mov", "url": "https://authors.library.caltech.edu/records/n1n1p-qjg75/files/JGP_201010532_V3.mov" }, { "basename": "JGP_201010532_V4.mov", "url": "https://authors.library.caltech.edu/records/n1n1p-qjg75/files/JGP_201010532_V4.mov" }, { "basename": "JGP_201010532_V5.mov", "url": "https://authors.library.caltech.edu/records/n1n1p-qjg75/files/JGP_201010532_V5.mov" }, { "basename": "JGP_201010532_sm.pdf", "url": "https://authors.library.caltech.edu/records/n1n1p-qjg75/files/JGP_201010532_sm.pdf" }, { "basename": "J_Gen_Physiol_2011_Jan_137_1__59-79,_Figures.ppt", "url": "https://authors.library.caltech.edu/records/n1n1p-qjg75/files/J_Gen_Physiol_2011_Jan_137_1__59-79,_Figures.ppt" } ], "resource_type": "article", "pub_year": "2011", "author_list": "Srinivasan, Rahul; Pantoja, Rigo; et el." }, { "id": "https://authors.library.caltech.edu/records/1yr9e-cw445", "eprint_id": 21473, "eprint_status": "archive", "datestamp": "2023-08-19 04:14:41", "lastmod": "2023-10-21 00:04:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Thompson-A-J", "name": { "family": "Thompson", "given": "Andrew J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "Sarah C. R." }, "orcid": "0000-0001-9410-9805" } ] }, "title": "The structural basis of function in Cys-loop receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2010 Cambridge University Press.\nPublished online: 20 Sep 2010.\nThe authors' research is supported by the Wellcome Trust (SCRL, AJT; SCRL is a Wellcome\nTrust Senior Research Fellow in Basic Biomedical Science) ; the EU (SCRL; NeuroCypres FP7)\nand NIH (HAL).\n\nPublished - Thompson2010p12214Q_Rev_Biophys.pdf
", "abstract": "Cys-loop receptors are membrane-spanning neurotransmitter-gated ion channels that are responsible for fast excitatory and inhibitory transmission in the peripheral and central nervous systems. The best studied members of the Cys-loop family are nACh, 5-HT_3, GABA_A and glycine receptors. All these receptors share a common structure of five subunits, pseudo-symmetrically arranged to form a rosette with a central ion-conducting pore. Some are cation selective (e.g. nACh and 5-HT_3) and some are anion selective (e.g. GABA_A and glycine). Each receptor has an extracellular domain (ECD) that contains the ligand-binding sites, a transmembrane domain (TMD) that allows ions to pass across the membrane, and an intracellular domain (ICD) that plays a role in channel conductance and receptor modulation. Cys-loop receptors are the targets for many currently used clinically relevant drugs (e.g. benzodiazepines and anaesthetics). Understanding the molecular mechanisms of these receptors could therefore provide the catalyst for further development in this field, as well as promoting the development of experimental techniques for other areas of neuroscience.\nIn this review, we present our current understanding of Cys-loop receptor structure and function. The ECD has been extensively studied. Research in this area has been stimulated in recent years by the publication of high-resolution structures of nACh receptors and related proteins, which have permitted the creation of many Cys loop receptor homology models of this region. Here, using the 5-HT_3 receptor as a typical member of the family, we describe how homology modelling and ligand docking can provide useful but not definitive information about ligand interactions. We briefly consider some of the many Cys-loop receptors modulators. We discuss the current understanding of the structure of the TMD, and how this links to the ECD to allow channel gating, and consider the roles of the ICD, whose structure is poorly understood. We also describe some of the current methods that are beginning to reveal the differences between different receptor states, and may ultimately show structural details of transitions between them.", "date": "2010-11", "date_type": "published", "publication": "Quarterly Reviews of Biophysics", "volume": "43", "number": "4", "publisher": "Cambridge University Press", "pagerange": "449-499", "id_number": "CaltechAUTHORS:20101221-093346658", "issn": "0033-5835", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20101221-093346658", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "European Union (EU)", "grant_number": "NeuroCypres" }, { "agency": "NIH" } ] }, "doi": "10.1017/S0033583510000168", "primary_object": { "basename": "Thompson2010p12214Q_Rev_Biophys.pdf", "url": "https://authors.library.caltech.edu/records/1yr9e-cw445/files/Thompson2010p12214Q_Rev_Biophys.pdf" }, "resource_type": "article", "pub_year": "2010", "author_list": "Thompson, Andrew J.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/yvge7-ec843", "eprint_id": 19886, "eprint_status": "archive", "datestamp": "2023-08-19 03:18:48", "lastmod": "2023-10-20 22:00:55", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Blum-A-P", "name": { "family": "Blum", "given": "Angela P." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Nicotinic pharmacophore: The pyridine N of nicotine and carbonyl of acetylcholine hydrogen bond across a subunit interface to a backbone NH", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2010 by the National Academy of Sciences.\n\nThis contribution is part of the special series of Inaugural Articles by members of the National Academy of Sciences elected in 2009. \n\nContributed by Dennis A. Dougherty, June 2, 2010 (sent for review April 1, 2010).\n\nPublished online before print June 28, 2010.\n\nWe thank Ariele P. Hanek and Sean M. A. Kedrowski for\nhelpful discussions. This work was supported by the National Institutes of\nHealth (NS 34407; NS 11756) and the California Tobacco-Related Disease\nResearch Program of the University of California, Grant 16RT-0160.\nAuthor contributions: A.P.B. and D.A.D. designed research; A.P.B. performed research;\nA.P.B., H.A.L., and D.A.D. analyzed data; and A.P.B., H.A.L., and D.A.D. wrote the paper.\n\nPublished - Blum2010p11303P_Natl_Acad_Sci_Usa.pdf
Supplemental Material - pnas.1007140107_SI.pdf
", "abstract": "Pharmacophore models for nicotinic agonists have been proposed for four decades. Central to these models is the presence of a cationic nitrogen and a hydrogen bond acceptor. It is now well-established that the cationic center makes an important cation-\u03c0 interaction to a conserved tryptophan, but the donor to the proposed hydrogen bond acceptor has been more challenging to identify. A structure of nicotine bound to the acetylcholine binding protein predicted that the binding partner of the pharmacophore's second component was a water molecule, which also hydrogen bonds to the backbone of the complementary subunit of the receptors. Here we use unnatural amino acid mutagenesis coupled with agonist analogs to examine whether such a hydrogen bond is functionally significant in the \u03b14\u03b22 neuronal nAChR, the receptor most associated with nicotine addiction. We find evidence for the hydrogen bond with the agonists nicotine, acetylcholine, carbamylcholine, and epibatidine. These data represent a completed nicotinic pharmacophore and offer insight into the design of new therapeutic agents that selectively target these receptors.", "date": "2010-07-27", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "107", "number": "30", "publisher": "National Academy of Sciences", "pagerange": "13206-13211", "id_number": "CaltechAUTHORS:20100913-091409908", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20100913-091409908", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "California Tobacco-Related Disease Research Program of the University of California", "grant_number": "16RT-0160" } ] }, "doi": "10.1073/pnas.1007140107", "pmcid": "PMC2922130", "primary_object": { "basename": "Blum2010p11303P_Natl_Acad_Sci_Usa.pdf", "url": "https://authors.library.caltech.edu/records/yvge7-ec843/files/Blum2010p11303P_Natl_Acad_Sci_Usa.pdf" }, "related_objects": [ { "basename": "pnas.1007140107_SI.pdf", "url": "https://authors.library.caltech.edu/records/yvge7-ec843/files/pnas.1007140107_SI.pdf" } ], "resource_type": "article", "pub_year": "2010", "author_list": "Blum, Angela P.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/6kx5m-25s58", "eprint_id": 19388, "eprint_status": "archive", "datestamp": "2023-08-19 03:16:18", "lastmod": "2023-10-20 20:38:31", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Steele-A-D", "name": { "family": "Steele", "given": "Andrew D." } }, { "id": "McKinney-S", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Patzlaff-N-E", "name": { "family": "Patzlaff", "given": "Natalie E." } }, { "id": "McIntosh-J-M", "name": { "family": "McIntosh", "given": "J. Michael" } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Miwa-Julie-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Cholinergic Modulation of Locomotion and Striatal Dopamine Release Is Mediated by \u03b16\u03b14* Nicotinic Acetylcholine Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2010 the authors.\nReceived April 14, 2010; revised June 14, 2010; accepted June 16, 2010.\nThis work was supported by grants from the National Institutes of Health (NIH) (DA17279 and AG033954 to\nH.A.L.; DA19375 to H.A.L. and M.J.M.; DA12242, DA015663, and DA03194 to M.J.M.; MH53631 and GM48677 to J. M.\nMcIntosh), the Moore Foundation, the Croll Research Foundation (to J. M. Miwa), and the California Tobacco Related\nDisease Research Program (TRDRP; 12RT-0245 to H.A.L.). A.D.S. is funded by the Broad Fellow Program in Brain\nCircuitry at Caltech and an Ellison Medical Foundation New Scholar in Aging award. R.M.D. was supported by\npostdoctoral fellowships from TRDRP (15FT-0030) and NIH (DA021492 and NS007251). We thank members of the\nLester laboratory for helpful discussion. Thanks to P. Deshpande, M. Liu, C. Xiao, E. Mackey, G. Akopian, S. Benazouz,\nL. Sandoval, C. Baddick, and C. Wageman. We thank Oliver King and Cynthia Hsu for writing Matlab programs for\nhome cage behavior data analysis.\n\nPublished - Drenan2010p10955J_Neurosci.pdf
", "abstract": "Dopamine (DA) release in striatum is governed by firing rates of midbrain DA neurons, striatal cholinergic tone, and nicotinic ACh\nreceptors (nAChRs) on DA presynaptic terminals. DA neurons selectively express \u03b16* nAChRs, which show high ACh and nicotine\nsensitivity. To help identify nAChR subtypes that control DA transmission, we studied transgenic mice expressing hypersensitive \u03b16^(L9'S*)\nreceptors. \u03b16^(L9'S) mice are hyperactive, travel greater distance, exhibit increased ambulatory behaviors such as walking, turning, and\nrearing, and show decreased pausing, hanging, drinking, and grooming. These effects were mediated by \u03b16 \u03b14* pentamers, as \u03b16^(L9'S) mice\nlacking \u03b14 subunits displayed essentially normal behavior. In \u03b16^(L9'S) mice, receptor numbers are normal, but loss of \u03b14 subunits leads to\nfewer and less sensitive \u03b16* receptors. Gain-of-function nicotine-stimulated DA release from striatal synaptosomes requires \u03b14 subunits,\nimplicating \u03b16\u03b14\u03b22* nAChRs in \u03b16^(L9'S) mouse behaviors. In brain slices, we applied electrochemical measurements to study\ncontrol of DA release by \u03b16^(L9'S) nAChRs. Burst stimulation of DA fibers elicited increased DA release relative to single action potentials\nselectively in \u03b16^(L9'S), but not WT or \u03b14KO/ \u03b16^(L9'S), mice. Thus, increased nAChR activity, like decreased activity, leads to enhanced\nextracellular DA release during phasic firing. Bursts may directly enhance DA release from \u03b16^(L9'S) presynaptic terminals, as there was no\ndifference in striatal DA receptor numbers or DA transporter levels or function in vitro. These results implicate \u03b16\u03b14\u03b22* nAChRs in\ncholinergic control of DA transmission, and strongly suggest that these receptors are candidate drug targets for disorders involving the\nDA system.", "date": "2010-07-21", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "30", "number": "29", "publisher": "Society for Neuroscience", "pagerange": "9877-9889", "id_number": "CaltechAUTHORS:20100811-085614377", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20100811-085614377", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "DA19375" }, { "agency": "NIH", "grant_number": "DA12242" }, { "agency": "NIH", "grant_number": "DA015663" }, { "agency": "NIH", "grant_number": "DA03194" }, { "agency": "NIH", "grant_number": "MH53631" }, { "agency": "NIH", "grant_number": "GM48677" }, { "agency": "Moore Foundation" }, { "agency": "Croll Research Foundation" }, { "agency": "California Tobacco Related Disease Research Program (TRDRP)", "grant_number": "12RT-0245" }, { "agency": "California Tobacco Related Disease Research Program (TRDRP)", "grant_number": "15FT-0030" }, { "agency": "Caltech" }, { "agency": "Ellison Medical Foundation" }, { "agency": "NIH", "grant_number": "DA021492" }, { "agency": "NIH", "grant_number": "NS007251" } ] }, "doi": "10.1523/JNEUROSCI.2056-10.2010", "pmcid": "PMC3390922", "primary_object": { "basename": "Drenan2010p10955J_Neurosci.pdf", "url": "https://authors.library.caltech.edu/records/6kx5m-25s58/files/Drenan2010p10955J_Neurosci.pdf" }, "resource_type": "article", "pub_year": "2010", "author_list": "Drenan, Ryan M.; Grady, Sharon R.; et el." }, { "id": "https://authors.library.caltech.edu/records/npayz-87y77", "eprint_id": 18974, "eprint_status": "archive", "datestamp": "2023-08-19 02:55:52", "lastmod": "2023-10-20 19:15:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Hanek-A-P", "name": { "family": "Hanek", "given": "Ariele P." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Photochemical Proteolysis of an Unstructured Linker of the GABAAR Extracellular Domain Prevents GABA but Not Pentobarbital Activation", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2010 The American Society for Pharmacology and Experimental Therapeutics.\n\nReceived July 27, 2009; accepted April 2, 2010.\nPublished online before print April 2, 2010.\nWe thank Dr. Sarah Lummis for providing the human \u03b11 and \u03b22S\nGABAAR genes, for providing a GABAAR homology model, and for help in\nexperiment design. In addition, we thank Angela Blum for aid in the\nsynthesis of Npg and the preparation of the amino acid for coupling to dCA.\nThis work was supported by the National Institutes of Health National Institute of Neurological Disorders and Stroke [Grants NS34407, NS11756]. \n\nThis work was presented previously: Hanek AP (2009) Chemical-scale investigations\nof the Cys-loop neurotransmitter gated ion channel. Ph.D. thesis.\nCalifornia Institute of Technology, Pasadena, CA.", "abstract": "The GABA type A receptor (GABAAR) is the major inhibitory receptor in the mammalian central nervous system and the target of numerous pharmaceuticals. The \u03b1-subunit of these pentameric Cys-loop neurotransmitter-gated ion channels contributes to the binding of both GABA and allosteric modulators such as the benzodiazepines, suggesting a role for this subunit in the conformational changes associated with activation of the receptor. Herein we use the nonsense suppression methodology to incorporate a photoactivatable unnatural amino acid and photochemically cleave the backbone of the \u03b1 subunit of the \u03b1_1\u03b2_2 GABA_AR in a linker region that is believed to span the subunit. Proteolytic cleavage impairs GABA but not pentobarbital activation, strongly suggesting that conformational changes involving this linker region are critical to the GABA activation pathway.", "date": "2010-07", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "78", "number": "1", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "29-35", "id_number": "CaltechAUTHORS:20100709-141019159", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20100709-141019159", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH National Institute of Neurological Disorders and Stroke", "grant_number": "NS34407" }, { "agency": "NIH National Institute of Neurological Disorders and Stroke", "grant_number": "NS11756" } ] }, "doi": "10.1124/mol.109.059832", "pmcid": "PMC2912059", "resource_type": "article", "pub_year": "2010", "author_list": "Hanek, Ariele P.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/68ydd-wtv23", "eprint_id": 18567, "eprint_status": "archive", "datestamp": "2023-08-21 23:58:20", "lastmod": "2023-10-20 16:31:58", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Breining-S-R", "name": { "family": "Breining", "given": "Scott R." } }, { "id": "Yohannes-D", "name": { "family": "Yohannes", "given": "Daniel" } }, { "id": "Wageman-C-R", "name": { "family": "Wageman", "given": "Charles R." } }, { "id": "Fedorov-N-B", "name": { "family": "Fedorov", "given": "Nikolai B." } }, { "id": "McKinney-S", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Whiteaker-P", "name": { "family": "Whiteaker", "given": "Paul" } }, { "id": "Bencherif-M", "name": { "family": "Bencherif", "given": "Merouane" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } } ] }, "title": "Structural differences determine the relative selectivity of nicotinic compounds for native \u03b14\u03b22^*-, \u03b16\u03b22^*-, \u03b13\u03b24^*- and \u03b17-nicotine acetylcholine receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "TC2429; TC2403; TC1698; TC2242; TC6951; Varenicline", "note": "\u00a9 2010 Elsevier Ltd. \n\nReceived 2 July 2009. \nReceived 2 July 2009; revised 18 January 2010; accepted 21 January 2010. Available online 28 January 2010.\n\nPortions of this work were presented as an abstract at the Society for \nNicotine and Tobacco Research annual meeting 2008, rapid response \nposter: Synaptosomal assays as methods for identifying alpha6beta2*-\nnAChR selective compounds by SR Grady, C Wageman, I Fernandez, P \nWhiteaker, D Yohannes, M Bencherif, HA Lester, MJ Marks. \nThe authors thank Allan C. Collins for many helpful discussions. \nSupported by National Cooperative Drug Discovery Group U19 \nDA019375 from the National Institutes of Health to HAL, MJM and\nMB. The Institute for Behavioral Genetics animal colony is\nsupported by NIH grant DA015663 to Allan C. Collins and MJM.\nThree of the authors (SRB, DY, MB) are employees of Targacept, \nInc., which holds patents on several of the compounds studied in \nthis paper. All studies conducted at Targacept, Inc., and reported \nhere were supported by DA019375.\n\nAccepted Version - nihms175400.pdf
Supplemental Material - f.doc
Supplemental Material - f.ppt
", "abstract": "Mammalian brain expresses multiple nicotinic acetylcholine receptor (nAChR) subtypes that differ in subunit\ncomposition, sites of expression and pharmacological and functional properties. Among known subtypes of\nreceptors, \u03b14\u03b22^* and \u03b16\u03b22^*-nAChR have the highest affinity for nicotine (where ^* indicates possibility of other\nsubunits). The \u03b14\u03b22^*-nAChRs are widely distributed, while \u03b16\u03b22^*-nAChR are restricted to a few regions. Both\nsubtypes modulate release of dopamine from the dopaminergic neurons of the mesoaccumbens pathway\nthought to be essential for reward and addiction. \u03b14\u03b22^*-nAChR also modulate GABA release in these areas.\nIdentification of selective compounds would facilitate study of nAChR subtypes. An improved understanding\nof the role of nAChR subtypes may help in developing more effective smoking cessation aids with\nfewer side effects than current therapeutics.We have screened a series of nicotinic compounds that vary in\nthe distance between the pyridine and the cationic center, in steric bulk, and in flexibility of the molecule.\nThese compoundswere screened usingmembrane binding and synaptosomal function assays, or recordings\nfrom GH4C1 cells expressing h\u03b17, to determine affinity, potency and efficacy at four subtypes of nAChRs\nfound in brain, \u03b14\u03b22^*, \u03b16\u03b22^*, \u03b17 and \u03b13\u03b24^*. In addition, physiological assays in gain-of-function mutant\nmice were used to assess in vivo activity at \u03b14b2^* and \u03b16\u03b22^*-nAChRs. This approach has identified several\ncompounds with agonist or partial agonist activity that display improved selectivity for \u03b16\u03b22^*-nAChR.", "date": "2010-06", "date_type": "published", "publication": "Neuropharmacology", "volume": "58", "number": "7", "publisher": "Elsevier", "pagerange": "1054-1066", "id_number": "CaltechAUTHORS:20100604-114525877", "issn": "0028-3908", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20100604-114525877", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "U19 DA019375" }, { "agency": "NIH", "grant_number": "DA015663" }, { "agency": "NIH", "grant_number": "DA019375" } ] }, "doi": "10.1016/j.neuropharm.2010.01.013", "pmcid": "PMC2849849", "primary_object": { "basename": "f.doc", "url": "https://authors.library.caltech.edu/records/68ydd-wtv23/files/f.doc" }, "related_objects": [ { "basename": "f.ppt", "url": "https://authors.library.caltech.edu/records/68ydd-wtv23/files/f.ppt" }, { "basename": "nihms175400.pdf", "url": "https://authors.library.caltech.edu/records/68ydd-wtv23/files/nihms175400.pdf" } ], "resource_type": "article", "pub_year": "2010", "author_list": "Grady, Sharon R.; Drenan, Ryan M.; et el." }, { "id": "https://authors.library.caltech.edu/records/fr30f-zdz54", "eprint_id": 17916, "eprint_status": "archive", "datestamp": "2023-08-21 23:35:35", "lastmod": "2023-10-20 15:25:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Limapichat-W", "name": { "family": "Limapichat", "given": "Walrati" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Chemical Scale Studies of the Phe-Pro Conserved Motif in the Cys Loop of Cys", "ispublished": "pub", "full_text_status": "public", "keywords": "Peptides/Conformation; Receptors/7-Helix Ligand-gated Channels; Receptors/Structure-Function; Ion Channels; Nicotinic Acetylcholine Receptors; Cys Loop; Nicotinic Receptor; Proline; Unnatural Amino Acid Mutagenesis", "note": "\u00a9 2010 American Society for Biochemistry and Molecular Biology. \n\nReceived August 28, 2009; revision received January 8, 2010. \n\nThis work was supported, in whole or in part, by National Institutes of Health Grants NS-34407 and NS-11756. We thank Dr. Scott A. Ross for help with the NMR experiments and Professor Sarah C. R. Lummis for helpful discussion.\n\nPublished - Limapichat2010p7377Journal_of_Biological_Chemistry.pdf
Supplemental Material - jbc.M109.060939-1.doc
", "abstract": "The functions of two conserved residues, Phe^(135) and Pro^(136), located at the apex of the Cys loop of the nicotinic acetylcholine receptor are investigated. Both residues were substituted with natural and unnatural amino acids, focusing on the role of aromaticity at Phe^(135), backbone conformation at Pro^(136), side chain polarity and volume, and the specific interaction between the aromatic side chain and the proline. NMR spectroscopy studies of model peptides containing proline and unnatural proline analogues following a Phe show a consistent increase in the population of the cis conformer relative to peptides lacking the Phe. In the receptor, a strong interaction between the Phe and Pro residues is evident, as is a strong preference for aromaticity and hydrophobicity at the Phe site. A similar influence of hydrophobicity is observed at the proline site. In addition, across a simple homologous series of proline analogues, the results reveal a correlation between receptor function and cis bias at the proline backbone. This could suggest a significant role for the cis proline conformer at this site in receptor function.", "date": "2010-03-19", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "285", "number": "12", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "8976-8984", "id_number": "CaltechAUTHORS:20100409-120603576", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20100409-120603576", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1074/jbc.M109.060939", "pmcid": "PMC2838319", "primary_object": { "basename": "Limapichat2010p7377Journal_of_Biological_Chemistry.pdf", "url": "https://authors.library.caltech.edu/records/fr30f-zdz54/files/Limapichat2010p7377Journal_of_Biological_Chemistry.pdf" }, "related_objects": [ { "basename": "jbc.M109.060939-1.doc", "url": "https://authors.library.caltech.edu/records/fr30f-zdz54/files/jbc.M109.060939-1.doc" } ], "resource_type": "article", "pub_year": "2010", "author_list": "Limapichat, Walrati; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/hj2wr-xkr71", "eprint_id": 17209, "eprint_status": "archive", "datestamp": "2023-08-21 23:05:55", "lastmod": "2023-10-19 23:39:17", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Zhao-Shea-R", "name": { "family": "Zhao-Shea", "given": "Rubing" } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Just-H", "name": { "family": "Just", "given": "Herwig" } }, { "id": "McClure-Begley-T", "name": { "family": "McClure-Begley", "given": "Tristan" } }, { "id": "Whiteaker-P", "name": { "family": "Whiteaker", "given": "Paul" } }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Salminem-O", "name": { "family": "Salminem", "given": "Outi" } }, { "id": "Gardner-P-D", "name": { "family": "Gardner", "given": "Paul D." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Tapper-A-R", "name": { "family": "Tapper", "given": "Andrew R." } } ] }, "title": "Dopamine D_2-receptor activation elicits akinesia, rigidity, catalepsy, and tremor in mice expressing hypersensitive 4 nicotinic receptors via a cholinergic-dependent mechanism", "ispublished": "pub", "full_text_status": "restricted", "keywords": "acetylcholine; striatum; nicotine; parkinsonian; interneuron", "note": "\u00a9 2010 Federation of American Societies for Experimental Biology. \n\nReceived for publication May 26, 2009; accepted for publication August 6, 2009.", "abstract": "Recent studies suggest that high-affinity neuronal nicotinic acetylcholine receptors (nAChRs) containing \u03b14 and \u03b22 subunits (\u03b14\u03b22*) functionally interact with G-protein-coupled dopamine (DA) D_2 receptors in basal ganglia. We hypothesized that if a functional interaction between these receptors exists, then mice expressing an M2 point mutation (Leu9'Ala) rendering 4 nAChRs hypersensitive to ACh may exhibit altered sensitivity to a D_2-receptor agonist. When challenged with the D_(2)R agonist, quinpirole (0.5\u201310 mg/kg), Leu9'Ala mice, but not wild-type (WT) littermates, developed severe, reversible motor impairment characterized by rigidity, catalepsy, akinesia, and tremor. While striatal DA tissue content, baseline release, and quinpirole-induced DA depletion did not differ between Leu9'Ala and WT mice, quinpirole dramatically increased activity of cholinergic striatal interneurons only in mutant animals, as measured by increased c-Fos expression in choline acetyltransferase (ChAT)-positive interneurons. Highlighting the importance of the cholinergic system in this mouse model, inhibiting the effects of ACh by blocking muscarinic receptors, or by selectively activating hypersensitive nAChRs with nicotine, rescued motor symptoms. This novel mouse model mimics the imbalance between striatal DA/ACh function associated with severe motor impairment in disorders such as Parkinson's disease, and the data suggest that a D_(2)R\u2013\u03b14*-nAChR functional interaction regulates cholinergic interneuron activity.\u2014Zhao-Shea, R., Cohen, B. N., Just, H., McClure-Begley, T., Whiteaker, P., Grady, S. R., Salminen, O., Gardner, P. D., Lester, H. A., Tapper, A. R. Dopamine D2-receptor activation elicits akinesia, rigidity, catalepsy, and tremor in mice expressing hypersensitive \u03b14 nicotinic receptors via a cholinergic-dependent mechanism.", "date": "2010-01", "date_type": "published", "publication": "FASEB Journal", "volume": "24", "number": "1", "publisher": "Federation of American Societies for Experimental Biology", "pagerange": "49-57", "id_number": "CaltechAUTHORS:20100119-103312477", "issn": "0892-6638", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20100119-103312477", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Institute of Neurological Disorders and Stroke", "grant_number": "NS059586" }, { "agency": "National Institute on Drug Abuse", "grant_number": "DA017279" }, { "agency": "National Institute on Drug Abuse", "grant_number": "DA12242" }, { "agency": "National Institute on Aging", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "DA015663" } ] }, "doi": "10.1096/fj.09-137034", "pmcid": "PMC2797035", "resource_type": "article", "pub_year": "2010", "author_list": "Zhao-Shea, Rubing; Cohen, Bruce N.; et el." }, { "id": "https://authors.library.caltech.edu/records/7efv8-p9e38", "eprint_id": 17002, "eprint_status": "archive", "datestamp": "2023-08-19 00:38:21", "lastmod": "2023-10-19 22:43:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Moss-F-J", "name": { "family": "Moss", "given": "Fraser J." }, "orcid": "0000-0002-8519-6991" }, { "id": "Imoukhuede-P-I", "name": { "family": "Imoukhuede", "given": "P. I." } }, { "id": "Scott-Kimberly", "name": { "family": "Scott", "given": "Kimberly" } }, { "id": "Hu-Jia", "name": { "family": "Hu", "given": "Jia" } }, { "id": "Jankowsky-J-L", "name": { "family": "Jankowsky", "given": "Joanna L." } }, { "id": "Quick-M-W", "name": { "family": "Quick", "given": "Michael W." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "GABA transporter function, oligomerization state, and anchoring: correlates with subcellularly resolved FRET", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2009 Moss et al.\nThis article is distributed under the terms of an Attribution\u2013Noncommercial\u2013\nShare Alike\u2013No Mirror Sites license for the first six months after the publication\ndate (see http://www.jgp.org/misc/terms.shtml). After six months it is available under a Creative\nCommons License (Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as\ndescribed at http://creativecommons.org/licenses/by-nc-sa/3.0/).\n\nSubmitted: 7 August 2009.\nAccepted: 2 November 2009.\n\nEdward N. Pugh Jr. served as editor.\n\n\nWe thank Robert Chow, Cagdas Son, Rigo Pantoja, and Rahul\nSrinivasan for discussion. We thank Jo Ann Trinkle and Elisha\nMackey for technical assistance. We also acknowledge Ryan\nMorin of Michael Smith Genome Sciences Centre, BC Cancer\nAgency Vancouver, BC, Canada for his assistance with the\nbioinformatics.\nThis research is supported by grants from the National Institutes\nof Health (grants DA-09121, DA-10509, and NS-11756). F.J.\nMoss received an American Heart Association Postdoctoral Fellowship.\nSome experiments used the facilities of the Millard and\nMuriel Jacobs Genetics and Genomics Laboratory at the California\nInstitute of Technology.\n\nPublished - Moss2009p6559J_Gen_Physiol.pdf
Supplemental Material - Moss2009p6559J_Gen_Physiol_supp.pdf
", "abstract": "The mouse \u03b3-aminobutyric acid (GABA) transporter mGAT1 was expressed in neuroblastoma 2a cells. 19 mGAT1 designs incorporating fluorescent proteins were functionally characterized by [^3H]GABA uptake in assays that responded to several experimental variables, including the mutations and pharmacological manipulation of the cytoskeleton. Oligomerization and subsequent trafficking of mGAT1 were studied in several subcellular regions of live cells using localized fluorescence, acceptor photobleach F\u00f6rster resonance energy transfer (FRET), and pixel-by-pixel analysis of normalized FRET (NFRET) images. Nine constructs were functionally indistinguishable from wild-type mGAT1 and provided information about normal mGAT1 assembly and trafficking. The remainder had compromised [^3H]GABA uptake due to observable oligomerization and/or trafficking deficits; the data help to determine regions of mGAT1 sequence involved in these processes. Acceptor photobleach FRET detected mGAT1 oligomerization, but richer information was obtained from analyzing the distribution of all-pixel NFRET amplitudes. We also analyzed such distributions restricted to cellular subregions. Distributions were fit to either two or three Gaussian components. Two of the components, present for all mGAT1 constructs that oligomerized, may represent dimers and high-order oligomers (probably tetramers), respectively. Only wild-type functioning constructs displayed three components; the additional component apparently had the highest mean NFRET amplitude. Near the cell periphery, wild-type functioning constructs displayed the highest NFRET. In this subregion, the highest NFRET component represented ~30% of all pixels, similar to the percentage of mGAT1 from the acutely recycling pool resident in the plasma membrane in the basal state. Blocking the mGAT1 C terminus postsynaptic density 95/discs large/zona occludens 1 (PDZ)-interacting domain abolished the highest amplitude component from the NFRET distributions. Disrupting the actin cytoskeleton in cells expressing wild-type functioning transporters moved the highest amplitude component from the cell periphery to perinuclear regions. Thus, pixel-by-pixel NFRET analysis resolved three distinct forms of GAT1: dimers, high-order oligomers, and transporters associated via PDZ-mediated interactions with the actin cytoskeleton and/or with the exocyst.", "date": "2009-12", "date_type": "published", "publication": "Journal of General Physiology", "volume": "134", "number": "6", "publisher": "Rockefeller University Press", "pagerange": "489-521", "id_number": "CaltechAUTHORS:20091218-122259894", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20091218-122259894", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "DA-10509" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "American Heart Association" } ] }, "doi": "10.1085/jgp.200910314", "pmcid": "PMC2806419", "primary_object": { "basename": "Moss2009p6559J_Gen_Physiol.pdf", "url": "https://authors.library.caltech.edu/records/7efv8-p9e38/files/Moss2009p6559J_Gen_Physiol.pdf" }, "related_objects": [ { "basename": "Moss2009p6559J_Gen_Physiol_supp.pdf", "url": "https://authors.library.caltech.edu/records/7efv8-p9e38/files/Moss2009p6559J_Gen_Physiol_supp.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Moss, Fraser J.; Imoukhuede, P. I.; et el." }, { "id": "https://authors.library.caltech.edu/records/gexgz-fnn74", "eprint_id": 16535, "eprint_status": "archive", "datestamp": "2023-08-19 00:17:22", "lastmod": "2023-10-19 22:20:49", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Cai-Haijiang", "name": { "family": "Cai", "given": "Haijiang" } }, { "id": "McIntosh-J-M", "name": { "family": "McIntosh", "given": "J. Michael" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Chronic Nicotine Selectively Enhances \u03b14\u03b22* Nicotinic Acetylcholine Receptors in the Nigrostriatal Dopamine Pathway", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2009 Society for Neuroscience. \n\nReceived June 20, 2009; revised Aug. 22, 2009; accepted Aug. 29, 2009. \n\nThis work was supported by grants from the U.S. National Institutes of Health (DA17279, AG033954, MH53631), from Targacept Inc., and from Louis and Janet Fletcher. We also acknowledge support from the Natural Sciences and Engineering Research Council Canada (R.N.), the NARSAD Young Investigator Program (R.N.), and the California Tobacco-Related Disease Research Program (16FT-0066 to C.X.). We thank J. Drago for 4 knock-out mice, P. Deshpande and C. D. Son for much assistance, and R. Srinivasan and B. N. Cohen for comments.\n\nPublished - Xiao2009p6184J_Neurosci.pdf
Supplemental Material - Xiao2009p6184J_Neurosci_supp.pdf
Supplemental Material - Xiao2009p6184J_Neurosci_supp_2.pdf
", "abstract": "These electrophysiological experiments, in slices and intact animals, study the effects of in vivo chronic exposure to nicotine on functional \u03b14\u03b22* nAChRs in the nigrostriatal dopaminergic (DA) pathway. Recordings were made in wild-type and \u03b14 nicotinic acetylcholine receptor (nAChR) subunit knock-out mice. Chronic nicotine enhanced methyllycaconitine citrate hydrate-resistant, dihydro-\u03b2-erythroidine hydrobromide-sensitive nicotinic currents elicited by 3\u20131000 \u00b5M ACh in GABAergic neurons of the substantia nigra pars reticulata (SNr), but not in DA neurons of the substantia nigra pars compacta (SNc). This enhancement leads to higher firing rates of SNr GABAergic neurons and consequently to increased GABAergic inhibition of the SNc DA neurons. In the dorsal striatum, functional \u03b14* nAChRs were not found on the neuronal somata; however, nicotine acts via \u03b14\u03b22* nAChRs in the DA terminals to modulate glutamate release onto the medium spiny neurons. Chronic nicotine also increased the number and/or function of these \u03b14\u03b22* nAChRs. These data suggest that in nigrostriatal DA pathway, chronic nicotine enhancement of \u03b14\u03b22* nAChRs displays selectivity in cell type and in nAChR subtype as well as in cellular compartment. These selective events augment inhibition of SNc DA neurons by SNr GABAergic neurons and also temper the release of glutamate in the dorsal striatum. The effects may reduce the risk of excitotoxicity in SNc DA neurons and may also counteract the increased effectiveness of corticostriatal glutamatergic inputs during degeneration of the DA system. These processes may contribute to the inverse correlation between tobacco use and Parkinson's disease.", "date": "2009-10-07", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "29", "number": "40", "publisher": "Society for Neuroscience", "pagerange": "12428-12439", "id_number": "CaltechAUTHORS:20091102-100009481", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20091102-100009481", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "MH53631" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "National Alliance for Research on Schizophrenia & Depression" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "16FT-0066" } ] }, "doi": "10.1523/JNEUROSCI.2939-09.2009", "pmcid": "PMC2787412", "primary_object": { "basename": "Xiao2009p6184J_Neurosci_supp.pdf", "url": "https://authors.library.caltech.edu/records/gexgz-fnn74/files/Xiao2009p6184J_Neurosci_supp.pdf" }, "related_objects": [ { "basename": "Xiao2009p6184J_Neurosci_supp_2.pdf", "url": "https://authors.library.caltech.edu/records/gexgz-fnn74/files/Xiao2009p6184J_Neurosci_supp_2.pdf" }, { "basename": "Xiao2009p6184J_Neurosci.pdf", "url": "https://authors.library.caltech.edu/records/gexgz-fnn74/files/Xiao2009p6184J_Neurosci.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Xiao, Cheng; Nashmi, Raad; et el." }, { "id": "https://authors.library.caltech.edu/records/tw984-wb187", "eprint_id": 15865, "eprint_status": "archive", "datestamp": "2023-08-19 00:15:34", "lastmod": "2023-10-19 17:19:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Pantoja-R", "name": { "family": "Pantoja", "given": "Rigo" } }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Kadambi-S", "name": { "family": "Kadambi", "given": "Sindhuja" } }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "Tzlil-S", "name": { "family": "Tzlil", "given": "Shelly" } }, { "id": "Moss-F-J", "name": { "family": "Moss", "given": "Fraser J." }, "orcid": "0000-0002-8519-6991" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Pharmacological chaperoning of nicotinic receptors begins in the endoplasmic reticulum: High-resolution imaging", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2009 Elsevier. \n\nR.P. and R.S. contributed equally to this work. Grants: NS11756, DA17279, Michael J. Fox, Philip Morris, Targacept. Fellowships: Ford and APA-DPN (RP). AHA postdoctoral (FJM).", "abstract": "[No abstract]", "date": "2009-10-01", "date_type": "published", "publication": "Biochemical Pharmacology", "volume": "78", "number": "7", "publisher": "Elsevier", "pagerange": "900-901", "id_number": "CaltechAUTHORS:20090916-091123236", "issn": "0006-2952", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090916-091123236", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "Philip Morris" }, { "agency": "Targacept" }, { "agency": "Ford Foundation" }, { "agency": "American Psychological Association" }, { "agency": "American Heart Association" } ] }, "doi": "10.1016/j.bcp.2009.06.033", "resource_type": "article", "pub_year": "2009", "author_list": "Pantoja, Rigo; Srinivasan, Rahul; et el." }, { "id": "https://authors.library.caltech.edu/records/e4g1b-afr32", "eprint_id": 15820, "eprint_status": "archive", "datestamp": "2023-08-19 00:09:46", "lastmod": "2023-10-19 17:17:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Pantoja-R", "name": { "family": "Pantoja", "given": "Rigo" } }, { "id": "Kadambi-S", "name": { "family": "Kadambi", "given": "Sindhuja" } }, { "id": "Mackey-E-D-W", "name": { "family": "Mackey", "given": "Elisha D. W." } }, { "id": "Tzlil-S", "name": { "family": "Tzlil", "given": "Shelly" } }, { "id": "Moss-F-J", "name": { "family": "Moss", "given": "Fraser J." }, "orcid": "0000-0002-8519-6991" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Pharmacological chaperoning of nicotinic receptors begins in the endoplasmic reticulum: Compartments and stoichiometries", "ispublished": "pub", "full_text_status": "restricted", "note": "Copyright \u00a9 2009 Elsevier. \n\nAvailable online 13 August 2009.\n \nR.P. and R.S. contributed equally to this work. Grants: NS11756, DA17279, Michael J. Fox Foundation (RS), Philip Morris, Targacept. Fellowships: Ford and APA-DPN (RP). AHA postdoctoral (FJM).", "abstract": "[Poster abstract]", "date": "2009-10", "date_type": "published", "publication": "Biochemical Pharmacology", "volume": "78", "number": "7", "publisher": "Elsevier", "pagerange": "900", "id_number": "CaltechAUTHORS:20090911-153603948", "issn": "0006-2952", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090911-153603948", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "Philip Morris" }, { "agency": "Targacept" }, { "agency": "Ford Foundation" }, { "agency": "American Psychological Association" }, { "agency": "American Heart Association" } ] }, "doi": "10.1016/j.bcp.2009.06.032", "resource_type": "article", "pub_year": "2009", "author_list": "Srinivasan, Rahul; Pantoja, Rigo; et el." }, { "id": "https://authors.library.caltech.edu/records/6sfz9-yw476", "eprint_id": 14738, "eprint_status": "archive", "datestamp": "2023-08-21 21:59:45", "lastmod": "2023-10-18 18:45:36", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Breining-S-R", "name": { "family": "Breining", "given": "Scott R." } }, { "id": "Bencherif-M", "name": { "family": "Bencherif", "given": "Merouane" } }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Whiteaker-P", "name": { "family": "Whiteaker", "given": "Paul" } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Wageman-C-R", "name": { "family": "Wageman", "given": "Charles R." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Yohannes-D", "name": { "family": "Yohannes", "given": "Daniel" } } ] }, "title": "Evaluation of structurally diverse neuronal nicotinic receptor ligands for selectivity at the \u03b16 subtype", "ispublished": "pub", "full_text_status": "public", "keywords": "Neuronal nicotinic receptor; NNR; Alpha 6; a3b4; SAR; Pyrimidine; Nicotinic acetylcholine receptor; Smoking cessation; Nicotine addiction; Dopamine release; Parkinson's disease", "note": "\u00a9 2009 Elsevier Ltd. \n\nReceived 16 March 2009; revised 15 May 2009; accepted 20 May 2009. Available online 27 May 2009. \n\nThis work was supported by NCDDG grant DA019375 from the National Institutes of Health to H.A.L., M.J.M. and M.B.\n\nAccepted Version - nihms-128262.pdf
Supplemental Material - mmc1.doc
", "abstract": "Direct comparison of pyridine versus pyrimidine substituents on a small but diverse set of ligands indicates that the pyrimidine substitution has the potential to enhance affinity and/or functional activity at \u03b16 subunit-containing neuronal nicotinic receptors (NNRs) and decrease activation of ganglionic nicotinic receptors, depending on the scaffold. The ramifications of this structure\u2013activity relationship are discussed in the context of the design of small molecules targeting smoking cessation.", "date": "2009-08-01", "date_type": "published", "publication": "Bioorganic and Medicinal Chemistry Letters", "volume": "19", "number": "15", "publisher": "Elsevier", "pagerange": "4359-4363", "id_number": "CaltechAUTHORS:20090730-103918926", "issn": "0960-894X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090730-103918926", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA019375" } ] }, "doi": "10.1016/j.bmcl.2009.05.085", "pmcid": "PMC6107347", "primary_object": { "basename": "mmc1.doc", "url": "https://authors.library.caltech.edu/records/6sfz9-yw476/files/mmc1.doc" }, "related_objects": [ { "basename": "nihms-128262.pdf", "url": "https://authors.library.caltech.edu/records/6sfz9-yw476/files/nihms-128262.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Breining, Scott R.; Bencherif, Merouane; et el." }, { "id": "https://authors.library.caltech.edu/records/fsa02-xa194", "eprint_id": 15532, "eprint_status": "archive", "datestamp": "2023-08-21 21:52:01", "lastmod": "2023-10-19 14:32:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Torrice-M-M", "name": { "family": "Torrice", "given": "Michael M." } }, { "id": "Bower-K-S", "name": { "family": "Bower", "given": "Kiowa S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Probing the role of the cation\u2013\u03c0 interaction in the binding sites of GPCRs using unnatural amino acids", "ispublished": "pub", "full_text_status": "public", "keywords": "D2 receptor; fluorination; membrane protein", "note": "\u00a92009 by the National Academy of Sciences. \n\nEdited by Laura L. Kiessling, University of Wisconsin, Madison, WI, and approved May 1, 2009 (received for review March 24, 2009). Published online before print July 6, 2009, doi: 10.1073/pnas.0903260106 \n\nWe thank C. Doupnik for advice and plasmids and A. Kovoor for helpful discussions. This work was supported by National Institutes of Health Grants GM 081662 and NS011756. \n\nAuthor contributions: M.M.T., K.S.B., H.A.L., and D.A.D. designed research; M.M.T. and K.S.B. performed research; M.M.T., K.S.B., H.A.L., and D.A.D. analyzed data; and M.M.T., K.S.B., and D.A.D. wrote the paper. \n\nThe authors declare no conflict of interest. \n\nThis article is a PNAS Direct Submission. \n\nThis article contains supporting information online at www.pnas.org/cgi/content/full/0903260106/DCSupplemental.\n\nPublished - Torrice2009p5124P_Natl_Acad_Sci_Usa.pdf
Supplemental Material - Torrice0903260106SI.pdf
", "abstract": "We describe a general application of the nonsense suppression methodology for unnatural amino acid incorporation to probe drug\u2013receptor interactions in functional G protein-coupled receptors (GPCRs), evaluating the binding sites of both the M2 muscarinic acetylcholine receptor and the D2 dopamine receptor. Receptors were expressed in Xenopus oocytes, and activation of a G protein-coupled, inward-rectifying K^+ channel (GIRK) provided, after optimization of conditions, a quantitative readout of receptor function. A number of aromatic amino acids thought to be near the agonist-binding site were evaluated. Incorporation of a series of fluorinated tryptophan derivatives at W6.48 of the D2 receptor establishes a cation\u2013\u03c0 interaction between the agonist dopamine and W6.48, suggesting a reorientation of W6.48 on agonist binding, consistent with proposed \"rotamer switch\" models. Interestingly, no comparable cation\u2013\u03c0 interaction was found at the aligning residue in the M2 receptor.", "date": "2009-07-21", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "106", "number": "29", "publisher": "National Academy of Sciences", "pagerange": "11919-11924", "id_number": "CaltechAUTHORS:20090901-141204665", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090901-141204665", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM 081662" }, { "agency": "NIH", "grant_number": "NS011756" } ] }, "doi": "10.1073/pnas.0903260106", "pmcid": "PMC2715518", "primary_object": { "basename": "Torrice0903260106SI.pdf", "url": "https://authors.library.caltech.edu/records/fsa02-xa194/files/Torrice0903260106SI.pdf" }, "related_objects": [ { "basename": "Torrice2009p5124P_Natl_Acad_Sci_Usa.pdf", "url": "https://authors.library.caltech.edu/records/fsa02-xa194/files/Torrice2009p5124P_Natl_Acad_Sci_Usa.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Torrice, Michael M.; Bower, Kiowa S.; et el." }, { "id": "https://authors.library.caltech.edu/records/q7v6b-wq858", "eprint_id": 14807, "eprint_status": "archive", "datestamp": "2023-08-21 21:29:19", "lastmod": "2023-10-18 19:55:22", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Gleitsman-K-R", "name": { "family": "Gleitsman", "given": "Kristin R." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Probing the role of backbone hydrogen bonding in a critical \u03b2 sheet of the extracellular domain of a cys-loop receptor", "ispublished": "pub", "full_text_status": "public", "keywords": "allosterism; backbone esters; ion channels; mutagenesis; receptors", "note": "\u00a9 2009 John Wiley & Sons. \n\nReceived: February 20, 2009. Published online on April 29, 2009. \n\nWe thank Dr. Rigo Pantoja for advice and assistance with TIRF measurements. This work was supported by the NIH (NS 34407; NS 11 756). K.R.G. was partially supported by an NSF Graduate Research Fellowship.\n\nAccepted Version - nihms149703.pdf
", "abstract": "Long-range communication is essential for the function of members of the Cys-loop family of neurotransmitter-gated ion channels. The involvement of the peptide backbone in binding-induced conformational changes that lead to channel gating in these membrane proteins is an interesting, but unresolved issue. To probe the role of the peptide backbone, we incorporated a series of \u03b1-hydroxy acid analogues into the \u03b2-sheet-rich extracellular domain of the muscle subtype of the nicotinic acetylcholine receptor, the prototypical Cys-loop receptor. Specifically, mutations were made in \u03b2 strands 7 and 10 of the \u03b1 subunit. A number of single backbone mutations in this region were well tolerated. However, simultaneous introduction of two proximal backbone mutations led to surface-expressed, nonfunctional receptors. Together, these data suggest that while the receptor is remarkably robust in its ability to tolerate single amide-to-ester mutations throughout these strands, more substantial perturbations to this region have a profound effect on the protein. These results support a model in which backbone movements in the outer sheet are important for receptor function.", "date": "2009-05-25", "date_type": "published", "publication": "ChemBioChem", "volume": "10", "number": "8", "publisher": "Wiley", "pagerange": "1385-1391", "id_number": "CaltechAUTHORS:20090805-085419387", "issn": "1439-4227", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090805-085419387", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "NSF Graduate Research Fellowship" } ] }, "doi": "10.1002/cbic.200900092", "pmcid": "PMC2789490", "primary_object": { "basename": "nihms149703.pdf", "url": "https://authors.library.caltech.edu/records/q7v6b-wq858/files/nihms149703.pdf" }, "resource_type": "article", "pub_year": "2009", "author_list": "Gleitsman, Kristin R.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/easd2-ydy89", "eprint_id": 15826, "eprint_status": "archive", "datestamp": "2023-08-20 01:35:54", "lastmod": "2023-10-19 17:17:23", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Son-C-D", "name": { "family": "Son", "given": "Cagdas D." } }, { "id": "Moss-F-J", "name": { "family": "Moss", "given": "Fraser J." }, "orcid": "0000-0002-8519-6991" }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Nicotine Normalizes Intracellular Subunit Stoichiometry of Nicotinic Receptors Carrying Mutations Linked to Autosomal Dominant Nocturnal Frontal Lobe Epilepsy", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2009 by the American Society for Pharmacology and Experimental Therapeutics.\nReceived January 2, 2009; accepted February 23, 2009.\nThis work was supported by the National Institutes of Health National\nInstitute of Neurological Disorders and Stroke [Grant NS11756]; by Targacept,\nInc.; by a fellowship from Philip Morris USA/International (to C.D.G.); and by\na fellowship from the American Heart Association (to F.J.M.).\nWe thank Princess Imoukhuede, Rigo Pantoja, Rahul Srinivasan,\nLarry Wade, and Ben Corry (University of Western Australia) for\ndiscussion and Jeff Larsen (Nikon) for much technical help.\n\nSupplemental Material - Son2009p2213Mol_Pharmacol_supp_figure.ppt
Supplemental Material - Son2009p2213Mol_Pharmacol_supp_table.pdf
", "abstract": "Autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE)\nis linked with high penetrance to several distinct nicotinic receptor\n(nAChR) mutations. We studied (\u03b14)_3(2\u03b2)_2 versus\n(\u03b14)_2(\u03b22)_3 subunit stoichiometry for five channel-lining M2 domain\nmutations: S247F, S252L, 776ins3 in \u03b14, V287L, and\nV287M in \u03b22. \u03b14 and \u03b22 subunits were constructed with all\npossible combinations of mutant and wild-type (WT) M2 regions,\nof cyan and yellow fluorescent protein, and of fluorescent\nand nonfluorescent M3-M4 loops. Sixteen fluorescent\nsubunit combinations were expressed in N2a cells. Forster\nresonance energy transfer (FRET) was analyzed by donor recovery\nafter acceptor photobleaching and by pixel-by-pixel\nsensitized emission, with confirmation by fluorescence intensity\nratios. Because FRET efficiency is much greater for adjacent\nthan for nonadjacent subunits and the \u03b14 and \u03b22 subunits\noccupy specific positions in nAChR pentamers, observed FRET\nefficiencies from (\u03b14)_3(\u03b22)_2 carrying fluorescent \u03b14 subunits\nwere significantly higher than for (\u03b14)_2(\u03b22)_3; the converse was\nfound for fluorescent 2 subunits. All tested ADNFLE mutants\nproduced 10 to 20% increments in the percentage of intracellular\n(\u03b14)_3(\u03b22)_2 receptors compared with WT subunits. In contrast,\n24- to 48-h nicotine (1 \u00b5M) exposure increased the proportion\nof (\u03b14)_2(\u03b22)_3 in WT receptors and also returned subunit\nstoichiometry to WT levels for \u03b14S248F and \u03b22V287L nAChRs.\nThese observations may be relevant to the decreased seizure\nfrequency in patients with ADNFLE who use tobacco products\nor nicotine patches. Fluorescence-based investigations of\nnAChR subunit stoichiometry may provide efficient drug discovery\nmethods for nicotine addiction or for other disorders\nthat result from dysregulated nAChRs.", "date": "2009-05", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "75", "number": "5", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "1137-1148", "id_number": "CaltechAUTHORS:20090914-084414850", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090914-084414850", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "Targacept" }, { "agency": "Philip Morris" }, { "agency": "American Heart Association" } ] }, "doi": "10.1124/mol.108.054494", "pmcid": "PMC2672806", "primary_object": { "basename": "Son2009p2213Mol_Pharmacol_supp_figure.ppt", "url": "https://authors.library.caltech.edu/records/easd2-ydy89/files/Son2009p2213Mol_Pharmacol_supp_figure.ppt" }, "related_objects": [ { "basename": "Son2009p2213Mol_Pharmacol_supp_table.pdf", "url": "https://authors.library.caltech.edu/records/easd2-ydy89/files/Son2009p2213Mol_Pharmacol_supp_table.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Son, Cagdas D.; Moss, Fraser J.; et el." }, { "id": "https://authors.library.caltech.edu/records/avhdv-n1y52", "eprint_id": 15360, "eprint_status": "archive", "datestamp": "2023-08-20 01:30:54", "lastmod": "2023-10-18 21:42:47", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Gleitsman-K-R", "name": { "family": "Gleitsman", "given": "Kristin R." } }, { "id": "Shanata-J-A-P", "name": { "family": "Shanata", "given": "Jai A. P." } }, { "id": "Frazier-S-J", "name": { "family": "Frazier", "given": "Shawnalea J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Long-Range Coupling in an Allosteric Receptor Revealed by Mutant Cycle Analysis", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2009 by the Biophysical Society. Received 18 September 2008; accepted 15 December 2008. We thank B. N. Cohen for advice on single-channel recording and analysis.\nThis work was supported by the National Institutes of Health (NS 34407; NS\n11756). J.A.P.S. was partially supported by a National Research Service\nAward training grant. K.R.G. was partially supported by a National Science\nFoundation Graduate Research Fellowship.\n\nPublished - Gleitsman2009p4490Biophys_J.pdf
Supplemental Material - mmc1.pdf
", "abstract": "The functional coupling of residues that are far apart in space is the quintessential property of allosteric proteins. For example, in Cys-loop receptors, the gating of an intrinsic ion channel is allosterically regulated by the binding of small molecule neurotransmitters 50\u201360 \u00c5 from the channel gate. Some residues near the binding site must have as their primary function the communication of the binding event to the gating region. These gating pathway residues are essential to function, but their identification and characterization can be challenging. This work introduces a simple strategy, derived from mutant cycle analysis, for identifying gating pathway residues using macroscopic measurements alone. In the exemplar Cys-loop receptor, the nicotinic acetylcholine receptor, a well-characterized reporter mutation (\u03b2L9\u2032S) known to impact gating, was combined with mutations of target residues in the ligand-binding domain hypothesized or previously found to be functionally significant. A mutant cycle analysis of the macroscopic EC50 measurements can then provide insights into the role of the target residue. This new method, elucidating long-range functional coupling in allosteric receptors, can be applied to several reporter mutations in a wide variety of receptors to identify previously characterized and novel mutations that impact the gating pathway. We support our interpretation of macroscopic data with single-channel studies. Elucidating long-range functional coupling in allosteric receptors should be broadly applicable to determining functional roles of residues in allosteric receptors.", "date": "2009-04-22", "date_type": "published", "publication": "Biophysical Journal", "volume": "96", "number": "8", "publisher": "Elsevier", "pagerange": "3168-3178", "id_number": "CaltechAUTHORS:20090827-132541833", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090827-132541833", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NSF" } ] }, "doi": "10.1016/j.bpj.2008.12.3949", "pmcid": "PMC2718292", "primary_object": { "basename": "mmc1.pdf", "url": "https://authors.library.caltech.edu/records/avhdv-n1y52/files/mmc1.pdf" }, "related_objects": [ { "basename": "Gleitsman2009p4490Biophys_J.pdf", "url": "https://authors.library.caltech.edu/records/avhdv-n1y52/files/Gleitsman2009p4490Biophys_J.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Gleitsman, Kristin R.; Shanata, Jai A. P.; et el." }, { "id": "https://authors.library.caltech.edu/records/xqkca-byq14", "eprint_id": 15562, "eprint_status": "archive", "datestamp": "2023-09-25 20:51:56", "lastmod": "2023-10-23 23:31:45", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Imoukhuede-P-I", "name": { "family": "Imoukhuede", "given": "P. I." } }, { "id": "Moss-Fraser-J", "name": { "family": "Moss", "given": "Fraser J." }, "orcid": "0000-0002-8519-6991" }, { "id": "Michael-Darren-J", "name": { "family": "Michael", "given": "Darren J." } }, { "id": "Chow-Robert-H", "name": { "family": "Chow", "given": "Robert H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Ezrin Mediates Tethering of the \u03b3-Aminobutyric Acid Transporter GAT1 to Actin Filaments Via a C-Terminal PDZ-Interacting Domain", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2009 by the Biophysical Society. Received 3 July 2008; accepted 25 November 2008. Editor: Elliot L. Elson.. Available online 2 April 2009. We thank Michael Quick and Elaine Bearer for valuable discussions.\nThis research was supported by grants from the National Institutes of Health\n(DA-09121, DK-60623, and NS-11756), by an American Heart Association\nPostdoctoral Fellowship to F.M., and by the Millard and Muriel Jacobs\nGenetics and Genomics Laboratory at the California Institute of Technology.\n\nPublished - Imoukhuede2009p4492Biophys_J.pdf
Supplemental Material - mmc1.pdf
", "abstract": "A high density of neurotransmitter transporters on axons and presynaptic boutons is required for the efficient clearance of neurotransmitters from the synapse. Therefore, regulators of transporter trafficking (insertion, retrieval, and confinement) can play an important role in maintaining the transporter density necessary for effective function. We determined the interactions that confine GAT1 at the membrane by investigating the lateral mobility of GAT1-yellow fluorescent protein-8 (YFP8) expressed in neuroblastoma 2a cells. Through fluorescence recovery after photobleaching, we found that a significant fraction (~50%) of membrane-localized GAT1 is immobile on the time scale investigated (~150 s). The mobility of the transporter can be increased by depolymerizing actin or by interrupting the GAT1 postsynaptic density 95/Discs large/zona occludens 1 (PDZ)-interacting domain. Microtubule depolymerization, in contrast, does not affect GAT1 membrane mobility. We also identified ezrin as a major GAT1 adaptor to actin. F\u00f6rster resonance energy transfer suggests that GAT1-YFP8 and cyan fluorescent (CFP) tagged ezrin (ezrin-CFP) exist within a complex that has a F\u00f6rster resonance energy transfer efficiency of 19% \u00b1 2%. This interaction can be diminished by disrupting the actin cytoskeleton. In addition, the disruption of actin results in a >3-fold increase in \u03b3-aminobutyric acid uptake, apparently via a mechanism distinct from the PDZ-interacting protein. Our data reveal that actin confines GAT1 to the plasma membrane via ezrin, and this interaction is mediated through the PDZ-interacting domain of GAT1.", "date": "2009-04-08", "date_type": "published", "publication": "Biophysical Journal", "volume": "96", "number": "7", "publisher": "Biophysical Society", "pagerange": "2949-2960", "id_number": "CaltechAUTHORS:20090903-090332845", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090903-090332845", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "DK-60623" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "American Heart Association" }, { "agency": "Millard and Muriel Jacobs Genetics and Genomics Laboratory" } ] }, "local_group": { "items": [ { "id": "Millard-and-Muriel-Jacobs-Genetics-and-Genomics-Laboratory" } ] }, "doi": "10.1016/j.bpj.2008.11.070", "pmcid": "PMC2711277", "primary_object": { "basename": "mmc1.pdf", "url": "https://authors.library.caltech.edu/records/xqkca-byq14/files/mmc1.pdf" }, "related_objects": [ { "basename": "Imoukhuede2009p4492Biophys_J.pdf", "url": "https://authors.library.caltech.edu/records/xqkca-byq14/files/Imoukhuede2009p4492Biophys_J.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Imoukhuede, P. I.; Moss, Fraser J.; et el." }, { "id": "https://authors.library.caltech.edu/records/jhf8y-q4g06", "eprint_id": 14041, "eprint_status": "archive", "datestamp": "2023-08-21 21:04:38", "lastmod": "2023-10-18 16:03:58", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Xiu-Xinan", "name": { "family": "Xiu", "given": "Xinan" } }, { "id": "Puskar-N-L", "name": { "family": "Puskar", "given": "Nyssa L." } }, { "id": "Shanata-J-A-P", "name": { "family": "Shanata", "given": "Jai A. P." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Nicotine binding to brain receptors requires a strong cation\u2013\u03c0 interaction", "ispublished": "pub", "full_text_status": "public", "keywords": "acetylcholine-receptors; ion channels; M2 domain; agonists; ACHBP; site; activation; tolerance; complexes; chemistry", "note": "\u00a9 2009 Nature Publishing Group.\n\nReceived 26 November 2008; Accepted 9 January 2009; Published online 1 March 2009.\n\nWe thank B. N. Cohen for advice on single-channel recording and analysis. This work was supported by the NIH (NS 34407; NS 11756) and the California Tobacco-Related Disease Research Program of the University of California, grant number 16RT-0160. J.A.P.S. was partially supported by an NRSA training grant.\n\nAccepted Version - nihms103025.pdf
Supplemental Material - Xiu2009p1230Naturesupp.pdf
", "abstract": "Nicotine addiction begins with high-affinity binding of nicotine to\nacetylcholine (ACh) receptors in the brain. The end result is over\n4,000,000 smoking-related deaths annually worldwide and the\nlargest source of preventable mortality in developed countries.\nStress reduction, pleasure, improved cognition and other central\nnervous system effects are strongly associated with smoking.\nHowever, if nicotine activated ACh receptors found in muscle as\npotently as it does brain ACh receptors, smoking would cause intolerable\nand perhaps fatal muscle contractions. Despite extensive\npharmacological, functional and structural studies of ACh receptors,\nthe basis for the differential action of nicotine on brain compared\nwith muscle ACh receptors has not been determined. Here we\nshow that at the \u03b14\u03b22 brain receptors thought to underlie nicotine\naddiction, the high affinity for nicotine is the result of a strong\ncation\u2013\u03c0 interaction to a specific aromatic amino acid of the receptor,\nTrpB. In contrast, the low affinity for nicotine at the muscle type ACh\nreceptor is largely due to the fact that this key interaction is\nabsent, even though the immediate binding site residues, including\nthe key amino acid TrpB, are identical in the brain and muscle\nreceptors. At the same time a hydrogen bond from nicotine to the\nbackbone carbonyl of TrpB is enhanced in the neuronal receptor\nrelative to the muscle type. A point mutation near TrpB that differentiates\n\u03b14\u03b22 and muscle-type receptors seems to influence the\nshape of the binding site, allowing nicotine to interact more strongly\nwith TrpB in the neuronal receptor. ACh receptors are established\ntherapeutic targets for Alzheimer's disease, schizophrenia,\nParkinson's disease, smoking cessation, pain, attention-deficit\nhyperactivity disorder, epilepsy, autism and depression. Along with\nsolving a chemical mystery in nicotine addiction, our results provide\nguidance for efforts to develop drugs that target specific types of\nnicotinic receptors.", "date": "2009-03-26", "date_type": "published", "publication": "Nature", "volume": "458", "number": "7237", "publisher": "Nature Publishing Group", "pagerange": "534-537", "id_number": "CaltechAUTHORS:20090422-090708342", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090422-090708342", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "16RT-0160" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1038/nature07768", "pmcid": "PMC2755585", "primary_object": { "basename": "Xiu2009p1230Naturesupp.pdf", "url": "https://authors.library.caltech.edu/records/jhf8y-q4g06/files/Xiu2009p1230Naturesupp.pdf" }, "related_objects": [ { "basename": "nihms103025.pdf", "url": "https://authors.library.caltech.edu/records/jhf8y-q4g06/files/nihms103025.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Xiu, Xinan; Puskar, Nyssa L.; et el." }, { "id": "https://authors.library.caltech.edu/records/w9v7t-c1f68", "eprint_id": 14612, "eprint_status": "archive", "datestamp": "2023-08-20 01:03:03", "lastmod": "2023-10-18 18:08:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Bohland-J", "name": { "family": "Bohland", "given": "Jason" } }, { "id": "Wu-Caizhi", "name": { "family": "Wu", "given": "Caizhi" } }, { "id": "Barbas-H", "name": { "family": "Barbas", "given": "Helen" } }, { "id": "Bokil-H", "name": { "family": "Bokil", "given": "Hemant" } }, { "id": "Bota-M", "name": { "family": "Bota", "given": "Mihail" } }, { "id": "Breiter-H-C", "name": { "family": "Breiter", "given": "Hans C." } }, { "id": "Cline-H-T", "name": { "family": "Cline", "given": "Hollis T." } }, { "id": "Doyle-J-C", "name": { "family": "Doyle", "given": "John C." }, "orcid": "0000-0002-1828-2486" }, { "id": "Freed-P-J", "name": { "family": "Freed", "given": "Peter J." } }, { "id": "Greenspan-R-J", "name": { "family": "Greenspan", "given": "Ralph J." }, "orcid": "0000-0002-6787-2845" }, { "id": "Haber-S-N", "name": { "family": "Haber", "given": "Suzanne N." } }, { "id": "Hawrylycz-M", "name": { "family": "Hawrylycz", "given": "Michael" } }, { "id": "Herrera-D-G", "name": { "family": "Herrera", "given": "Daniel G." } }, { "id": "Hilgetag-C-C", "name": { "family": "Hilgetag", "given": "Claus C." } }, { "id": "Huang-Z-Josh", "name": { "family": "Huang", "given": "Z. Josh" } }, { "id": "Jones-A", "name": { "family": "Jones", "given": "Allan" } }, { "id": "Jones-E-G", "name": { "family": "Jones", "given": "Edward G." } }, { "id": "Karten-H-J", "name": { "family": "Karten", "given": "Harvey J." } }, { "id": "Kleinfeld-D", "name": { "family": "Kleinfeld", "given": "David" } }, { "id": "K\u00f6tter-R", "name": { "family": "K\u00f6tter", "given": "Rolf" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Lin-John-M", "name": { "family": "Lin", "given": "John M." } }, { "id": "Mensh-B-D", "name": { "family": "Mensh", "given": "Brett D." } }, { "id": "Mikula-S", "name": { "family": "Mikula", "given": "Shawn" } }, { "id": "Panskepp-J", "name": { "family": "Panskepp", "given": "Jaak" } }, { "id": "Price-J-L", "name": { "family": "Price", "given": "Joseph L." } }, { "id": "Safdieh-J", "name": { "family": "Safdieh", "given": "Joseph" } }, { "id": "Saper-C-B", "name": { "family": "Saper", "given": "Clifford B." } }, { "id": "Schiff-N-D", "name": { "family": "Schiff", "given": "Nicholas D." } }, { "id": "Schmahmann-J-D", "name": { "family": "Schmahmann", "given": "Jeremy D." } }, { "id": "Stillman-B-W", "name": { "family": "Stillman", "given": "Bruce W." } }, { "id": "Svoboda-K", "name": { "family": "Svoboda", "given": "Karel" } }, { "id": "Swanson-L-W", "name": { "family": "Swanson", "given": "Larry W." } }, { "id": "Toga-A-W", "name": { "family": "Toga", "given": "Arthur W." } }, { "id": "Van-Essen-D-C", "name": { "family": "Van Essen", "given": "David C." }, "orcid": "0000-0001-7044-4721" }, { "id": "Watson-J-D", "name": { "family": "Watson", "given": "James D." } }, { "id": "Mitra-P-P", "name": { "family": "Mitra", "given": "Partha P." } } ] }, "title": "A proposal for a coordinated effort for the determination of brainwide neuroanatomical connectivity in model organisms at a mesoscopic scale", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2009 Bohland et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.\nPublished March 27, 2009.\nThis paper is a result of discussions at the 2007 and 2008 Brain\nArchitecture Project Banbury Center Meetings, funded by the W. M. Keck\nFoundation. The sponsors had no role in the conception or preparation of this\nmanuscript.\n\nPublished - Bohland2009p4251Plos_Comput_Biol.pdf
", "abstract": "In this era of complete genomes, our knowledge of neuroanatomical circuitry remains surprisingly sparse. Such knowledge is critical, however, for both basic and clinical research into brain function. Here we advocate for a concerted effort to fill this gap, through systematic, experimental mapping of neural circuits at a mesoscopic scale of resolution suitable for comprehensive, brainwide coverage, using injections of tracers or viral vectors. We detail the scientific and medical rationale and briefly review existing knowledge and experimental techniques. We define a set of desiderata, including brainwide coverage; validated and extensible experimental techniques suitable for standardization and automation; centralized, open-access data repository; compatibility with existing resources; and tractability with current informatics technology. We discuss a hypothetical but tractable plan for mouse, additional efforts for the macaque, and technique development for human. We estimate that the mouse connectivity project could be completed within five years with a comparatively modest budget.", "date": "2009-03", "date_type": "published", "publication": "PLoS Computational Biology", "volume": "5", "number": "3", "publisher": "Public Library of Science", "pagerange": "Art. No. e1000334", "id_number": "CaltechAUTHORS:20090717-142050047", "issn": "1553-734X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090717-142050047", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1371/journal.pcbi.1000334", "pmcid": "PMC2655718", "primary_object": { "basename": "Bohland2009p4251Plos_Comput_Biol.pdf", "url": "https://authors.library.caltech.edu/records/w9v7t-c1f68/files/Bohland2009p4251Plos_Comput_Biol.pdf" }, "resource_type": "article", "pub_year": "2009", "author_list": "Bohland, Jason; Wu, Caizhi; et el." }, { "id": "https://authors.library.caltech.edu/records/9x8d1-c0d65", "eprint_id": 14504, "eprint_status": "archive", "datestamp": "2023-08-21 20:54:52", "lastmod": "2023-10-18 18:03:38", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Srinivasan-R", "name": { "family": "Srinivasan", "given": "Rahul" } }, { "id": "Son-C-D", "name": { "family": "Son", "given": "Cagdas D." } }, { "id": "Miwa-Julie-M", "name": { "family": "Miwa", "given": "Julie" } }, { "id": "Pantoja-R", "name": { "family": "Pantoja", "given": "Rigo" } }, { "id": "Banghart-M-R", "name": { "family": "Banghart", "given": "Matthew R." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Goate-A-M", "name": { "family": "Goate", "given": "Alison M." } }, { "id": "Wang-Jen-C", "name": { "family": "Wang", "given": "Jen C." } } ] }, "title": "Nicotine is a Selective Pharmacological Chaperone of Acetylcholine Receptor Number and Stoichiometry. Implications for Drug Discovery", "ispublished": "pub", "full_text_status": "restricted", "keywords": "ADNFLE; dopamine; GABA; proteostasis; upregulation", "note": "\u00a9 2009 American Association of Pharmaceutical Scientists. \n\nReceived 31 December 2008; accepted 7 February 2009; published online 12 March 2009. \n\nRelevant work in the authors' laboratories is supported\nby grants from NINDS (NS11756, NS34407), NIDA\n(DA17279, DA19375), NCI (CA089392), NIA (AG033954),\nNIAAA (AA08401), the Michael J. Fox Foundation, the\nCalifornia Tobacco-Related Disease Research Program, the\nCroll Autism Research Foundation (1004564-01-CEN5300443), and Targacept, Inc. Postdoctoral fellowships\nwere awarded to C. S. from Philip Morris USA/International,\nto X. C. from the California Tobacco-Related Disease\nResearch Program, and to R. P. from the Ford Foundation\nand APA-DPN.", "abstract": "The acronym SePhaChARNS, for \"selective pharmacological chaperoning of acetylcholine\nreceptor number and stoichiometry,\" is introduced. We hypothesize that SePhaChARNS underlies\nclassical observations that chronic exposure to nicotine causes \"upregulation\" of nicotinic receptors\n(nAChRs). If the hypothesis is proven, (1) SePhaChARNS is the molecular mechanism of the first step in\nneuroadaptation to chronic nicotine; and (2) nicotine addiction is partially a disease of excessive\nchaperoning. The chaperone is a pharmacological one, nicotine; and the chaperoned molecules are\n\u03b14\u03b22* nAChRs. SePhaChARNS may also underlie two inadvertent therapeutic effects of tobacco use:\n(1) the inverse correlation between tobacco use and Parkinson's disease; and (2) the suppression of\nseizures by nicotine in autosomal dominant nocturnal frontal lobe epilepsy. SePhaChARNS arises from\nthe thermodynamics of pharmacological chaperoning: ligand binding, especially at subunit interfaces,\nstabilizes AChRs during assembly and maturation, and this stabilization is most pronounced for the\nhighest-affinity subunit compositions, stoichiometries, and functional states of receptors. Several chemical\nand pharmacokinetic characteristics render exogenous nicotine a more potent pharmacological\nchaperone than endogenous acetylcholine. SePhaChARNS is modified by desensitized states of nAChRs,\nby acid trapping of nicotine in organelles, and by other aspects of proteostasis. SePhaChARNS is\nselective at the cellular, and possibly subcellular, levels because of variations in the detailed nAChR\nsubunit composition, as well as in expression of auxiliary proteins such as lynx. One important\nimplication of the SePhaChARNS hypothesis is that therapeutically relevant nicotinic receptor drugs\ncould be discovered by studying events in intracellular compartments rather than exclusively at the\nsurface membrane.", "date": "2009-03", "date_type": "published", "publication": "AAPS Journal", "volume": "11", "number": "1", "publisher": "American Association of Pharmaceutical Scientists", "pagerange": "167-177", "id_number": "CaltechAUTHORS:20090707-084125313", "issn": "1550-7416", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090707-084125313", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "AG033954" }, { "agency": "NIH", "grant_number": "AA08401" }, { "agency": "Michael J. Fox Foundation" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Croll Autism Research Foundation", "grant_number": "1004564-01CEN5300443" }, { "agency": "Targacept, Inc." }, { "agency": "Philip Morris USA/ International" }, { "agency": "Ford Foundation" }, { "agency": "American Psychological Association" }, { "agency": "NIH", "grant_number": "CA089392" }, { "agency": "National Institute of Neurological Disorders and Stroke (NINDS)" }, { "agency": "National Cancer Institute" }, { "agency": "National Institute on Aging" }, { "agency": "National Institute on Alcohol Abuse and Alcoholism" } ] }, "doi": "10.1208/s12248-009-9090-7", "pmcid": "PMC2664890", "resource_type": "article", "pub_year": "2009", "author_list": "Lester, Henry A.; Xiao, Cheng; et el." }, { "id": "https://authors.library.caltech.edu/records/615d9-1hz47", "eprint_id": 15688, "eprint_status": "archive", "datestamp": "2023-08-20 00:40:33", "lastmod": "2023-10-19 14:43:37", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Pantoja-R", "name": { "family": "Pantoja", "given": "Rigo" } }, { "id": "Rodriguez-E-A", "name": { "family": "Rodriguez", "given": "Erik A." } }, { "id": "Dibas-M-I", "name": { "family": "Dibas", "given": "Mohammed I." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Single-Molecule Imaging of a Fluorescent Unnatural Amino Acid Incorporated Into Nicotinic Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2009 Biophysical Society.\nSubmitted February 29, 2008, and accepted for publication September 29,\n2008.\nWe gratefully acknowledge the assistance of Dr. E. James Petersson at the\noutset of this project. We thank Dr. Shelly Tzlil for single-molecule discussions,\nand for help with programming. Mr. Edward Wang provided assistance\nwith ImageJ Java programming and data documentation. We also\nthank the members of the Lester and Dougherty Laboratory Group for\ninvaluable discussions.\nR.P. was supported by postdoctoral fellowships from the Ford Foundation\nand American Psychological Association Diversity Program in Neuroscience.\nM.I.D. was supported by a National Research Service Award from\nthe National Institutes of Health. E.A.R. is a National Science Foundation\nPredoctoral Fellow. This work was supported by National Institutes of\nHealth grants NS 11756, NS 34407, and HL 79350. \n\nTwo figures are available at http://www.biophysj.org/biophysj/supplemental/S0006-3495(08)00012-X.\n\nPublished - Pantoja2009p4493Biophys_J.pdf
Supplemental Material - Pantoja2009p4493Biophys_J_supp.pdf
", "abstract": "We report on the first, to our knowledge, successful detection of a fluorescent unnatural amino acid (fUAA), Lys(BODIPYFL), incorporated into a membrane protein (the muscle nicotinic acetylcholine receptor, nAChR) in a living cell. Xenopus oocytes were injected with a frameshift-suppressor tRNA, amino-acylated with Lys(BODIPYFL) and nAChR (\u03b1/\u03b219\u2032GGGU/\u03b3/\u03b4) mRNAs. We measured fluorescence from oocytes expressing nAChR \u03b219\u2032Lys(BODIPYFL), using time-resolved total internal reflection fluorescence microscopy. Under conditions of relatively low receptor density (<0.1 receptors/\u03bcm^2), we observed puncta with diffraction-limited profiles that were consistent with the point-spread function of our microscope. Furthermore, diffraction-limited puncta displayed step decreases in fluorescence intensity, consistent with single-molecule photobleaching. The puncta densities agreed with macroscopic ACh-induced current densities, showing that the fUAA was incorporated, and that receptors were functional. Dose-response relations for the nAChR \u03b219\u2032Lys(BODIPYFL) receptors were similar to those for wild-type receptors. We also studied nAChR \u03b219\u2032Lys(BODIPYFL) receptors labeled with \u03b1-bungarotoxin monoconjugated with Alexa488 (\u03b1BtxAlexa488). The nAChR has two \u03b1Btx binding sites, and puncta containing the Lys(BODIPYFL) labeled with \u03b1BtxAlexa488 yielded the expected three discrete photobleaching steps. We also performed positive control experiments with a nAChR containing enhanced green fluorescent protein in the \u03b3-subunit M3-M4 loop, which confirmed our nAChR \u03b219\u2032Lys(BODIPYFL) measurements. Thus, we report on the cell-based single-molecule detection of nAChR \u03b219\u2032Lys(BODIPYFL).", "date": "2009-01-07", "date_type": "published", "publication": "Biophysical Journal", "volume": "96", "number": "1", "publisher": "Biophysical Society", "pagerange": "226-237", "id_number": "CaltechAUTHORS:20090909-105237809", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20090909-105237809", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Ford Foundation" }, { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "HL 79350" }, { "agency": "American Psychological Association" }, { "agency": "NSF" } ] }, "doi": "10.1016/j.bpj.2008.09.034", "pmcid": "PMC2710013", "primary_object": { "basename": "Pantoja2009p4493Biophys_J.pdf", "url": "https://authors.library.caltech.edu/records/615d9-1hz47/files/Pantoja2009p4493Biophys_J.pdf" }, "related_objects": [ { "basename": "Pantoja2009p4493Biophys_J_supp.pdf", "url": "https://authors.library.caltech.edu/records/615d9-1hz47/files/Pantoja2009p4493Biophys_J_supp.pdf" } ], "resource_type": "article", "pub_year": "2009", "author_list": "Pantoja, Rigo; Rodriguez, Erik A.; et el." }, { "id": "https://authors.library.caltech.edu/records/6qhtj-40r39", "eprint_id": 12183, "eprint_status": "archive", "datestamp": "2023-08-22 13:53:42", "lastmod": "2023-10-17 16:35:06", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Gleitsman-K-R", "name": { "family": "Gleitsman", "given": "Kristin Rule" } }, { "id": "Kedrowski-S-M-A", "name": { "family": "Kedrowski", "given": "Sean M. A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "An intersubunit hydrogen bond in the nicotinic acetylcholine receptor that contributes to channel gating", "ispublished": "pub", "full_text_status": "public", "note": "Copyright 2008 by The American Society for Biochemistry and Molecular Biology, Inc. \n\nReceived for publication, September 17, 2008, and in revised form, October 21, 2008. \n\nThis work was supported, in whole or in part, by National Institutes of Health Grants NS 34407 and NS 11756. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. \n\nThe online version of this article (available at http://www.jbc.org) contains supplemental text, a supplemental table, and a supplemental figure. \n\nThese authors [K.R.G., S.M.A.K.] contributed equally to this work. \n\n[K.R.G. was] [p]artially supported by a National Science Foundation graduate research fellowship. \n\n[S.M.A.K. was] [p]artially supported by a National Research Service Award training grant.\n\nPublished - GLEjbc08.pdf
Accepted Version - GLEjbc08pip.pdf
Supplemental Material - GLEjbc08supp.pdf
", "abstract": "The muscle nicotinic acetylcholine receptor (nAChR) is a large, allosteric, ligand-gated ion channel with the subunit composition \u03b12\u03b2\u03b3\u03b4. Though much is now known about the structure of the binding site, relatively little is understood about how the binding event is communicated to the channel gate, causing the pore to open. Here we identify a key hydrogen bond near the binding site that is involved in the gating pathway. Using mutant cycle analysis with the novel unnatural residue \u03b1-hydroxyserine (Sah), we find that the backbone N-H of \u03b1S191 in loop C makes a hydrogen bond to an anionic side chain of the complementary subunit upon agonist binding. However, the anionic partner is not the glutamate predicted by the crystal structures of the homologous acetylcholine binding protein (AChBP). Instead, the hydrogen bonding partner is the extensively researched aspartate \u03b3D174/\u03b4D180 -\u2014 which had originally been identified as a key binding residue for cationic agonists.", "date": "2008-12-19", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "283", "number": "51", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "35638-35643", "id_number": "CaltechAUTHORS:GLEjbc08", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:GLEjbc08", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "NSF Graduate Research Fellowship" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1074/jbc.M807226200", "pmcid": "PMC2602911", "primary_object": { "basename": "GLEjbc08.pdf", "url": "https://authors.library.caltech.edu/records/6qhtj-40r39/files/GLEjbc08.pdf" }, "related_objects": [ { "basename": "GLEjbc08pip.pdf", "url": "https://authors.library.caltech.edu/records/6qhtj-40r39/files/GLEjbc08pip.pdf" }, { "basename": "GLEjbc08supp.pdf", "url": "https://authors.library.caltech.edu/records/6qhtj-40r39/files/GLEjbc08supp.pdf" } ], "resource_type": "article", "pub_year": "2008", "author_list": "Gleitsman, Kristin Rule; Kedrowski, Sean M. A.; et el." }, { "id": "https://authors.library.caltech.edu/records/het29-fyx11", "eprint_id": 12182, "eprint_status": "archive", "datestamp": "2023-08-22 13:24:40", "lastmod": "2023-10-17 16:35:04", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Pless-S-A", "name": { "family": "Pless", "given": "Stephan A." } }, { "id": "Millen-K-S", "name": { "family": "Millen", "given": "Kat S." } }, { "id": "Hanek-A-P", "name": { "family": "Hanek", "given": "Ariele P." } }, { "id": "Lynch-J-W", "name": { "family": "Lynch", "given": "Joseph W." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "Sarah C. R." }, "orcid": "0000-0001-9410-9805" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "A Cation-\u03c0 Interaction in the Binding Site of the Glycine Receptor Is Mediated by a Phenylalanine Residue", "ispublished": "pub", "full_text_status": "public", "keywords": "ligand-gated ion channel; Cys-loop receptor; cation-{pi} interaction; unnatural amino acids; nonsense suppression; neurotransmitter", "note": "\u00a9 2008 Society for Neuroscience. \n\nReceived August 6, 2008; accepted August 9, 2008. \n\nThis work was supported by the Wellcome Trust (S.C.R.L. is a Wellcome Trust Senior Research Fellow in Basic Biomedical Science), a Biotechnology and Biological Sciences Research Council studentship (K.S.M.), National Institutes of Health Grants NS11756 and NS34407, the Australian Research Council, a National Health and Medical Research Council of Australia Senior Research Fellowship (J.W.L.), and a University of Queensland International Postgraduate Research Scholarship (S.A.P.). \n\nS.C.R.L. and D.A.D. contributed equally to this work.\n\nPublished - PLEjns08.pdf
", "abstract": "Cys-loop receptor binding sites characteristically contain many aromatic amino acids. In nicotinic ACh and 5-HT3 receptors, a Trp residue forms a cation-{pi} interaction with the agonist, whereas in GABAA receptors, a Tyr performs this role. The glycine receptor binding site, however, contains predominantly Phe residues. Homology models suggest that two of these Phe side chains, Phe159 and Phe207, and possibly a third, Phe63, are positioned such that they could contribute to a cation-{pi} interaction with the primary amine of glycine. Here, we test this hypothesis by incorporation of a series of fluorinated Phe derivatives using unnatural amino acid mutagenesis. The data reveal a clear correlation between the glycine EC50 value and the cation-{pi} binding ability of the fluorinated Phe derivatives at position 159, but not at positions 207 or 63, indicating a single cation-{pi} interaction between glycine and Phe159. The data thus provide an anchor point for locating glycine in its binding site, and demonstrate for the first time a cation-{pi} interaction between Phe and a neurotransmitter.", "date": "2008-10-22", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "28", "number": "43", "publisher": "Society for Neuroscience", "pagerange": "10937-10942", "id_number": "CaltechAUTHORS:PLEjns08", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:PLEjns08", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "Biotechnology and Biological Sciences Research Council (BBSRC)" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "Australian Research Council" }, { "agency": "National Health and Medical Research Council (NHMRC)" }, { "agency": "University of Queensland" } ] }, "doi": "10.1523/JNEUROSCI.2540-08.2008", "pmcid": "PMC2649377", "primary_object": { "basename": "PLEjns08.pdf", "url": "https://authors.library.caltech.edu/records/het29-fyx11/files/PLEjns08.pdf" }, "resource_type": "article", "pub_year": "2008", "author_list": "Pless, Stephan A.; Millen, Kat S.; et el." }, { "id": "https://authors.library.caltech.edu/records/e4mpm-pma13", "eprint_id": 13498, "eprint_status": "archive", "datestamp": "2023-08-22 13:20:01", "lastmod": "2023-10-17 23:56:46", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Grady-S-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Whiteaker-P", "name": { "family": "Whiteaker", "given": "Paul" } }, { "id": "McClure-Begley-T", "name": { "family": "McClure-Begley", "given": "Tristan" } }, { "id": "McKinney-S", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Miwa-Julie-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Bupp-S", "name": { "family": "Bupp", "given": "Sujata" } }, { "id": "Heintz-N", "name": { "family": "Heintz", "given": "Nathaniel" } }, { "id": "McIntosh-J-M", "name": { "family": "McIntosh", "given": "J. Michael" } }, { "id": "Bencherif-M", "name": { "family": "Bencherif", "given": "Merouane" } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "In vivo activation of midbrain dopamine neurons via sensitized, high-affinity \u03b16*nicotinic acetylcholine receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "MOLNEURO; SYSNEURO", "note": "\u00a9 2008 Elsevier B.V.\nAccepted 4 September 2008. Published: October 8, 2008. Available online 8 October 2008. \nWe thank members of the Lester lab for helpful discussions. Special thanks to\nB. Drenan. Thanks to C. Wageman, E. Myers, C. Xiao, A. Tapper, R. Nashmi,\nC. Fonck, J. Schwarz, J. Jankowsky, M. Liu, P. Deshpande, S. Benazouz,\nand C. Zhou. This work was supported by H.H.M.I. (N.H., J.M. Miwa, S.B.)\nand grants from NIH (DA09121, DA17279, and NS11756 to H.A.L.; DA19375\nto H.A.L. and M.J.M.; DA03194 to M.J.M.; DA12242 to M.J.M. and P.W.;\nMH53631 and GM48677 to J.M. McIntosh), the Moore Foundation, the Croll\nResearch Foundation (to J.M. Miwa) and the California Tobacco Related\nDisease Research Program (TRDRP; 12RT-0245 to H.A.L.). R.M.D. was\nsupported by postdoctoral fellowships from TRDRP (15FT-0030) and NIH\n(DA021492 and NS007251).\n\nAccepted Version - nihms75830.pdf
Supplemental Material - DREn08_supp.pdf
", "abstract": "\u03b16-containing (\u03b16*) nicotinic ACh receptors (nAChRs) are selectively expressed in dopamine (DA) neurons and participate in cholinergic transmission. We generated and studied mice with gain-of-function \u03b16* nAChRs, which isolate and amplify cholinergic control of DA transmission. In contrast to gene knockouts or pharmacological blockers, which show necessity, we show that activating \u03b16* nAChRs and DA neurons is sufficient to cause locomotor hyperactivity. \u03b16(L9'S) mice are hyperactive in their home cage and fail to habituate to a novel environment. Selective activation of \u03b16* nAChRs with low doses of nicotine, by stimulating DA but not GABA neurons, exaggerates these phenotypes and produces a hyperdopaminergic state in vivo. Experiments with additional nicotinic drugs show that altering agonist efficacy at \u03b16* provides fine tuning of DA release and locomotor responses. \u03b16*-specific agonists or antagonists may, by targeting endogenous cholinergic mechanisms in midbrain or striatum, provide a method for manipulating DA transmission in neural disorders.", "date": "2008-10-09", "date_type": "published", "publication": "Neuron", "volume": "60", "number": "1", "publisher": "Elsevier", "pagerange": "123-136", "id_number": "CaltechAUTHORS:DREn08", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:DREn08", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA09121" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "DA19375" }, { "agency": "NIH", "grant_number": "DA03194" }, { "agency": "NIH", "grant_number": "DA12242" }, { "agency": "NIH", "grant_number": "MH53631" }, { "agency": "NIH", "grant_number": "GM48677" }, { "agency": "NIH", "grant_number": "DA021492" }, { "agency": "NIH", "grant_number": "NS007251" }, { "agency": "Gordon and Betty Moore Foundation" }, { "agency": "Croll Research Foundation" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "12RT-0245" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "15FT-0030" }, { "agency": "Howard Hughes Medical Institute (HHMI)" } ] }, "doi": "10.1016/j.neuron.2008.09.009", "pmcid": "PMC2632732", "primary_object": { "basename": "DREn08_supp.pdf", "url": "https://authors.library.caltech.edu/records/e4mpm-pma13/files/DREn08_supp.pdf" }, "related_objects": [ { "basename": "nihms75830.pdf", "url": "https://authors.library.caltech.edu/records/e4mpm-pma13/files/nihms75830.pdf" } ], "resource_type": "article", "pub_year": "2008", "author_list": "Drenan, Ryan M.; Grady, Sharon R.; et el." }, { "id": "https://authors.library.caltech.edu/records/detec-g8310", "eprint_id": 12139, "eprint_status": "archive", "datestamp": "2023-08-22 13:19:14", "lastmod": "2023-10-17 16:33:23", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Hanek-A-P", "name": { "family": "Hanek", "given": "Ariele P." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "A Stereochemical Test of a Proposed Structural Feature of the Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2008 American Chemical Society. \n\nReceived March 6, 2008. Web Release Date: September 10, 2008. \n\nWe thank the National Institutes of Health for their generous support (grants NS 34407 and NS 11756 and predoctoral training grant 5T32GM07616). \n\nSupporting Information Available: Experimental procedures for incorporation of unnatural amino acids and electrophysiological evaluation of mutants, dose response curves for mutations discussed herein, sample traces of select mutants, discussion of previously reported conventional mutants, as well as the synthesis and characterization of NVOC-aThr cyano-methyl ester. This material is available free of charge via the Internet at http://pubs.acs.org.\n\nSupplemental Material - ja8015457-File002.pdf
", "abstract": "Understanding the gating mechanism of the nicotinic acetylcholine receptor (nAChR) and similar channels constitutes a significant challenge in chemical neurobiology. In the present work, we use a stereochemical probe to evaluate a proposed pin-into-hydrophobic socket mechanism for the \u03b1Val46 side chain of the nAChR. Utilizing nonsense suppression methodology we incorporated isoleucine (Ile), O-methyl threonine (Omt) and threonine (Thr) as well as their side chain epimers (the allo counterparts). Surprisingly, our results indicate that only the pro-S methyl group of the \u03b1Val46 side chain is sensitive to changes in hydrophobicity, consistent with the precise geometrical requirements of the pin-into-socket mechanism.", "date": "2008-10-08", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "130", "number": "40", "publisher": "American Chemical Society", "pagerange": "13216-13218", "id_number": "CaltechAUTHORS:HANjacs08", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:HANjacs08", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5T32GM07616" } ] }, "doi": "10.1021/ja8015457", "primary_object": { "basename": "ja8015457-File002.pdf", "url": "https://authors.library.caltech.edu/records/detec-g8310/files/ja8015457-File002.pdf" }, "resource_type": "article", "pub_year": "2008", "author_list": "Hanek, Ariele P.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/tjk3d-0dw59", "eprint_id": 13433, "eprint_status": "archive", "datestamp": "2023-08-22 12:48:28", "lastmod": "2023-10-17 23:53:37", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Fonck-C", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Salas-R", "name": { "family": "Salas", "given": "Ramiro" } }, { "id": "Zhou-Chunyi", "name": { "family": "Zhou", "given": "Chunyi" } }, { "id": "Huang-Qi", "name": { "family": "Huang", "given": "Qi" } }, { "id": "De-Biasi-M", "name": { "family": "De Biasi", "given": "Mariella" } }, { "id": "Lester-R-A-J", "name": { "family": "Lester", "given": "Robin A. J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Demonstration of functional \u03b14-containing nicotinic receptors in the medial habenula", "ispublished": "pub", "full_text_status": "public", "keywords": "Addiction; Ion channel; C-fos; Knock-in; mRNA", "note": "\u00a9 2008 Elsevier Ltd.\nReceived 6 August 2008; accepted 8 August 2008; available online 26 August 2008.\nThis work was supported by PHS grants NS31669, DA17173,\nDA17279, California Tobacco-Related Disease Research Project, and the Philip Morris External Research Fund. R.N. was supported by a NARSAD Young Investigator Award.\n\nAccepted Version - nihms87064.pdf
", "abstract": "The medial habenula (MHb) exhibits exceptionally high levels of nicotinic acetylcholine receptors (nAChRs), but it remains unclear whether all expressed nAChR subunit mRNAs are translated to form functional receptors. In particular \u03b14 subunits have not been reported to have any functional role, despite strong \u03b14 mRNA expression in the ventrolateral MHb. We studied a strain of knock-in mice expressing fluorescent \u03b14* nAChRs (\u03b14YFP), as well as a knock-in strain expressing hypersensitive \u03b14* nAChRs (\u03b14L9'A). In \u03b14YFP mice, there was strong fluorescence in the ventrolateral MHb. In hypersensitive \u03b14L9'A mice, injections of a low dose of nicotine (0.1 mg/kg) led to strong c-fos expression in only the ventrolateral region of the MHb, but not in the MHb of wild-type (WT) mice. In MHb slice recordings, ventrolateral neurons from \u03b14L9'A mice, but not from WT mice, responded robustly to nicotine (1 \u03bcM). Neurons in the medial aspect of the MHb had >10-fold smaller responses. Thus \u03b14* nAChRs contribute to the selective activation of a subset of MHb neurons. Subunit composition analysis based on gain-offunction knock-in mice provides a useful experimental paradigm.", "date": "2008-08-26", "date_type": "published", "publication": "Neuropharmacology", "volume": "56", "number": "1", "publisher": "Elsevier", "pagerange": "247-253", "id_number": "CaltechAUTHORS:FONnp09", "issn": "0028-3908", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:FONnp09", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "PHS", "grant_number": "NS31669" }, { "agency": "PHS", "grant_number": "DA17173" }, { "agency": "PHS", "grant_number": "DA17279" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Philip Morris External Research Fund" }, { "agency": "NARSAD Young Investigator Award" } ] }, "doi": "10.1016/j.neuropharm.2008.08.021", "pmcid": "PMC2645341", "primary_object": { "basename": "nihms87064.pdf", "url": "https://authors.library.caltech.edu/records/tjk3d-0dw59/files/nihms87064.pdf" }, "resource_type": "article", "pub_year": "2008", "author_list": "Fonck, Carlos; Nashmi, Raad; et el." }, { "id": "https://authors.library.caltech.edu/records/m4e9f-2xw36", "eprint_id": 73914, "eprint_status": "archive", "datestamp": "2023-08-19 22:56:00", "lastmod": "2023-10-20 22:54:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Price-K-L", "name": { "family": "Price", "given": "Kerry L." } }, { "id": "Bower-K-S", "name": { "family": "Bower", "given": "Kiowa S." } }, { "id": "Thompson-A-J", "name": { "family": "Thompson", "given": "Andrew J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "Sarah C. R." }, "orcid": "0000-0001-9410-9805" } ] }, "title": "A Hydrogen Bond in Loop A Is Critical for the Binding and Function of the 5-HT_3 Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2008 American Chemical Society. ACS AuthorChoice. \n\nReceived 22 November 2007. Published online 22 May 2008. Published in print 1 June 2008. \n\nWe thank The Wellcome Trust (S.C.R.L. is a Wellcome Trust Senior Research Fellow in Basic Biomedical Science) and the U.S. National Institutes of Health (Grants NS11756 and NS34407) for funding.\n\nPublished - bi800222n.pdf
", "abstract": "The binding sites of Cys-loop receptors are formed from at least six loops (A\u2212F). Here we have used mutagenesis, radioligand binding, voltage clamp electrophysiology, and homology modeling to probe the role of two residues in loop A of the 5-HT_3 receptor: Asn128 and Glu129. The data show that substitution of Asn128, with a range of alternative natural and unnatural amino acids, changed the EC_(50) (from \u223c10-fold more potent to \u223c10-fold less potent than that of the wild type), increased the maximal peak current for mCPBG compared to 5-HT (R_(max)) 2\u221219-fold, and decreased n_H, indicating this residue is involved in receptor gating; we propose Asn128 faces away from the binding pocket and plays a role in facilitating transitions between conformational states. Substitutions of Glu129 resulted in functional receptors only when the residue could accept a hydrogen bond, but with both these and other substitutions, no [^3H]granisetron binding could be detected, indicating a role in ligand binding. We propose that Glu129 faces into the binding pocket, where, through its ability to hydrogen bond, it plays a critical role in ligand binding. Thus, the data support a modified model of the 5-HT_3 receptor binding site and show that loop A plays a critical role in both the ligand binding and function of this receptor.", "date": "2008-06-17", "date_type": "published", "publication": "Biochemistry", "volume": "47", "number": "24", "publisher": "American Chemical Society", "pagerange": "6370-6377", "id_number": "CaltechAUTHORS:20170201-090914461", "issn": "0006-2960", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170201-090914461", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" } ] }, "doi": "10.1021/bi800222n", "pmcid": "PMC264937", "primary_object": { "basename": "bi800222n.pdf", "url": "https://authors.library.caltech.edu/records/m4e9f-2xw36/files/bi800222n.pdf" }, "resource_type": "article", "pub_year": "2008", "author_list": "Price, Kerry L.; Bower, Kiowa S.; et el." }, { "id": "https://authors.library.caltech.edu/records/0hssh-j2290", "eprint_id": 76420, "eprint_status": "archive", "datestamp": "2023-08-19 22:55:26", "lastmod": "2023-10-25 15:59:30", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Maurer-J-A", "name": { "family": "Maurer", "given": "Joshua A." } }, { "id": "Elmore-D-E", "name": { "family": "Elmore", "given": "Donald E." } }, { "id": "Clayton-D", "name": { "family": "Clayton", "given": "Daniel" } }, { "id": "Xiong-Li", "name": { "family": "Xiong", "given": "Li" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Confirming the Revised C-Terminal Domain of the MscL Crystal Structure", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2008 The Biophysical Society. \n\nReceived 6 December 2007, Accepted 23 January 2008, Available online 6 January 2009. \n\nThis work was partially supported by the National Institutes of Health Program Project Grant GM-62532. Additional support was provided by the Office of the Dean at Wellesley College. \n\nJoshua A. Maurer and Donald E. Elmore contributed equally to this work.", "abstract": "The structure of the C-terminal domain of the mechanosensitive channel of large conductance (MscL) has generated significant controversy. As a result, several structures have been proposed for this region: the original crystal structure (1MSL) of the Mycobacterium tuberculosis homolog (Tb), a model of the Escherichia coli homolog, and, most recently, a revised crystal structure of Tb-MscL (2OAR). To understand which of these structures represents a physiological conformation, we measured the impact of mutations to the C-terminal domain on the thermal stability of Tb-MscL using circular dichroism and performed molecular dynamics simulations of the original and the revised crystal structures of Tb-MscL. Our results imply that this region is helical and adopts an \u03b1-helical bundle conformation similar to that observed in the E. coli MscL model and the revised Tb-MscL crystal structure.", "date": "2008-06-15", "date_type": "published", "publication": "Biophysical Journal", "volume": "94", "number": "12", "publisher": "Biophysical Society", "pagerange": "4662-4667", "id_number": "CaltechAUTHORS:20170409-064941478", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170409-064941478", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-62532" }, { "agency": "Wellesley College" } ] }, "doi": "10.1529/biophysj.107.127365", "pmcid": "PMC2397327", "resource_type": "article", "pub_year": "2008", "author_list": "Maurer, Joshua A.; Elmore, Donald E.; et el." }, { "id": "https://authors.library.caltech.edu/records/p1twm-dts78", "eprint_id": 102698, "eprint_status": "archive", "datestamp": "2023-08-22 10:57:42", "lastmod": "2023-10-20 00:25:06", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Shao-Xuesi-M", "name": { "family": "Shao", "given": "Xuesi M." } }, { "id": "Tan-Wenbin", "name": { "family": "Tan", "given": "Wenbin" } }, { "id": "Xiu-Joanne", "name": { "family": "Xiu", "given": "Joanne" } }, { "id": "Puskar-N-L", "name": { "family": "Puskar", "given": "Nyssa" } }, { "id": "Fonck-C", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Feldman-J-L", "name": { "family": "Feldman", "given": "Jack L." } } ] }, "title": "\u03b14* Nicotinic Receptors in preBotzinger Complex Mediate Cholinergic/Nicotinic Modulation of Respiratory Rhythm", "ispublished": "pub", "full_text_status": "public", "keywords": "L9\u2032A knock-in mice; \u03b14 subunit; preB\u00f6tzinger complex; inspiratory neurons; hypoglossal nucleus; nicotine", "note": "\u00a9 2008 Society for Neuroscience. \n\nReceived May 11, 2007; revised Dec. 4, 2007; accepted Dec. 4, 2007. \n\nThis work was supported by Tobacco-Related Disease Research Program (California) Grants 10RT-0241 and 12RT-0245, by National Institutes of Health Grants HL40959 and DA17279, and by Philip Morris USA/International.\n\nPublished - 519.full.pdf
", "abstract": "Acetylcholine and nicotine can modulate respiratory patterns by acting on nicotinic acetylcholine receptors (nAChRs) in the preB\u00f6tzinger complex (preB\u00f6tC). To further explore the molecular composition of these nAChRs, we studied a knock-in mouse strain with a leucine-to-alanine mutation in the M2 pore-lining region (L9\u2032A) of the nAChR \u03b14 subunit; this mutation renders \u03b14-containing receptors hypersensitive to agonists. We recorded respiratory-related rhythmic motor activity from hypoglossal nerve (XIIn) and patch-clamped preB\u00f6tC inspiratory neurons in an in vitro medullary slice preparation from neonatal mice. Nicotine affected respiratory rhythm at concentrations \u223c100-fold lower in the homozygous L9\u2032A knock-in mice compared with wild-type mice. Bath application of 5 nm nicotine increased the excitability of preB\u00f6tC inspiratory neurons, increased respiratory frequency, and induced tonic/seizure-like activities in XIIn in L9\u2032A mice, effects similar to those induced by 1 \u03bcm nicotine in wild-type mice. In L9\u2032A mice, microinjection of low nanomolar concentrations of nicotine into the preB\u00f6tC increased respiratory frequency, whereas injection into the ipsilateral hypoglossal (XII) nucleus induced tonic/seizure-like activity. The \u03b14*-selective nAChR antagonist dihydro-\u03b2-erythroidine produced opposite effects and blocked the nicotinic responses. These data, showing that nAChRs in the preB\u00f6tC and XII nucleus in L9'A mice are hypersensitive to nicotine and endogenous ACh, suggest that functional \u03b14* nAChRs are present in the preB\u00f6tC. They mediate cholinergic/nicotinic modulation of the excitability of preB\u00f6tC inspiratory neurons and of respiratory rhythm. Furthermore, functional \u03b14* nAChRs are present in XII nucleus and mediate cholinergic/nicotinic modulation of tonic activity in XIIn.", "date": "2008-01-09", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "28", "number": "2", "publisher": "Society for Neuroscience", "pagerange": "519-528", "id_number": "CaltechAUTHORS:20200421-112849285", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200421-112849285", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "10RT-0241" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "12RT-0245" }, { "agency": "NIH", "grant_number": "HL40959" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "Philip Morris USA/International" } ] }, "doi": "10.1523/jneurosci.3666-07.2008", "pmcid": "PMC3477875", "primary_object": { "basename": "519.full.pdf", "url": "https://authors.library.caltech.edu/records/p1twm-dts78/files/519.full.pdf" }, "resource_type": "article", "pub_year": "2008", "author_list": "Shao, Xuesi M.; Tan, Wenbin; et el." }, { "id": "https://authors.library.caltech.edu/records/yd462-sr067", "eprint_id": 102709, "eprint_status": "archive", "datestamp": "2023-08-19 21:59:04", "lastmod": "2023-10-20 00:25:38", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Drenan-R-M", "name": { "family": "Drenan", "given": "Ryan M." }, "orcid": "0000-0002-8141-8577" }, { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Imoukhuede-P-I", "name": { "family": "Imoukhuede", "given": "Princess" } }, { "id": "Just-H", "name": { "family": "Just", "given": "Herwig" } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Subcellular Trafficking, Pentameric Assembly, and Subunit Stoichiometry of Neuronal Nicotinic Acetylcholine Receptors Containing Fluorescently Labeled \u03b16 and \u03b23 Subunits", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2008 The American Society for Pharmacology and Experimental Therapeutics. \n\nReceived July 2, 2007; accepted October 11, 2007. \n\nWe thank members of the Lester laboratory for helpful advice and discussion, including Cagdas Son, Rigo Pantoja, and Fraser Moss. Special thanks to Fraser Moss and Monica Liu for help with molecular biology and Bruce Cohen for help with electrophysiology. \n\nThis work was supported by the Plum Foundation; National Institutes of Health (NIH) Grants DA017279, DA019375, DA009121, and NS11756; and by Philip Morris International/USA. R.N. was supported by fellowships from the Elizabeth Ross Foundation, University of California Office of the President Tobacco Related Disease Research Program (UCOP TRDRP) Grant 10FT-0174, and the National Alliance for Research on Schizophrenia and Depression. H.J. was supported by the Austrian Science Fund (Fonds zur F\u00f6rderung der wissenschaftlichen Forschung; Erwin Schr\u00f6dinger Fellowship J2486). R.M.D. was supported by a fellowship from UCOP TRDRP (15FT-0030) and an NIH National Research Service Award (DA021492).", "abstract": "Neuronal nicotinic acetylcholine (ACh) receptors are ligand-gated, cation-selective ion channels. Nicotinic receptors containing \u03b14, \u03b16, \u03b22, and \u03b23 subunits are expressed in midbrain dopaminergic neurons, and they are implicated in the response to smoked nicotine. Here, we have studied the cell biological and biophysical properties of receptors containing \u03b16 and \u03b23 subunits by using fluorescent proteins fused within the M3-M4 intracellular loop. Receptors containing fluorescently tagged \u03b23 subunits were fully functional compared with receptors with untagged \u03b23 subunits. We find that \u03b23- and \u03b16-containing receptors are highly expressed in neurons and that they colocalize with coexpressed, fluorescent \u03b14 and \u03b22 subunits in neuronal soma and dendrites. F\u00f6rster resonance energy transfer (FRET) reveals efficient, specific assembly of \u03b23 and \u03b16 into nicotinic receptor pentamers of various subunit compositions. Using FRET, we demonstrate directly that only a single \u03b23 subunit is incorporated into nicotinic acetylcholine receptors (nAChRs) containing this subunit, whereas multiple subunit stoichiometries exist for \u03b14- and \u03b16-containing receptors. Finally, we demonstrate that nicotinic ACh receptors are localized in distinct microdomains at or near the plasma membrane using total internal reflection fluorescence (TIRF) microscopy. We suggest that neurons contain large, intracellular pools of assembled, functional nicotinic receptors, which may provide them with the ability to rapidly up-regulate nicotinic responses to endogenous ligands such as ACh, or to exogenous agents such as nicotine. Furthermore, this report is the first to directly measure nAChR subunit stoichiometry using FRET and plasma membrane localization of \u03b16- and \u03b23-containing receptors using TIRF.", "date": "2008-01", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "73", "number": "1", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "27-41", "id_number": "CaltechAUTHORS:20200422-074016834", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200422-074016834", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Plum Foundation" }, { "agency": "NIH", "grant_number": "DA017279" }, { "agency": "NIH", "grant_number": "DA019375" }, { "agency": "NIH", "grant_number": "DA009121" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "Philip Morris International" }, { "agency": "Elizabeth Ross Foundation" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "10FT-0174" }, { "agency": "National Alliance for Research on Schizophrenia and Depression" }, { "agency": "Fonds zur F\u00f6rderung der wissenschaftlichen Forschung (FWF)", "grant_number": "J2486" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "15FT-0030" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "DA021492" } ] }, "doi": "10.1124/mol.107.039180", "resource_type": "article", "pub_year": "2008", "author_list": "Drenan, Ryan M.; Nashmi, Raad; et el." }, { "id": "https://authors.library.caltech.edu/records/we2g8-fbv08", "eprint_id": 8982, "eprint_status": "archive", "datestamp": "2023-08-22 10:40:25", "lastmod": "2023-10-16 21:50:53", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Pless-S-A", "name": { "family": "Pless", "given": "Stephan A." } }, { "id": "Dibas-M-I", "name": { "family": "Dibas", "given": "Mohammed I." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Lynch-J-W", "name": { "family": "Lynch", "given": "Joseph W." } } ] }, "title": "Conformational variability of the glycine receptor M2 domain in response to activation by different agonists", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2007 the American Society for Biochemistry and Molecular Biology. \n\nReceived for publication, August 6, 2007 , and in revised form, September 20, 2007. Originally published In Press as doi:10.1074/jbc.M706468200 on October 2, 2007. \n\nThis project was supported by the National Health and Medical Research Council of Australia (NHMRC) and by the US National Institutes of Health (NS-11756). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\"in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. \n\nWe thank Dr. Tim Webb for helpful discussions. \n\n[S.A.P was] [s]upported by an International Postgraduate Research Scholarship from the University of Queensland. \n\n[M.I.D. was] [s]upported by an National Research Service Award from the National Institutes of Health. \n\n[J.W.L. was] [s]upported by a National Health and Medical Research Council of Australia Senior Research Fellowship.\n\nPublished - PLEjbc07.pdf
", "abstract": "Models describing the structural changes mediating cys-loop receptor activation generally give little attention to the possibility that different agonists may promote activation via distinct M2 pore-lining domain structural rearrangements. We investigated this question by comparing the effects of different ligands on the conformation of the external portion of the homomeric \u03b11 glycine receptor M2 domain. Conformational flexibility was assessed by tethering a rhodamine fluorophore to cysteines introduced at the 19' or 22' positions and monitoring fluorescence and current changes during channel activation. During glycine activation, fluorescence of the label attached to R19'C increased by ~20% and the emission peak shifted to lower wavelengths, consistent with a more hydrophobic fluorophore environment. In contrast, ivermectin activated the receptors without producing a fluorescence change. Although taurine and \u03b2-alanine were weak partial agonists at the a1R19'C GlyR, they induced large fluorescence changes. Propofol, which drastically enhanced these currents, did not induce a glycine-like blue-shift in the spectral emission peak. The inhibitors, strychnine and picrotoxin, elicited fluorescence and current changes as expected for a competitive antagonist and an open channel blocker, respectively. Glycine and taurine (or \u03b2-alanine) also produced an increase and a decrease, respectively, in the fluorescence of a label attached to the nearby L22'C residue. Thus, results from two separate labelled residues support the conclusion that the GlyR M2 domain responds with distinct conformational changes to activation by different agonists.", "date": "2007-12-07", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "282", "number": "49", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "36057-36067", "id_number": "CaltechAUTHORS:PLEjbc07", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:PLEjbc07", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Health and Medical Research Council (NHMRC)" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "University of Queensland" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1074/jbc.M706468200", "primary_object": { "basename": "PLEjbc07.pdf", "url": "https://authors.library.caltech.edu/records/we2g8-fbv08/files/PLEjbc07.pdf" }, "resource_type": "article", "pub_year": "2007", "author_list": "Pless, Stephan A.; Dibas, Mohammed I.; et el." }, { "id": "https://authors.library.caltech.edu/records/pwejm-7sb78", "eprint_id": 102763, "eprint_status": "archive", "datestamp": "2023-08-22 10:22:07", "lastmod": "2023-10-20 00:29:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Tapper-A-R", "name": { "family": "Tapper", "given": "Andrew R." } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri L." } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Nicotine responses in hypersensitive and knockout \u03b14 mice account for tolerance to both hypothermia and locomotor suppression in wild-type mice", "ispublished": "pub", "full_text_status": "public", "keywords": "addiction; nicotinic receptors; locomotion", "note": "\u00a9 2007 the American Physiological Society. \n\nReceived 17 March 2007; Accepted 19 August 2007; Published online 1 November 2007; Published in print 1 November 2007. \n\nFunding from National Research Service Award (A. R. Tapper), National Institute of Drug Abuse Grants DA-17279 and DA-19375, California Tobacco-Related Disease Research Program, and Philip Morris USA/International is gratefully acknowledged.\n\nAccepted Version - physiolgenomics.00063.2007_acc.pdf
", "abstract": "Nicotinic receptors containing the \u03b14 subunit (\u03b14* nAChRs) have high sensitivity and are widely expressed in the central nervous system, yet their contributions to behavioral tolerance, a hallmark of nicotine dependence, are unclear. To evaluate the contribution of \u03b14* and non-\u03b14 nAChRs in the development of tolerance to hypothermia and locomotor suppression, \u03b14 knockout (KO), hypersensitive Leu9\u2032Ala \u03b14 knock-in, and wild-type (WT) mice received daily nicotine injections, and their behaviors were compared. Repeated selective activation of \u03b14* nAChRs in Leu9\u2032Ala mice produced profound tolerance to hypothermia over 7 days, whereas no tolerance was observed in \u03b14 KO animals. The summed time course and temperature response (after appropriate normalizations) from these two mutant mouse strains resembled the time course of WT tolerance. In addition, daily selective activation of \u03b14* nAChRs elicited locomotor activation in Leu9\u2032Ala mice, but nicotine suppressed activity in \u03b14 KO mice and this did not change with daily drug exposure. Again, appropriately combined responses from the two mutant strains resembled the biphasic nicotine-induced activity in WT animals. Thus, by analyzing nicotinic responses in two complementary mouse lines, one lacking \u03b14* nAChRs, the other expressing hypersensitive \u03b14* nAChRs, one can accurately separate non-\u03b14 nAChR responses from \u03b14 nAChR responses, and one can also account for WT tolerance to both hypothermia and locomotor suppression. Our study suggests a new paradigm for bridging the gap between genetic manipulation of a single receptor and whole animal behavioral studies and shows that activation of \u03b14* nAChRs is both necessary and sufficient for the expression of tolerance.", "date": "2007-11", "date_type": "published", "publication": "Physiological Genomics", "volume": "31", "number": "3", "publisher": "American Physiological Society", "pagerange": "422-428", "id_number": "CaltechAUTHORS:20200424-071823168", "issn": "1094-8341", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200424-071823168", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH Predoctoral Fellowship" }, { "agency": "NIH", "grant_number": "DA-17279" }, { "agency": "NIH", "grant_number": "DA-19375" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Philip Morris USA/International" } ] }, "doi": "10.1152/physiolgenomics.00063.2007", "primary_object": { "basename": "physiolgenomics.00063.2007_acc.pdf", "url": "https://authors.library.caltech.edu/records/pwejm-7sb78/files/physiolgenomics.00063.2007_acc.pdf" }, "resource_type": "article", "pub_year": "2007", "author_list": "Tapper, Andrew R.; McKinney, Sheri L.; et el." }, { "id": "https://authors.library.caltech.edu/records/8jqxz-csh34", "eprint_id": 102569, "eprint_status": "archive", "datestamp": "2023-08-22 10:09:51", "lastmod": "2023-10-20 00:18:34", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Rodriguez-E-A", "name": { "family": "Rodriguez", "given": "Erik A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 1: Minimizing misacylation", "ispublished": "pub", "full_text_status": "public", "keywords": "nonsense suppression; frameshift suppression; tRNA; opal suppressor tRNA; unnatural amino acid incorporation", "note": "\u00a9 2007 RNA Society. Published by Cold Spring Harbor Laboratory Press. \n\nReceived May 30, 2007. Accepted July 9, 2007. Published in Advance August 13, 2007. \n\nE.A.R. is a National Science Foundation Predoctoral Fellow. This work was supported by the NIH (NS 34407 and NS 11756).\n\nPublished - RNA-2007-Rodriguez-1703-14.pdf
", "abstract": "The incorporation of unnatural amino acids site-specifically is a valuable technique for structure\u2013function studies, incorporation of biophysical probes, and determining protein\u2013protein interactions. THG73 is an amber suppressor tRNA used extensively for the incorporation of >100 different residues in over 20 proteins, but under certain conditions THG73 is aminoacylated in vivo by endogenous aminoacyl-tRNA synthetase. Similar aminoacylation is seen with the Escherichia coli Asn amber suppressor tRNA, which has also been used to incorporate UAAs in many studies. We now find that the natural amino acid placed on THG73 is Gln. Since the E. coli GlnRS recognizes positions in the acceptor stem, we made several acceptor stem mutations in the second to fourth positions on THG73. All mutations reduce aminoacylation in vivo and allow for the selection of highly orthogonal tRNAs. To show the generality of these mutations, we created opal suppressor tRNAs that show less aminoacylation in Xenopus oocytes relative to THG73. We have created a library of Tetrahymena thermophila Gln amber suppressor tRNAs that will be useful for determining optimal suppressor tRNAs for use in other eukaryotic cells.", "date": "2007-10", "date_type": "published", "publication": "RNA", "volume": "13", "number": "10", "publisher": "RNA Society", "pagerange": "1703-1714", "id_number": "CaltechAUTHORS:20200415-144146916", "issn": "1355-8382", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200415-144146916", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NSF Graduate Research Fellowship" }, { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1261/rna.666807", "pmcid": "PMC1986802", "primary_object": { "basename": "RNA-2007-Rodriguez-1703-14.pdf", "url": "https://authors.library.caltech.edu/records/8jqxz-csh34/files/RNA-2007-Rodriguez-1703-14.pdf" }, "resource_type": "article", "pub_year": "2007", "author_list": "Rodriguez, Erik A.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/wxbta-0ym05", "eprint_id": 102570, "eprint_status": "archive", "datestamp": "2023-08-22 10:09:56", "lastmod": "2023-10-20 00:18:36", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Rodriguez-E-A", "name": { "family": "Rodriguez", "given": "Erik A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: Evaluating suppression efficiency", "ispublished": "pub", "full_text_status": "public", "keywords": "nonsense suppression; frameshift suppression; tRNA; opal suppressor tRNA; unnatural amino acid incorporation", "note": "\u00a9 2007 RNA Society. Published by Cold Spring Harbor Laboratory Press. \n\nReceived May 30, 2007. Accepted July 9, 2007. Published in Advance August 13, 2007. \n\nE.A.R. is a National Science Foundation Predoctoral Fellow. This work was supported by the NIH (NS 34407 and NS 11756).\n\nPublished - RNA-2007-Rodriguez-1715-22.pdf
", "abstract": "The incorporation of unnatural amino acids into proteins is a valuable tool for addition of biophysical probes, bio-orthogonal functionalities, and photoreactive cross-linking agents, although these approaches often require quantities of protein that are difficult to access with chemically aminoacylated tRNAs. THG73 is an amber suppressor tRNA that has been used extensively, incorporating over 100 residues in 20 proteins. In vitro studies have shown that the Escherichia coli Asn amber suppressor (ENAS) suppresses better than THG73. However, we report here that ENAS suppresses with <26% of the efficiency of THG73 in Xenopus oocytes. We then tested the newly developed Tetrahymena thermophila Gln amber suppressor (TQAS) tRNA library, which contains mutations in the second to fourth positions of the acceptor stem. The acceptor stem mutations have no adverse effect on suppression efficiency and, in fact, can increase the suppression efficiency. Combining mutations causes an averaging of suppression efficiency, and increased suppression efficiency does not correlate with increased \u0394G of the acceptor stem. We created a T. thermophila opal suppressor, TQOpS\u2032, which shows \u223c50% suppression efficiency relative to THG73. The TQAS tRNA library, composed of functional suppressor tRNAs, has been created and will allow for screening in eukaryotic cells, where rapid analysis of large libraries is not feasible.", "date": "2007-10", "date_type": "published", "publication": "RNA", "volume": "13", "number": "10", "publisher": "RNA Society", "pagerange": "1715-1722", "id_number": "CaltechAUTHORS:20200415-144744313", "issn": "1355-8382", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200415-144744313", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NSF Graduate Research Fellowship" }, { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1261/rna.667607", "pmcid": "PMC1986817", "primary_object": { "basename": "RNA-2007-Rodriguez-1715-22.pdf", "url": "https://authors.library.caltech.edu/records/wxbta-0ym05/files/RNA-2007-Rodriguez-1715-22.pdf" }, "resource_type": "article", "pub_year": "2007", "author_list": "Rodriguez, Erik A.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/xk50b-w6k61", "eprint_id": 102711, "eprint_status": "archive", "datestamp": "2023-08-22 10:02:44", "lastmod": "2023-10-20 00:25:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Teper-Yaroslav", "name": { "family": "Teper", "given": "Yaroslav" } }, { "id": "Whyte-Douglas", "name": { "family": "Whyte", "given": "Douglas" } }, { "id": "Cahir-Elizabeth", "name": { "family": "Cahir", "given": "Elizabeth" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Grady-Sharon-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Marks-Michael-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce N." }, "orcid": "0000-0003-2913-6238" }, { "id": "Fonck-Carlos", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "McClure-Begley-Tristan-D", "name": { "family": "McClure-Begley", "given": "Tristan" } }, { "id": "McIntosh-J-Michael", "name": { "family": "McIntosh", "given": "J. Michael" } }, { "id": "Labarca-Cesar-G", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Lawrence-Andrew", "name": { "family": "Lawrence", "given": "Andrew" } }, { "id": "Chen-Feng", "name": { "family": "Chen", "given": "Feng" } }, { "id": "Gantois-Ilse", "name": { "family": "Gantois", "given": "Ilse" } }, { "id": "Davies-Philip-J", "name": { "family": "Davies", "given": "Philip J." } }, { "id": "Petrou-Steven", "name": { "family": "Petrou", "given": "Steven" } }, { "id": "Murphy-Mark", "name": { "family": "Murphy", "given": "Mark" } }, { "id": "Waddington-John", "name": { "family": "Waddington", "given": "John" } }, { "id": "Horne-Malcolm-K", "name": { "family": "Horne", "given": "Malcolm K." } }, { "id": "Berkovic-Samuel-F", "name": { "family": "Berkovic", "given": "Samuel F." } }, { "id": "Drago-John", "name": { "family": "Drago", "given": "John" } } ] }, "title": "Nicotine-Induced Dystonic Arousal Complex in a Mouse Line Harboring a Human Autosomal-Dominant Nocturnal Frontal Lobe Epilepsy Mutation", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotine; epilepsy; ADNFLE; dystonia; mecamylamine; synaptosome", "note": "\u00a9 2007 Society for Neuroscience. \n\nReceived Nov. 21, 2006; revised Aug. 1, 2007; accepted Aug. 1, 2007. \n\nThis work was supported by National Health and Medical Research Council (NHMRC) Program Grant 236805 and National Institutes of Health Grant MH53631 to J.M.M.; National Institute on Drug Abuse Grants DA003194, DA012242, and DA015663; National Institute of Neurological Disorders and Stroke Grants NS43800 and NS046464; and Science Foundation Ireland Grant 02-1N1B227. J.D. is an NHMRC Practitioner fellow. We thank Vincenzo Ferreri, Jim Massalas, Keith Buxton, and Yvette Wilson for technical assistance and Dr. Rachael Nally and Associate Prof. Ian Gordon for advice with statistical analysis. We are grateful to Dr. Jim Boulter for providing genomic DNA clones used to build the original L9S' targeting construct. We are grateful to Prof. Ingrid Scheffer for critical input and discussion on clinical aspects of ADNFLE.\n\nPublished - 10128.full.pdf
Supplemental Material - Human_ADNFLE_video.mpg
Supplemental Material - S248F_DAC1_video.mpg
Supplemental Material - S248F_DAC2.mpg
", "abstract": "We generated a mouse line harboring an autosomal-dominant nocturnal frontal lobe epilepsy (ADNFLE) mutation: the \u03b14 nicotinic receptor S248F knock-in strain. In this mouse, modest nicotine doses (1\u20132 mg/kg) elicit a novel behavior termed the dystonic arousal complex (DAC). The DAC includes stereotypical head movements, body jerking, and forelimb dystonia; these behaviors resemble some core features of ADNFLE. A marked Straub tail is an additional component of the DAC. Similar to attacks in ADNFLE, the DAC can be partially suppressed by the sodium channel blocker carbamazepine or by pre-exposure to a very low dose of nicotine (0.1 mg/kg). The DAC is centrally mediated, genetically highly penetrant, and, surprisingly, not associated with overt ictal electrical activity as assessed by (1) epidural or frontal lobe depth-electrode electroencephalography or (2) hippocampal c-fos-regulated gene expression. Heterozygous knock-in mice are partially protected from nicotine-induced seizures. The noncompetitive antagonist mecamylamine does not suppress the DAC, although it suppresses high-dose nicotine-induced wild-type-like seizures. Experiments on agonist-induced \u2078\u2076Rb\u207a and neurotransmitter efflux from synaptosomes and on \u03b14S248F\u03b22 receptors expressed in oocytes confirm that the S248F mutation confers resistance to mecamylamine blockade. Genetic background, gender, and mutant gene expression levels modulate expression of the DAC phenotype in mice. The S248F mouse thus appears to provide a model for the paroxysmal dystonic element of ADNFLE semiology. Our model complements what is seen in other ADNFLE animal models. Together, these mice cover the spectrum of behavioral and electrographic events seen in the human condition.", "date": "2007-09-19", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "27", "number": "38", "publisher": "Society for Neuroscience", "pagerange": "10128-10142", "id_number": "CaltechAUTHORS:20200422-080631596", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200422-080631596", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Health and Medical Research Council (NHMRC)", "grant_number": "236805" }, { "agency": "NIH", "grant_number": "MH53631" }, { "agency": "NIH", "grant_number": "DA003194" }, { "agency": "NIH", "grant_number": "DA012242" }, { "agency": "NIH", "grant_number": "DA015663" }, { "agency": "NIH", "grant_number": "NS43800" }, { "agency": "NIH", "grant_number": "NS046464" }, { "agency": "Science Foundation, Ireland", "grant_number": "02-1N1B227" } ] }, "doi": "10.1523/jneurosci.3042-07.2007", "pmcid": "PMC6672658", "primary_object": { "basename": "S248F_DAC2.mpg", "url": "https://authors.library.caltech.edu/records/xk50b-w6k61/files/S248F_DAC2.mpg" }, "related_objects": [ { "basename": "10128.full.pdf", "url": "https://authors.library.caltech.edu/records/xk50b-w6k61/files/10128.full.pdf" }, { "basename": "Human_ADNFLE_video.mpg", "url": "https://authors.library.caltech.edu/records/xk50b-w6k61/files/Human_ADNFLE_video.mpg" }, { "basename": "S248F_DAC1_video.mpg", "url": "https://authors.library.caltech.edu/records/xk50b-w6k61/files/S248F_DAC1_video.mpg" } ], "resource_type": "article", "pub_year": "2007", "author_list": "Teper, Yaroslav; Whyte, Douglas; et el." }, { "id": "https://authors.library.caltech.edu/records/snvda-46319", "eprint_id": 8655, "eprint_status": "archive", "datestamp": "2023-08-22 09:58:42", "lastmod": "2023-10-16 21:37:54", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Wang-Jinti", "name": { "family": "Wang", "given": "Jinti" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Establishing an Ion Pair Interaction in the Homomeric {rho}1 {gamma}-Aminobutyric Acid Type A Receptor That Contributes to the Gating Pathway", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2007 the American Society for Biochemistry and Molecular Biology. \n\nReceived for publication, March 16, 2007, and in revised form, May 30, 2007. Originally published In Press as doi:10.1074/jbc.M702314200 on July 2, 2007 \n\nWe thank Michael Torrice for the preparation of Nha. \n\nThis work was supported by National Institutes of Health Grants NS 34407 and NS 11756. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. \n\nThe on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1 and S2.\n\nPublished - WANjbc07.pdf
Supplemental Material - WANjbc07supp.pdf
", "abstract": "{gamma}-Aminobutyric acid type A (GABAA) receptors are members of the Cys-loop superfamily of ligand-gated ion channels. Upon agonist binding, the receptor undergoes a structural transition from the closed to the open state, but the mechanism of gating is not well understood. Here we utilized a combination of conventional mutagenesis and the high precision methodology of unnatural amino acid incorporation to study the gating interface of the human homopentameric {rho}1 GABAA receptor. We have identified an ion pair interaction between two conserved charged residues, Glu92 in loop 2 of the extracellular domain and Arg258 in the pre-M1 region. We hypothesize that the salt bridge exists in the closed state by kinetic measurements and free energy analysis. Several other charged residues at the gating interface are not critical to receptor function, supporting previous conclusions that it is the global charge pattern of the gating interface that controls receptor function in the Cys-loop superfamily.", "date": "2007-09-07", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "282", "number": "36", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "26210-26216", "id_number": "CaltechAUTHORS:WANjbc07", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:WANjbc07", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1074/jbc.M702314200", "primary_object": { "basename": "WANjbc07.pdf", "url": "https://authors.library.caltech.edu/records/snvda-46319/files/WANjbc07.pdf" }, "related_objects": [ { "basename": "WANjbc07supp.pdf", "url": "https://authors.library.caltech.edu/records/snvda-46319/files/WANjbc07supp.pdf" } ], "resource_type": "article", "pub_year": "2007", "author_list": "Wang, Jinti; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/1h2t6-76680", "eprint_id": 8527, "eprint_status": "archive", "datestamp": "2023-08-22 09:43:18", "lastmod": "2023-10-16 21:32:09", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Jankowsky-J-L", "name": { "family": "Jankowsky", "given": "Joanna L." } }, { "id": "Younkin-L-H", "name": { "family": "Younkin", "given": "Linda H." } }, { "id": "Gonzales-V", "name": { "family": "Gonzales", "given": "Victoria" } }, { "id": "Fadale-D-J", "name": { "family": "Fadale", "given": "Daniel J." } }, { "id": "Slunt-H-H", "name": { "family": "Slunt", "given": "Hilda H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Younkin-S-G", "name": { "family": "Younkin", "given": "Steven G." } }, { "id": "Borchelt-D-R", "name": { "family": "Borchelt", "given": "David R." } } ] }, "title": "Rodent A\u03b2 Modulates the Solubility and Distribution of Amyloid Deposits in Transgenic Mice", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2007 the American Society for Biochemistry and Molecular Biology. \n\nReceived for publication, November 30, 2006, and in revised form, April 20, 2007. Originally published In Press as doi:10.1074/jbc.M611050200 on June 15, 2007. Originally published In Press as doi:10.1074/jbc.M611050200 on June 7, 2007. \n\nWe thank Gay Rudow for advice on stereology, Eduardo Marcora for help with the Odyssey imager, and Neil Segil for sharing his ultracentrifuge for this work. We are grateful to Takeda Chemical Industries Co., Ltd. for providing antibodies BAN50, BNT77, BA27, and BC05, and to Konrad Beyreuther and Andreas Weidemann for sharing the 22C11 antibody. We thank David Teplow for providing mouse and human Abeta peptide standards, and David Holtzman for sharing peptide electrophoresis protocols. \n\nThis work was supported in part by NIA, National Institutes of Health Grants 1 P50 AG-14-248 and 1 P01 AG-98-003 (to D. R. B.) and AG-06-656 (to S. G. Y.), the Robert H. and Clarice Smith and Abigail Van Buren Alzheimer's Disease Research Program (to S. G. Y.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.\n\nPublished - JANjbc07.pdf
", "abstract": "The amino acid sequence of amyloid precursor protein (APP) is highly conserved, and age-related Abeta aggregates have been described in a variety of vertebrate animals, with the notable exception of mice and rats. Three amino acid substitutions distinguish mouse and human Abeta that might contribute to their differing properties in vivo. To examine the amyloidogenic potential of mouse Abeta, we studied several lines of transgenic mice overexpressing wild-type mouse amyloid precursor protein (moAPP) either alone or in conjunction with mutant PS1 (PS1dE9). Neither overexpression of moAPP alone nor co-expression with PS1dE9 caused mice to develop Alzheimer-type amyloid pathology by 24 months of age. We further tested whether mouse Abeta could accelerate the deposition of human Abeta by crossing the moAPP transgenic mice to a bigenic line expressing human APPswe with PS1dE9. The triple transgenic animals (moAPP x APPswe/PS1dE9) produced 20% more Abeta but formed amyloid deposits no faster and to no greater extent than APPswe/PS1dE9 siblings. Instead, the additional mouse Abeta increased the detergent solubility of accumulated amyloid and exacerbated amyloid deposition in the vasculature. These findings suggest that, although mouse Abeta does not influence the rate of amyloid formation, the incorporation of Abeta peptides with differing sequences alters the solubility and localization of the resulting aggregates.", "date": "2007-08-03", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "282", "number": "31", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "22707-22720", "id_number": "CaltechAUTHORS:JANjbc07", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:JANjbc07", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "1 P50 AG-14-248" }, { "agency": "NIH", "grant_number": "1 P01 AG-98-003" }, { "agency": "NIH", "grant_number": "AG-06-656" }, { "agency": "Robert H. and Clarice Smith and Abigail Van Buren Alzheimer's Disease Research Program" } ] }, "doi": "10.1074/jbc.M611050200", "pmcid": "PMC4435736", "primary_object": { "basename": "JANjbc07.pdf", "url": "https://authors.library.caltech.edu/records/1h2t6-76680/files/JANjbc07.pdf" }, "resource_type": "article", "pub_year": "2007", "author_list": "Jankowsky, Joanna L.; Younkin, Linda H.; et el." }, { "id": "https://authors.library.caltech.edu/records/q8dwj-r4s47", "eprint_id": 98422, "eprint_status": "archive", "datestamp": "2023-08-22 09:43:02", "lastmod": "2023-10-18 17:21:00", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nashmi-Raad", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Deshpande-Purnima", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "McKinney-Sheri-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Grady-Sharon-R", "name": { "family": "Grady", "given": "Sharon R." } }, { "id": "Whiteaker-Paul", "name": { "family": "Whiteaker", "given": "Paul" } }, { "id": "Huang-Qi", "name": { "family": "Huang", "given": "Qi" } }, { "id": "McClure-Begley-Tristan", "name": { "family": "McClure-Begley", "given": "Tristan" } }, { "id": "Lindstrom-Jon-M", "name": { "family": "Lindstrom", "given": "Jon M." } }, { "id": "Labarca-Cesar-G", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Collins-Allan-C", "name": { "family": "Collins", "given": "Allan C." } }, { "id": "Marks-Michael-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Chronic Nicotine Cell Specifically Upregulates Functional \u03b14* Nicotinic Receptors: Basis for Both Tolerance in Midbrain and Enhanced Long-Term Potentiation in Perforant Path", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotine addiction; GFP; knock-in mouse; nicotinic receptor; GABAergic; dopaminergic", "note": "\u00a9 2007 Society for Neuroscience. \n\nReceived Dec. 14, 2006; revised June 12, 2007; accepted June 14, 2007. \n\nThis work was supported by National Institutes of Health Grants DA-3194, DA-15663, DA-19655 (at Boulder); DA-09121, DA-17279, NS-11756 (at Caltech); DA-19375 (at both Boulder and Caltech); and NS-11323 (at the University of Pennsylvania); by the California Tobacco-Related Disease Research Project (Grant 12RT-0245); by Philip Morris USA/International at Caltech; by the Elizabeth Ross Foundation and National Alliance for Research on Schizophrenia and Depression (postdoctoral fellowships to R.N.); and by the W. M. Keck and Plum Foundations. We thank J. Drago for \u03b14 knock-out mice. We thank V. Vargas, J. Wang, J. Silva, and S. Pease for excellent mouse husbandry; D. A. Dougherty, E. M. Schuman, J. L. Jankowsky, and J. Schwartz for comments on this manuscript; and B. N. Cohen, R. M. Drenan, C. Fonck, J. Miwa, R. Pantoja, and A. Tapper for discussions.\n\nPublished - 8202.full.pdf
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", "abstract": "Understanding effects of chronic nicotine requires identifying the neurons and synapses whose responses to nicotine itself, and to endogenous acetylcholine, are altered by continued exposure to the drug. To address this problem, we developed mice whose \u03b14 nicotinic receptor subunits are replaced by normally functioning fluorescently tagged subunits, providing quantitative studies of receptor regulation at micrometer resolution. Chronic nicotine increased \u03b14 fluorescence in several regions; among these, midbrain and hippocampus were assessed functionally. Although the midbrain dopaminergic system dominates reward pathways, chronic nicotine does not change \u03b14* receptor levels in dopaminergic neurons of ventral tegmental area (VTA) or substantia nigra pars compacta. Instead, upregulated, functional \u03b14* receptors localize to the GABAergic neurons of the VTA and substantia nigra pars reticulata. In consequence, GABAergic neurons from chronically nicotine-treated mice have a higher basal firing rate and respond more strongly to nicotine; because of the resulting increased inhibition, dopaminergic neurons have lower basal firing and decreased response to nicotine. In hippocampus, chronic exposure to nicotine also increases \u03b14* fluorescence on glutamatergic axons of the medial perforant path. In hippocampal slices from chronically treated animals, acute exposure to nicotine during tetanic stimuli enhances induction of long-term potentiation in the medial perforant path, showing that the upregulated \u03b14* receptors in this pathway are also functional. The pattern of cell-specific upregulation of functional \u03b14* receptors therefore provides a possible explanation for two effects of chronic nicotine: sensitization of synaptic transmission in forebrain and tolerance of dopaminergic neuron firing in midbrain.", "date": "2007-08-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "27", "number": "31", "publisher": "Society for Neuroscience", "pagerange": "8202-8218", "id_number": "CaltechAUTHORS:20190905-080227761", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190905-080227761", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-3194" }, { "agency": "NIH", "grant_number": "DA-15663" }, { "agency": "NIH", "grant_number": "DA-19655" }, { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "DA-17279" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "DA-19375" }, { "agency": "NIH", "grant_number": "NS-11323" }, { "agency": "California Tobacco-Related Disease Research Project", "grant_number": "12RT-0245" }, { "agency": "Philip Morris USA" }, { "agency": "Elizabeth Ross Foundation" }, { "agency": "National Alliance for Research on Schizophrenia and Depression" }, { "agency": "W. M. Keck Foundation" }, { "agency": "Plum Foundation" } ] }, "doi": "10.1523/jneurosci.2199-07.2007", "pmcid": "PMC6673074", "primary_object": { "basename": "FigS13.gif", "url": "https://authors.library.caltech.edu/records/q8dwj-r4s47/files/FigS13.gif" }, "related_objects": [ { "basename": "FigS14a.gif", "url": "https://authors.library.caltech.edu/records/q8dwj-r4s47/files/FigS14a.gif" }, { "basename": "FigS9.gif", "url": "https://authors.library.caltech.edu/records/q8dwj-r4s47/files/FigS9.gif" }, { "basename": "Supplemental_Legend.pdf", "url": "https://authors.library.caltech.edu/records/q8dwj-r4s47/files/Supplemental_Legend.pdf" }, { "basename": "8202.full.pdf", "url": "https://authors.library.caltech.edu/records/q8dwj-r4s47/files/8202.full.pdf" }, { "basename": "FigS10.gif", "url": "https://authors.library.caltech.edu/records/q8dwj-r4s47/files/FigS10.gif" }, { "basename": "FigS11.gif", "url": "https://authors.library.caltech.edu/records/q8dwj-r4s47/files/FigS11.gif" }, { "basename": "FigS12.gif", "url": "https://authors.library.caltech.edu/records/q8dwj-r4s47/files/FigS12.gif" } ], "resource_type": "article", "pub_year": "2007", "author_list": "Nashmi, Raad; Xiao, Cheng; et el." }, { "id": "https://authors.library.caltech.edu/records/pcfph-dj488", "eprint_id": 102785, "eprint_status": "archive", "datestamp": "2023-08-19 20:10:30", "lastmod": "2023-10-20 00:30:12", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Damaj-M-I", "name": { "family": "Damaj", "given": "M. I." } }, { "id": "Fonck-C", "name": { "family": "Fonck", "given": "C." } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "M. J." } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "P." } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "C." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Collins-A-C", "name": { "family": "Collins", "given": "A. C." } }, { "id": "Martin-B-R", "name": { "family": "Martin", "given": "B. R." } } ] }, "title": "Genetic Approaches Identify Differential Roles for \u03b1\u2084\u03b2\u2082* Nicotinic Receptors in Acute Models of Antinociception in Mice", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2007 by The American Society for Pharmacology and Experimental Therapeutics. \n\nReceived August 17, 2006; accepted March 16, 2007. \n\nThis work was supported by National Institute on Drug Abuse Grants DA-12610, DA-0527, DA-03194, and DA-17279, by the National Institute of Mental Health (MH-49176), and by the California Tobacco-Related Disease Research Program.\n\nPublished - 1161.full.pdf
", "abstract": "The effects of nicotine on the tail-flick and hot-plate tests were determined to identify nicotinic receptor subtypes responsible for spinally and supraspinally mediated nicotine analgesia in knockin mice expressing hypersensitive \u03b14 nicotinic receptors (L9\u2032S), in seven inbred mouse strains (C57BL/6, DBA/2, A/2, CBA/2, BALB/cByJ, C3H/HeJ, and 129/SvEv), and in two F1 hybrids (B6CBAF1 and B6D2F1). L9\u2032S heterozygotes were \u223c6-fold more sensitive to the antinociceptive effects of nicotine than the wild-type controls in the hot-plate test but not in the tail-flick assay. Large differences in the effects of nicotine were also observed with both tests for the seven mouse strains. A/J and 129 mice were 6- to 8-fold more sensitive than CBA and BALB mice. In addition, B6CBAF1 hybrid mice were even less sensitive than CBA mice. Nicotinic binding sites were measured in three spinal cord regions and the hindbrain of the inbred strains. Significant differences in cytisine-sensitive, high affinity [\u00b9\u00b2\u2075I]epibatidine binding site levels (\u03b1\u2082\u03b2\u2082* subtypes), but not in \u00b9\u00b2\u2075I-\u03b1-bungarotoxin binding (\u03b17* subtypes), were observed. Significant negative correlations between cytisine-sensitive [\u00b9\u00b2\u2075I]epibatidine binding and nicotine ED50 for both tests were noted. Our results indicate that \u03b1\u2084\u03b2\u2082* acetylcholine nicotinic receptors (nAChR) are important in mediating nicotine analgesia in supraspinal responses, while also showing that \u03b1\u2084\u03b2\u2082*-nAChR and at least one other nAChR subtype appear to modulate spinal actions.", "date": "2007-05-01", "date_type": "published", "publication": "Journal of Pharmacology and Experimental Therapeutics", "volume": "321", "number": "3", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "1161-1169", "id_number": "CaltechAUTHORS:20200427-083757686", "issn": "0022-3565", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-083757686", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-12610" }, { "agency": "NIH", "grant_number": "DA-0527" }, { "agency": "NIH", "grant_number": "DA-03194" }, { "agency": "NIH", "grant_number": "DA-17279" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "California Tobacco-Related Disease Research Program" } ] }, "doi": "10.1124/jpet.106.112649", "primary_object": { "basename": "1161.full.pdf", "url": "https://authors.library.caltech.edu/records/pcfph-dj488/files/1161.full.pdf" }, "resource_type": "article", "pub_year": "2007", "author_list": "Damaj, M. I.; Fonck, C.; et el." }, { "id": "https://authors.library.caltech.edu/records/amvr7-dmz63", "eprint_id": 55543, "eprint_status": "archive", "datestamp": "2023-08-19 20:00:43", "lastmod": "2023-10-20 22:22:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lerchner-W", "name": { "family": "Lerchner", "given": "Walter" } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Slimko-E-M", "name": { "family": "Slimko", "given": "Eric M." } }, { "id": "van-Trigt-L", "name": { "family": "van Trigt", "given": "Laurent" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Anderson-D-J", "name": { "family": "Anderson", "given": "David J." }, "orcid": "0000-0001-6175-3872" } ] }, "title": "Reversible Silencing of Neuronal Excitability in Behaving Mice by a Genetically Targeted, Ivermectin-Gated Cl^\u2212 Channel", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2007 Elsevier Inc. Under an Elsevier user license. \n\nReceived: November 20, 2006; Revised: February 9, 2007; Accepted: February 22, 2007; Published: April 4, 2007. \n\nWe thank Christof Koch and Herwig Just for helpful discussions, Gabriele Mosconi for laboratory management, Shirley Pease and Janet Baer, D.V.M. for mouse management and assistance with surgical procedures, Dirk Neumann for help with statistics, and Steven Smith for help with the Ethovision Software. D.J.A. is an Investigator of the Howard Hughes Medical Institute. Support for this work was provided by the Moore foundation Conscious Mouse Project and by NIH grant MH-67867 to H.A.L.\n\nSupplemental Material - mmc1.pdf
Supplemental Material - mmc2.mpeg
", "abstract": "Several genetic strategies for inhibiting neuronal function in mice have been described, but no system that directly suppresses membrane excitability and is triggered by a systemically administered drug, has been validated in awake behaving animals. We expressed unilaterally in mouse striatum a modified heteromeric ivermectin (IVM)-gated chloride channel from C. elegans (GluCl\u03b1\u03b2), systemically administered IVM, and then assessed amphetamine-induced rotational behavior. Rotation was observed as early as 4 hr after a single intraperitoneal IVM injection (10 mg/kg), reached maximal levels by 12 hr, and was almost fully reversed by 4 days. Multiple cycles of silencing and recovery could be performed in a single animal. In striatal slice preparations from GluCl\u03b1\u03b2-expressing animals, IVM rapidly suppressed spiking. The two-subunit GluCl/IVM system permits \"intersectional\" strategies designed to increase the cellular specificity of silencing in transgenic animals.", "date": "2007-04-05", "date_type": "published", "publication": "Neuron", "volume": "54", "number": "1", "publisher": "Elsevier", "pagerange": "35-49", "id_number": "CaltechAUTHORS:20150305-114442340", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150305-114442340", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "Gordon and Betty Moore Foundation" }, { "agency": "NIH", "grant_number": "MH-67867" } ] }, "doi": "10.1016/j.neuron.2007.02.030", "primary_object": { "basename": "mmc1.pdf", "url": "https://authors.library.caltech.edu/records/amvr7-dmz63/files/mmc1.pdf" }, "related_objects": [ { "basename": "mmc2.mpeg", "url": "https://authors.library.caltech.edu/records/amvr7-dmz63/files/mmc2.mpeg" } ], "resource_type": "article", "pub_year": "2007", "author_list": "Lerchner, Walter; Xiao, Cheng; et el." }, { "id": "https://authors.library.caltech.edu/records/sgjy5-pmn32", "eprint_id": 102589, "eprint_status": "archive", "datestamp": "2023-08-22 08:11:57", "lastmod": "2023-10-20 00:19:23", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Padgett-C-L", "name": { "family": "Padgett", "given": "Claire L." } }, { "id": "Hanek-A-P", "name": { "family": "Hanek", "given": "Ariele P." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "Sarah C. R." }, "orcid": "0000-0001-9410-9805" } ] }, "title": "Unnatural Amino Acid Mutagenesis of the GABA_A Receptor Binding Site Residues Reveals a Novel Cation-\u03c0 Interaction between GABA and \u03b2\u2082Tyr97", "ispublished": "pub", "full_text_status": "public", "keywords": "ligand-gated ion channel; Cys-loop receptor; cation\u2013\u03c0 interaction; GABA_A receptor binding site; unnatural amino acids; homology model", "note": "\u00a9 2007 Society for Neuroscience.\n\nReceived July 28, 2006; revised Dec. 14, 2006; accepted Dec. 14, 2006.\n\nThis work was supported by the Wellcome Trust (S.C.R.L. is a Wellcome Trust Senior Research Fellow in Basic Biomedical Science), Merck Sharpe and Dohme, the Biotechnology and Biological Sciences Research Council (a CASE studentship to C.L.P.), and the National Institutes of Health (NIH) (NS11756; NS34407). A.P.H. is a recipient of NIH Predoctoral Trainee Grant 5T32GMO7616.\n\nPublished - 886.full.pdf
Supplemental Material - Fig1S.pdf
", "abstract": "The binding pockets of Cys-loop receptors are dominated by aromatic amino acids. In the GABA_A receptor \u03b1\u2081Phe65, \u03b2\u2082Tyr97, \u03b2\u2082Tyr157, and \u03b2\u2082Tyr205 are present at the \u03b2\u2082/\u03b1\u2081 interface and have been implicated in forming an important part of the GABA binding site. Here, we have probed interactions of these residues using subtle chemical changes: unnatural amino acid mutagenesis was used to introduce a range of Phe analogs, and mutant receptors expressed in oocytes were studied using voltage-clamp electrophysiology. Serial mutations at \u03b2\u208297 revealed a \u223c20-fold increase in EC\u2085\u2080 with the addition of each fluorine atom to a phenylalanine, indicating a cation\u2013\u03c0 interaction between GABA and this residue. This is the first example of a cation\u2013\u03c0 interaction in loop A of a Cys-loop receptor. Along with previous studies that identified cation\u2013\u03c0 interactions in loop B and loop C, the result emphasizes that the location of this interaction is not conserved in the Cys-loop family. The data further show that \u03b1\u208165 (in loop D) is tolerant to subtle changes. Conversely, mutating either \u03b2\u2082Tyr157 (in loop B) or \u03b2\u2082Tyr205 (in loop C) to Phe substantially disrupts receptor function. Substitution of 4-F-Phe, however, at either position, or 4-MeO-Phe at \u03b2\u2082Tyr157, resulted in receptors with wild-type EC\u2085\u2080 values, suggesting a possible hydrogen bond. The molecular scale insights provided by these data allow the construction of a model for GABA docking to the agonist binding site of the GABA_A receptor.", "date": "2007-01-24", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "27", "number": "4", "publisher": "Society for Neuroscience", "pagerange": "886-892", "id_number": "CaltechAUTHORS:20200416-150305764", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200416-150305764", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "Merck Sharp & Dohme Research Laboratories" }, { "agency": "Dohme" }, { "agency": "Biotechnology and Biological Sciences Research Council (BBSRC)" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "5T32GMO7616" } ] }, "doi": "10.1523/jneurosci.4791-06.2007", "pmcid": "PMC2649369", "primary_object": { "basename": "886.full.pdf", "url": "https://authors.library.caltech.edu/records/sgjy5-pmn32/files/886.full.pdf" }, "related_objects": [ { "basename": "Fig1S.pdf", "url": "https://authors.library.caltech.edu/records/sgjy5-pmn32/files/Fig1S.pdf" } ], "resource_type": "article", "pub_year": "2007", "author_list": "Padgett, Claire L.; Hanek, Ariele P.; et el." }, { "id": "https://authors.library.caltech.edu/records/m6y1q-h0w88", "eprint_id": 73873, "eprint_status": "archive", "datestamp": "2023-08-19 19:34:22", "lastmod": "2023-10-24 21:03:55", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Cashin-A-L", "name": { "family": "Cashin", "given": "Amanda L." } }, { "id": "Torrice-M-M", "name": { "family": "Torrice", "given": "Michael M." } }, { "id": "McMenimen-K-A", "name": { "family": "McMenimen", "given": "Kathryn A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Chemical-Scale Studies on the Role of a Conserved Aspartate in Preorganizing the Agonist Binding Site of the Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2007 American Chemical Society. \n\nReceived 11 August 2006. Published online 30 December 2006. Published in print 1 January 2007. \n\nThis work was supported by the National Institutes of Health (Grants NS 34407 and NS 11756) and by the George Hoag Family Foundation. \n\nWe thank Tingwei Mu for providing the unnatural amino acid Akp.\n\nAccepted Version - nihms61839.pdf
", "abstract": "The nicotinic acetylcholine receptor and related Cys-loop receptors are ligand-gated ion channels that mediate fast synaptic transmission throughout the central and peripheral nervous system. A highly conserved aspartate residue (D89) that is near the agonist binding site but does not directly contact the ligand plays a critical part in receptor function. Here we probe the role of D89 using unnatural amino acid mutagenesis coupled with electrophysiology. Homology modeling implicates several hydrogen bonds involving D89. We find that no single hydrogen bond is essential to proper receptor function. Apparently, the side chain of D89 establishes a redundant network of hydrogen bonds; these bonds preorganize the agonist binding site by positioning a critical tryptophan residue that directly contacts the ligand. Earlier studies of the D89N mutant led to the proposal that a negative charge at this position is essential for receptor function. However, we find that receptors with neutral side chains at position 89 can function well, if the side chain is less perturbing than the amide of asparagine (nitro or keto groups allow function) or if a compensating backbone mutation is introduced to relieve unfavorable electrostatics.", "date": "2007-01-23", "date_type": "published", "publication": "Biochemistry", "volume": "46", "number": "3", "publisher": "American Chemical Society", "pagerange": "630-639", "id_number": "CaltechAUTHORS:20170131-102250754", "issn": "0006-2960", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170131-102250754", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "George Hoag Family Foundation" } ] }, "doi": "10.1021/bi061638b", "pmcid": "PMC3164877", "primary_object": { "basename": "nihms61839.pdf", "url": "https://authors.library.caltech.edu/records/m6y1q-h0w88/files/nihms61839.pdf" }, "resource_type": "article", "pub_year": "2007", "author_list": "Cashin, Amanda L.; Torrice, Michael M.; et el." }, { "id": "https://authors.library.caltech.edu/records/kaaxn-ckq62", "eprint_id": 6628, "eprint_status": "archive", "datestamp": "2023-08-22 07:39:41", "lastmod": "2023-10-16 20:26:36", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Ahern-C-A", "name": { "family": "Ahern", "given": "Christopher A." } }, { "id": "Eastwood-A-L", "name": { "family": "Eastwood", "given": "Amy L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Horn-Richard", "name": { "family": "Horn", "given": "Richard" } } ] }, "title": "A Cation\u2013\u03c0 Interaction between Extracellular TEA and an Aromatic Residue in Potassium Channels", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2006 The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nSubmitted: 21 August 2006. Accepted: 27 October 2006. Published online Nov 27 2006. doi:10.1085/jgp.200609654 \n\nWe thank Benoit Roux for extensive discussions and advice about Poisson-Boltzmann calculations, Carol Deutsch for comments on the manuscript, Adrian Gross for the gift of a TEAs-Cl sample, and Stephanie Huang, Purnima Deshpande, and Lindsey Ingleby for help with the oocytes. \n\nThis research was sponsored by grants from the National Institutes of Health (GM079427, NS11756, and NS34407). \n\nLawrence G. Palmer served as editor.\n\nPublished - AHEjgp06.pdf
", "abstract": "Open-channel blockers such as tetraethylammonium (TEA) have a long history as probes of the permeation pathway of ion channels. High affinity blockade by extracellular TEA requires the presence of an aromatic amino acid at a position that sits at the external entrance of the permeation pathway (residue 449 in the eukaryotic voltage-gated potassium channel Shaker). We investigated whether a cation\u2013{pi} interaction between TEA and such an aromatic residue contributes to TEA block using the in vivo nonsense suppression method to incorporate a series of increasingly fluorinated Phe side chains at position 449. Fluorination, which is known to decrease the cation\u2013{pi} binding ability of an aromatic ring, progressively increased the inhibitory constant Ki for the TEA block of Shaker. A larger increase in Ki was observed when the benzene ring of Phe449 was substituted by nonaromatic cyclohexane. These results support a strong cation\u2013{pi} component to the TEA block. The data provide an empirical basis for choosing between Shaker models that are based on two classes of reported crystal structures for the bacterial channel KcsA, showing residue Tyr82 in orientations either compatible or incompatible with a cation\u2013{pi} mechanism. We propose that the aromatic residue at this position in Shaker is favorably oriented for a cation\u2013{pi} interaction with the permeation pathway. This choice is supported by high level ab initio calculations of the predicted effects of Phe modifications on TEA binding energy.", "date": "2006-12", "date_type": "published", "publication": "Journal of General Physiology", "volume": "128", "number": "6", "publisher": "Rockefeller University Press", "pagerange": "649-657", "id_number": "CaltechAUTHORS:AHEjgp06", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:AHEjgp06", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM079427" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" } ] }, "doi": "10.1085/jgp.200609654", "pmcid": "PMC2151593", "primary_object": { "basename": "AHEjgp06.pdf", "url": "https://authors.library.caltech.edu/records/kaaxn-ckq62/files/AHEjgp06.pdf" }, "resource_type": "article", "pub_year": "2006", "author_list": "Ahern, Christopher A.; Eastwood, Amy L.; et el." }, { "id": "https://authors.library.caltech.edu/records/5a3cf-98860", "eprint_id": 102796, "eprint_status": "archive", "datestamp": "2023-08-22 06:29:42", "lastmod": "2023-10-20 00:30:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Miwa-J-M", "name": { "family": "Miwa", "given": "Julie M." } }, { "id": "Stevens-T-R", "name": { "family": "Stevens", "given": "Tanya R." } }, { "id": "King-S-L", "name": { "family": "King", "given": "Sarah L." } }, { "id": "Caldarone-B-J", "name": { "family": "Caldarone", "given": "Barbara J." } }, { "id": "Ibanez-Tallon-I", "name": { "family": "Ibanez-Tallon", "given": "Ines" } }, { "id": "Xiao-Cheng", "name": { "family": "Xiao", "given": "Cheng" }, "orcid": "0000-0001-9649-7450" }, { "id": "Fitzsimonds-R-M", "name": { "family": "Fitzsimonds", "given": "Reiko Maki" } }, { "id": "Pavlides-C", "name": { "family": "Pavlides", "given": "Constantine" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Picciotto-M-R", "name": { "family": "Picciotto", "given": "Marina R." } }, { "id": "Heintz-N", "name": { "family": "Heintz", "given": "Nathaniel" } } ] }, "title": "The Prototoxin lynx1 Acts on Nicotinic Acetylcholine Receptors to Balance Neuronal Activity and Survival In Vivo", "ispublished": "pub", "full_text_status": "public", "keywords": "MOLNEURO; SYSNEURO", "note": "\u00a9 2006 Elsevier Inc. \n\nReceived 21 December 2004, Revised 21 October 2005, Accepted 19 July 2006, Available online 6 September 2006. \n\nWe wish to thank A.L. Beaudet for generously providing the \u03b17 and \u03b22 nAChR mutant mice used in these experiments. Special thanks to Dr. Andreas Walz, Dr. S. Selesnick, and Dr. P. Gutin. For helpful scientific discussions and/or technical support, thanks to C. Fletcher, R.A. Lester, A.B. Tekinay, R. Nashmi, A. Tapper, R. Pantoja, F. Moss, and B. Cohen and other members of HAL lab; B. Switzer of NSA, A. Kolar, S. Krueger, and A. Aslantas. Support for M.R.P., S.L.K., B.J.C., and T.R.S.: DA00436, DA15241, and AA15632; for J.M.M., NIH CA09673; for N.H., J.M.M., and I.I.-T., HHMI and NIH NIH R21 NS047751; for H.A.L. and C.X., DA-17279 and CA Tobacco-Related Disease Research Project.\n\nSupplemental Material - 1-s2.0-S0896627306005940-mmc1.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) affect a wide array of biological processes, including learning and memory, attention, and addiction. lynx1, the founding member of a family of mammalian prototoxins, modulates nAChR function in vitro by altering agonist sensitivity and desensitization kinetics. Here we demonstrate, through the generation of lynx1 null mutant mice, that lynx1 modulates nAChR signaling in vivo. Its loss decreases the EC\u2085\u2080 for nicotine by \u223c10-fold, decreases receptor desensitization, elevates intracellular calcium levels in response to nicotine, and enhances synaptic efficacy. lynx1 null mutant mice exhibit enhanced performance in specific tests of learning and memory. Consistent with reports that mutations resulting in hyperactivation of nAChRs can lead to neurodegeneration, aging lynx1 null mutant mice exhibit a vacuolating degeneration that is exacerbated by nicotine and ameliorated by null mutations in nAChRs. We conclude that lynx1 functions as an allosteric modulator of nAChR function in vivo, balancing neuronal activity and survival in the CNS.", "date": "2006-09-07", "date_type": "published", "publication": "Neuron", "volume": "51", "number": "5", "publisher": "Cell Press", "pagerange": "587-600", "id_number": "CaltechAUTHORS:20200427-095213669", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-095213669", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA00436" }, { "agency": "NIH", "grant_number": "DA15241" }, { "agency": "NIH", "grant_number": "AA15632" }, { "agency": "NIH", "grant_number": "CA09673" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "NIH", "grant_number": "R21 NS047751" }, { "agency": "NIH", "grant_number": "DA-17279" }, { "agency": "California Tobacco-Related Disease Research Program" } ] }, "doi": "10.1016/j.neuron.2006.07.025", "primary_object": { "basename": "1-s2.0-S0896627306005940-mmc1.pdf", "url": "https://authors.library.caltech.edu/records/5a3cf-98860/files/1-s2.0-S0896627306005940-mmc1.pdf" }, "resource_type": "article", "pub_year": "2006", "author_list": "Miwa, Julie M.; Stevens, Tanya R.; et el." }, { "id": "https://authors.library.caltech.edu/records/dq13n-4bt76", "eprint_id": 5967, "eprint_status": "archive", "datestamp": "2023-08-22 05:52:36", "lastmod": "2023-10-16 20:00:08", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Rodriguez-E-A", "name": { "family": "Rodriguez", "given": "Erik A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "In vivo incorporation of multiple unnatural amino acids through nonsense and frameshift suppression", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotinic receptor; tRNA; quadruplet codon; stop codon; protein engineering", "note": "\u00a9 2006 by the National Academy of Sciences \n\nEdited by Peter G. Schultz, The Scripps Research Institute, La Jolla, CA, and approved April 21, 2006 (received for review December 15, 2005). Published online before print May 25, 2006, 10.1073/pnas.0510817103 \n\nE.A.R. is a National Science Foundation Predoctoral Fellow. This work was supported by National Institutes of Health Grants NS-34407 and NS-11756. \n\nAuthor contributions: E.A.R. designed research; E.A.R. performed research; E.A.R., H.A.L., and D.A.D. analyzed data; and E.A.R., H.A.L., and D.A.D. wrote the paper. \n\nConflict of interest statement: No conflicts declared. \n\nThis paper was submitted directly (Track II) to the PNAS office.\n\nPublished - RODpnas06.pdf
", "abstract": "Site-specific incorporation of unnatural amino acids (UAAs) into proteins is a valuable tool for studying structure\u2013function relationships, incorporating biophysical probes, and elucidating protein\u2013protein interactions. In higher eukaryotic cells, the methodology is currently limited to incorporation of a single UAA in response to a stop codon, which is known as nonsense suppression. Frameshift suppression is a unique methodology for incorporating UAAs in response to quadruplet codons, but currently, it is mostly limited to in vitro protein translation systems. Here, we evaluate the viability of frameshift suppression in Xenopus oocytes. We demonstrate UAA incorporation by using yeast phenylalanine frameshift suppressor (YFFS) tRNAs that recognize two different quadruplet codons (CGGG and GGGU) in vivo. Suppression efficiency of the YFFS tRNAs increases nonlinearly with the amount of injected tRNA, suggesting a significant competition with endogenous, triplet-recognizing tRNA. Both frameshift suppressor tRNAs are less efficient than the amber suppressor tRNA THG73 (Tetrahymena thermophila G73), which has been used extensively for UAA incorporation in Xenopus oocytes. However, the two YFFS tRNAs are more \"orthogonal\" to the Xenopus system than THG73, and they offer a viable replacement when suppressing at promiscuous sites. To illustrate the potential of combining nonsense and frameshift suppression, we have site-specifically incorporated two and three UAAs simultaneously into a neuroreceptor expressed in vivo.", "date": "2006-06-06", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "103", "number": "23", "publisher": "National Academy of Sciences", "pagerange": "8650-8655", "id_number": "CaltechAUTHORS:RODpnas06", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:RODpnas06", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NSF Predoctoral Fellowship" }, { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1073/pnas.0510817103", "pmcid": "PMC1482635", "primary_object": { "basename": "RODpnas06.pdf", "url": "https://authors.library.caltech.edu/records/dq13n-4bt76/files/RODpnas06.pdf" }, "resource_type": "article", "pub_year": "2006", "author_list": "Rodriguez, Erik A.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/an46q-pgf85", "eprint_id": 75885, "eprint_status": "archive", "datestamp": "2023-08-22 05:38:12", "lastmod": "2023-10-25 15:23:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Schwarz-J", "name": { "family": "Schwarz", "given": "Johannes" } }, { "id": "Schwarz-S-C", "name": { "family": "Schwarz", "given": "Sigrid C." } }, { "id": "Dorigo-O", "name": { "family": "Dorigo", "given": "Oliver" } }, { "id": "St\u00fctzer-A", "name": { "family": "St\u00fctzer", "given": "Alexandra" } }, { "id": "Wegner-F", "name": { "family": "Wegner", "given": "Florian" } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Gil-J-S", "name": { "family": "Gil", "given": "Jose S." } }, { "id": "Berk-A-J", "name": { "family": "Berk", "given": "Arnold J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Enhanced expression of hypersensitive \u03b14* nAChR in adult mice increases the loss of midbrain dopaminergic neurons", "ispublished": "pub", "full_text_status": "restricted", "keywords": "nicotinic receptor; substantia nigra; cholinergic excitotoxicity; knock-in mouse; helper-dependent adenovirus; Parkinson's disease", "note": "\u00a9 2006 Federation of American Societies for Experimental Biology. \n\nReceived for publication November 28, 2005. Accepted for publication December 19, 2005. \n\nAcknowledgments: Deutsche Forschungsgemeinschaft 704/3\u20131, IZKF-Leipzig TP C27, California; Tobacco-Related Disease Research Program, National Parkinson Foundation, Keck Foundation, Plum Foundation, NIH (NS-11756, MH-49176); and a UCLA Human Gene Therapy seed grant. O.D. was supported by a grant from the American Association of Obstetricians and Gynecologists Foundation (AAOGF). J.S.G. received partial support from U.S. Public Health Service National Research Service award GM07185 and a Cota Robles Fellowship. We thank Ralf Schober for assistance with ICC images.", "abstract": "We describe an inducible genetic model for degeneration of midbrain dopaminergic neurons in adults. In previous studies, knock-in mice expressing hypersensitive M2 domain Leu9'Ser (L9'S) \u03b14 nicotinic receptors (nAChR) at near-normal levels displayed dominant neonatal lethality and dopaminergic deficits in embryonic midbrain, because the hypersensitive nAChR is excitotoxic. However, heterozygous L9'S mice that retain the neomycin resistance cassette (neo) in a neighboring intron express low levels of the mutant allele (\u223c25% of normal levels), and these neo-intact mice are therefore viable and fertile. The neo cassette is flanked by loxP sites. In adult animals, we locally injected helper-dependent adenovirus (HDA) expressing cre recombinase. Local excision of the neo cassette, via cre-mediated recombination, was verified by genomic analysis. In L9'S HDA-cre injected animals, locomotion was reduced both under baseline conditions and after amphetamine application. There was no effect in L9'S HDA-control treated animals or in wild-type (WT) littermates injected with either virus. Immunocytochemical analyses revealed marked losses (> 70%) of dopaminergic neurons in L9'S HDA-cre injected mice compared to controls. At 20\u201333 days postinjection in control animals, the coexpressed marker gene, yellow fluorescent protein (YFP), was expressed in many neurons and few glial cells near the injection, emphasizing the neurotropic utility of the HDA. Thus, HDA-mediated gene transfer into adult midbrain induced sufficient functional expression of cre in dopaminergic neurons to allow for postnatal deletion of neo. This produced increased L9'S mutant nAChR expression, which in turn led to nicotinic cholinergic excitotoxicity in dopaminergic neurons.", "date": "2006-05", "date_type": "published", "publication": "FASEB Journal", "volume": "20", "number": "7", "publisher": "Federation of American Societies for Experimental Biology", "pagerange": "935-946", "id_number": "CaltechAUTHORS:20170408-135451181", "issn": "0892-6638", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170408-135451181", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Deutsche Forschungsgemeinschaft (DFG)", "grant_number": "704/3\u20131" }, { "agency": "ZKF-Leipzig", "grant_number": "TP C27" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "National Parkinson Foundation" }, { "agency": "W. M. Keck Foundation" }, { "agency": "Plum Foundation" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "UCLA Human Gene Therapy seed grant" }, { "agency": "American Association of Obstetricians and Gynecologists Foundation" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM07185" }, { "agency": "Cota Robles Fellowship" } ] }, "doi": "10.1096/fj.05-5497com", "resource_type": "article", "pub_year": "2006", "author_list": "Schwarz, Johannes; Schwarz, Sigrid C.; et el." }, { "id": "https://authors.library.caltech.edu/records/gze51-10s84", "eprint_id": 73944, "eprint_status": "archive", "datestamp": "2023-08-19 17:46:17", "lastmod": "2023-10-24 21:08:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "McMenimen-K-A", "name": { "family": "McMenimen", "given": "Kathryn A." } }, { "id": "Petersson-E-J", "name": { "family": "Petersson", "given": "E. James" }, "orcid": "0000-0003-3854-9210" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Probing the Mg^(2+) Blockade Site of an N-Methyl-d-aspartate (NMDA) Receptor with Unnatural Amino Acid Mutagenesis", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2006 American Chemical Society. \n\nReceived 1 March 2006. Date accepted 21 April 2006. Published online 12 May 2006. Published in print 1 May 2006. \n\nWe thank K. Williams for the NMDA receptor subunit clones and C. Waters for use of the Beckman Institute Biological Imaging Center. This work was supported by grants from the National Institutes of Health (NS-34407, and NS-11756).\n\nSupplemental Material - cb6000944si20060512_000000.doc
", "abstract": "The N-methyl-d-aspartate (NMDA) receptor plays a central role in learning and memory in the mammalian CNS. At normal neuronal resting membrane potentials, the pore of this glutamate-gated ion channel is blocked by a Mg^(2+) ion. Previous work suggests that the Mg^(2+) binding site is quite novel, involving several asparagine residues and a cation\u2013\u03c0 interaction between Mg^(2+) and a conserved tryptophan in the pore. Using unnatural amino acid mutagenesis, we show that no such cation\u2013\u03c0 interaction exists. The implicated tryptophan instead appears to play a structural role that can only be fulfilled by a rigid, flat, hydrophobic residue. This is the first demonstration of unnatural amino acid incorporation in the NMDA receptor, and it opens the way for future investigations of this pivotal neuroreceptor.", "date": "2006-05", "date_type": "published", "publication": "ACS Chemical Biology", "volume": "1", "number": "4", "publisher": "American Chemical Society", "pagerange": "227-234", "id_number": "CaltechAUTHORS:20170201-140122486", "issn": "1554-8929", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170201-140122486", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1021/cb6000944", "primary_object": { "basename": "cb6000944si20060512_000000.doc", "url": "https://authors.library.caltech.edu/records/gze51-10s84/files/cb6000944si20060512_000000.doc" }, "resource_type": "article", "pub_year": "2006", "author_list": "McMenimen, Kathryn A.; Petersson, E. James; et el." }, { "id": "https://authors.library.caltech.edu/records/kn6ta-qhx02", "eprint_id": 102805, "eprint_status": "archive", "datestamp": "2023-08-19 17:38:43", "lastmod": "2023-10-20 00:31:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Sokolova-I-V", "name": { "family": "Sokolova", "given": "Irina V." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Postsynaptic Mechanisms Are Essential for Forskolin-Induced Potentiation of Synaptic Transmission", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2006 by the American Physiological Society. \n\nReceived 13 June 2005; Accepted 22 December 2005; Published online 1 April 2006; Published in print 1 April 2006. \n\nWe are thankful to S. McKinney for neuronal cultures, A. Kleschevnikov for discussion and valuable suggestions, and K. Diba for help in preparing the manuscript. \n\nThis research was supported by National Institute of Mental Health Grant MH-49176. \n\nThe costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.\n\nAccepted Version - jn.00617.2005_acc.pdf
", "abstract": "It has been demonstrated that stimulation of protein kinase A (PKA) results in enhanced synaptic transmission in the hippocampus and other brain areas. To investigate mechanisms of the PKA-mediated potentiation of synaptic transmission, we used rat hippocampal embryonic cultures. In low-density cultures, paired recordings under the perforated patch demonstrated that 15-min forskolin treatment produced long-lasting potentiation of evoked excitatory postsynaptic currents (eEPSCs) mediated by the cAMP/PKA pathway. eEPSC amplitudes increased to 240 \u00b1 10% of baseline after 15 min of forskolin treatment (early). After forskolin washout, eEPSCs declined to a potentiated level. Potentiation was sustained for \u226585 min after forskolin washout and, 60 min after forskolin washout, constituted 152 \u00b1 7% of baseline (late potentiation). Disruption of presynaptic processes with the whole cell configuration and internal solution containing PKA inhibitor peptide did not affect forskolin-induced potentiation. Disruption of postsynaptic processes, in contrast, impaired early potentiation and abolished late potentiation. Study of mEPSCs confirmed the contribution of postsynaptic mechanisms. Forskolin-induced enhancement of mEPSC frequency observed under the perforated patch was attenuated by the whole cell configuration. Forskolin also induced an increase of mEPSC amplitudes in the perforated patch, but not in the whole cell, experiments. Potentiation of eEPSCs was not activity dependent, persisting in the absence of stimulation. NMDA receptor blockade did not abolish forskolin-induced potentiation. In summary, we demonstrate that forskolin-induced potentiation of eEPSCs was mediated by postsynaptic mechanisms, presumably by upregulation of AMPA receptors by phosphorylation.", "date": "2006-04", "date_type": "published", "publication": "Journal of Neurophysiology", "volume": "95", "number": "4", "publisher": "American Physiological Society", "pagerange": "2570-2579", "id_number": "CaltechAUTHORS:20200427-123125281", "issn": "0022-3077", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-123125281", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "MH-49176" } ] }, "doi": "10.1152/jn.00617.2005", "primary_object": { "basename": "jn.00617.2005_acc.pdf", "url": "https://authors.library.caltech.edu/records/kn6ta-qhx02/files/jn.00617.2005_acc.pdf" }, "resource_type": "article", "pub_year": "2006", "author_list": "Sokolova, Irina V.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/r2gfw-ftq42", "eprint_id": 102814, "eprint_status": "archive", "datestamp": "2023-08-22 05:07:17", "lastmod": "2023-10-20 00:31:29", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "CNS Localization of Neuronal Nicotinic Receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "Nicotine; Ventral Tegmental Area; Nicotinic Receptor; Molecular Neuroscience Volume; nAChR Subunit", "note": "\u00a9 2006 Humana Press Inc. \n\nThis work was supported by the California Tobacco-Related Disease Research Program (TRDRP; 12RT-0245), National Institutes of Health grants (NS-11756, DA-17279), and the Phillip Morris External Research Program. R. N. was supported by a postdoctoral fellowship from the TRDRP(10FT-0174) and the Elizabeth Ross Fellowship.\n\nPublished - Nashmi-Lester2006_Article_CNSLocalizationOfNeuronalNicot.pdf
", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are members of the Cys-loop superfamily of pentameric ligand-gated ion channels, which include GABA (A and C), serotonin, and glycine receptors. Currently, 12 neuronal nAChR subunits have been identified (\u03b12\u201310 and \u03b22\u20134) and are generally grouped into \u03b1 subunits, which contain two adjacent cysteine residues essential for ACh binding, and \u03b1 subunits, which lack these residues. The majority of neuronal nAChRs fall into two categories: those that bind agonist with high affinity (nMconcentrations); and those that bind with lower affinity (\u00b5M concentrations). The low-affinity receptors are presumably homomeric \u03b17 receptors that are \u03b1-bungarotoxin sensitive, whereas \u03b14\u03b22 nAChRs account for >90% of the high-affinity nicotinic receptors in the brain (Whiting and Lindstrom, 1986). Their physiological contributions to neurotransmission, signaling, and behavior are not completely understood. Precise mapping of subcellular and neuroanatomical localizations of neuronal nAChR subunits will help elucidate the physiological role of neuronal nAChRs and their role in nicotine addiction.", "date": "2006-02", "date_type": "published", "publication": "Journal of Molecular Neuroscience", "volume": "30", "number": "1-2", "publisher": "Humana Press", "pagerange": "181-184", "id_number": "CaltechAUTHORS:20200427-134509446", "issn": "0895-8696", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-134509446", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "12RT-0245" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "DA-17279" }, { "agency": "Phillip Morris External Research Program" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "10FT-0174" }, { "agency": "Elizabeth Ross Fellowship" } ] }, "doi": "10.1385/jmn:30:1:181", "primary_object": { "basename": "Nashmi-Lester2006_Article_CNSLocalizationOfNeuronalNicot.pdf", "url": "https://authors.library.caltech.edu/records/r2gfw-ftq42/files/Nashmi-Lester2006_Article_CNSLocalizationOfNeuronalNicot.pdf" }, "resource_type": "article", "pub_year": "2006", "author_list": "Nashmi, Raad and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/v7frr-swv57", "eprint_id": 22095, "eprint_status": "archive", "datestamp": "2023-08-19 17:16:09", "lastmod": "2023-10-23 15:40:56", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "CNS Localization of Neuronal Nicotinic Receptors", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2006 Humana Press Inc.\n\nThis work was supported by the California\nTobacco-Related Disease Research Program (TRDRP;\n12RT-0245), National Institutes of Health grants (NS-11756, DA-17279), and the Phillip Morris External\nResearch Program. R. N. was supported by a postdoctoral\nfellowship from the TRDRP (10FT-0174) and\nthe Elizabeth Ross Fellowship.", "abstract": "Nicotinic acetylcholine receptors (nAChRs) are\nmembers of the Cys-loop superfamily of pentameric\nligand-gated ion channels, which include GABA (A\nand C), serotonin, and glycine receptors. Currently, 12\nneuronal nAChR subunits have been identified (", "date": "2006-02", "date_type": "published", "publication": "Journal of Molecular Neuroscience", "volume": "30", "number": "1-2", "publisher": "Humana Press", "pagerange": "181-184", "id_number": "CaltechAUTHORS:20110209-113037940", "issn": "0895-8696", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20110209-113037940", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "12RT-0245" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "DA-17279" }, { "agency": "Phillip Morris External Research Program" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "10FT-0174" }, { "agency": "Elizabeth Ross Fellowship" } ] }, "doi": "10.1385/JMN:30:1:181", "resource_type": "article", "pub_year": "2006", "author_list": "Nashmi, Raad and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/tyjkx-b2x70", "eprint_id": 3329, "eprint_status": "archive", "datestamp": "2023-08-22 04:41:13", "lastmod": "2023-10-16 15:48:25", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Xiu-Xinan", "name": { "family": "Xiu", "given": "Xinan" } }, { "id": "Hanek-A-P", "name": { "family": "Hanek", "given": "Ariele P." } }, { "id": "Wang-Jinti", "name": { "family": "Wang", "given": "Jinti" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "A Unified View of the Role of Electrostatic Interactions in Modulating the Gating of Cys Loop Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2005 the American Society for Biochemistry and Molecular Biology. \n\nReceived for publication, August 5, 2005, and in revised form, October 6, 2005. Originally published In Press as doi:10.1074/jbc.M508635200 on October 10, 2005. \n\nThis work was supported by the National Institutes of Health Grants NS-34407 and NS-11756. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. \n\nThe on-line version of this article (available at http://www.jbc.org) contains Figs. i-v and Table 1.\n\nPublished - XIUjbc05.pdf
", "abstract": "In the Cys loop superfamily of ligand-gated ion channels, a global conformational change, initiated by agonist binding, results in channel opening and the passage of ions across the cell membrane. The detailed mechanism of channel gating is a subject that has lent itself to both structural and electrophysiological studies. Here we defined a gating interface that incorporates elements from the ligand binding domain and transmembrane domain previously reported as integral to proper channel gating. An overall analysis of charged residues within the gating interface across the entire superfamily showed a conserved charging pattern, although no specific interacting ion pairs were conserved. We utilized a combination of conventional mutagenesis and the high precision methodology of unnatural amino acid incorporation to study extensively the gating interface of the mouse muscle nicotinic acetylcholine receptor. We found that charge reversal, charge neutralization, and charge introduction at the gating interface are often well tolerated. Furthermore, based on our data and a reexamination of previously reported data on {gamma}-aminobutyric acid, type A, and glycine receptors, we concluded that the overall charging pattern of the gating interface, and not any specific pairwise electrostatic interactions, controls the gating process in the Cys loop superfamily.", "date": "2005-12-16", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "280", "number": "50", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "41655-41666", "id_number": "CaltechAUTHORS:XIUjbc05", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:XIUjbc05", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1074/jbc.M508635200", "primary_object": { "basename": "XIUjbc05.pdf", "url": "https://authors.library.caltech.edu/records/tyjkx-b2x70/files/XIUjbc05.pdf" }, "resource_type": "article", "pub_year": "2005", "author_list": "Xiu, Xinan; Hanek, Ariele P.; et el." }, { "id": "https://authors.library.caltech.edu/records/b0mw2-cwe70", "eprint_id": 102816, "eprint_status": "archive", "datestamp": "2023-08-22 04:35:37", "lastmod": "2023-10-20 00:31:31", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Fonck-Carlos", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Nashmi-Raad", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Whiteaker-Paul", "name": { "family": "Whiteaker", "given": "Paul" } }, { "id": "Wagenaar-D-A", "name": { "family": "Wagenaar", "given": "Daniel A." }, "orcid": "0000-0002-6222-761X" }, { "id": "Rodrigues-Pinguet-Nivalda-O", "name": { "family": "Rodrigues-Pinguet", "given": "Nivalda" } }, { "id": "Deshpande-Purnima", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "McKinney-Sheri", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Kwoh-Steven", "name": { "family": "Kwoh", "given": "Steven" } }, { "id": "Munoz-Jose", "name": { "family": "Munoz", "given": "Jose" } }, { "id": "Labarca-C-G", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Collins-Allan-C", "name": { "family": "Collins", "given": "Allan C." } }, { "id": "Marks-Michael-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Novel Seizure Phenotype and Sleep Disruptions in Knock-In Mice with Hypersensitive \u03b14* Nicotinic Receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "epilepsy; ADNFLE; nicotinic receptors; sleep disorders; \u03b14\u03b22; nicotine", "note": "\u00a9 2005 Society for Neuroscience. \n\nReceived Feb. 9, 2005; revised Oct. 18, 2005; accepted Oct. 21, 2005. \n\nThis work was supported by National Institute of Health Grants NS46464, NS43800, NS11756, DA10156, DA03194, and DA17279 and a National Institutes of Health\u2013National Research Service Award (C.F.). The California Tobacco-Related Disease Research Program and Philip Morris USA/International are gratefully acknowledged. We thank J. Stitzel for the mouse \u03b14 cDNA, Chana Simon for oocyte data analysis, Robert Paz for help with photography, and Sharon Grady, Andrew Tapper, and Johannes Schwarz for insightful discussion.\n\nPublished - 11396.full.pdf
", "abstract": "A leucine to alanine substitution (L9\u2032A) was introduced in the M2 region of the mouse \u03b14 neuronal nicotinic acetylcholine receptor (nAChR) subunit. Expressed in Xenopus oocytes, \u03b14(L9\u2032A)\u03b22 nAChRs were \u226530-fold more sensitive than wild type (WT) to both ACh and nicotine. We generated knock-in mice with the L9\u2032A mutation and studied their cellular responses, seizure phenotype, and sleep-wake cycle. Seizure studies on \u03b14-mutated animals are relevant to epilepsy research because all known mutations linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) occur in the M2 region of \u03b14or \u03b22 subunits. Thalamic cultures and synaptosomes from L9\u2032A mice were hypersensitive to nicotine-induced ion flux. L9\u2032A mice were \u223c15-fold more sensitive to seizures elicited by nicotine injection than their WT littermates. Seizures in L9\u2032A mice differed qualitatively from those in WT: L9\u2032A seizures started earlier, were prevented by nicotine pretreatment, lacked EEG spike-wave discharges, and consisted of fast repetitive movements. Nicotine-induced seizures in L9\u2032A mice were partial, whereas WT seizures were generalized. When L9\u2032A homozygous mice received a 10 mg/kg nicotine injection, there was temporal and phenomenological separation of mutant and WT-like seizures: an initial seizure \u223c20 s after injection was clonic and showed no EEG changes. A second seizure began 3-4 min after injection, was tonic-clonic, and had EEG spike-wave activity. No spontaneous seizures were detected in L9\u2032A mice during chronic video/EEG recordings, but their sleep-wake cycle was altered. Our findings show that hypersensitive \u03b14* nicotinic receptors in mice mediate changes in the sleep-wake cycle and nicotine-induced seizures resembling ADNFLE.", "date": "2005-12-07", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "25", "number": "49", "publisher": "Society for Neuroscience", "pagerange": "11396-11411", "id_number": "CaltechAUTHORS:20200427-144703784", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-144703784", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS46464" }, { "agency": "NIH", "grant_number": "NS43800" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "DA10156" }, { "agency": "NIH", "grant_number": "DA03194" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Philip Morris USA/International" } ] }, "doi": "10.1523/jneurosci.3597-05.2005", "pmcid": "PMC6725918", "primary_object": { "basename": "11396.full.pdf", "url": "https://authors.library.caltech.edu/records/b0mw2-cwe70/files/11396.full.pdf" }, "resource_type": "article", "pub_year": "2005", "author_list": "Fonck, Carlos; Cohen, Bruce N.; et el." }, { "id": "https://authors.library.caltech.edu/records/5pmf7-4rg66", "eprint_id": 984, "eprint_status": "archive", "datestamp": "2023-08-22 04:29:37", "lastmod": "2023-10-13 22:04:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Jankowsky-J-L", "name": { "family": "Jankowsky", "given": "Joanna L." } }, { "id": "Slunt-H-H", "name": { "family": "Slunt", "given": "Hilda H." } }, { "id": "Gonzales-V", "name": { "family": "Gonzales", "given": "Victoria" } }, { "id": "Savonenko-A-V", "name": { "family": "Savonenko", "given": "Alena V." } }, { "id": "Wen-Jason-C", "name": { "family": "Wen", "given": "Jason C." } }, { "id": "Jenkins-N-A", "name": { "family": "Jenkins", "given": "Nancy A." } }, { "id": "Copeland-N-G", "name": { "family": "Copeland", "given": "Neal G." } }, { "id": "Younkin-L-H", "name": { "family": "Younkin", "given": "Linda H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Younkin-S-G", "name": { "family": "Younkin", "given": "Steven G." } }, { "id": "Borchelt-D-R", "name": { "family": "Borchelt", "given": "David R." } } ] }, "title": "Persistent amyloidosis following suppression of A\u03b2 production in a transgenic model of Alzheimer disease", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2005 Jankowsky et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. \n\nReceived: June 14, 2005; Accepted: August 22, 2005; Published: November 15, 2005. Academic Editor: Adriano Aguzzi, Z\u00fcrich University, Switzerland \n\nWe thank Patrick Tremblay for helpful advice on the tet system at a critical time in the project, and Mark Mayford for sharing the CaMKII\u03b1-tTA mice through Jackson Laboratory. We also thank Fraser Moss for saving several immunoblots with last-minute shipments, Andy Groves for many thoughtful discussions, Neil Segil for generously sharing his laboratory and equipment, Beth Olson, Natasha Bouey, and Yolanda Jackson for outstanding animal care, Debbie Swing for expert microinjection, and Dave Fromholt for genotyping and dissection. We gratefully acknowledge Takeda Chemical Industries for providing antibodies BAN50, BA27, and BC05, Konrad Beyreuther and Andreas Weidemann for providing 22C11 antibody, and Ed Koo for sharing CT15 antibody. This work was supported by grants from the Johns Hopkins Alzheimer's Disease Research Center (JLJ), the National Alliance for Research on Schizophrenia and Depression (Young Investigator Award [JLJ]), the Rose Hills Foundation (JLJ), the Alzheimer's Association (Zenith Award [DRB]), the National Institute of Aging (K01 AG26144\u201301 [JLJ], P50 AGO5146\u201320 [DRB], R01 AG006656\u201316 [SGY], and P01 AG015453 [SGY]), the National Institute of Neurologic Disease and Stoke (R01 NS 047225 [DRB]), and the National Cancer Institute (NAJ and NGC). The funding agencies generously provided for research supplies, animal care, and salary support; the funders of this work had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. \n\nCompeting Interests: The authors have declared that no competing interests exist.\n\nAuthor Contributions: JLJ and DRB designed the study and wrote the manuscript, JLJ, HHS, VG, JCW, LHY, SGY and DRB performed experiments, NAJ and NGC generated transgenic founders, HAL assisted with data interpretation, and AVS performed statistical analyses.\n\nPublished - JANplosmed05.pdf
Supplemental Material - journal.pmed.0020355.sg001.TIF
Supplemental Material - journal.pmed.0020355.sg002.PSD
Supplemental Material - journal.pmed.0020355.sg003.PSD
Supplemental Material - journal.pmed.0020355.sg004.JPG
Supplemental Material - journal.pmed.0020355.sg005.TIF
Supplemental Material - journal.pmed.0020355.sg006.PSD
", "abstract": "Background: The proteases (secretases) that cleave amyloid-\u03b2 (A\u03b2) peptide from the amyloid precursor protein (APP) have been the focus of considerable investigation in the development of treatments for Alzheimer disease. The prediction has been that reducing A\u03b2 production in the brain, even after the onset of clinical symptoms and the development of associated pathology, will facilitate the repair of damaged tissue and removal of amyloid lesions. However, no long-term studies using animal models of amyloid pathology have yet been performed to test this hypothesis. \n\nMethods and Findings: We have generated a transgenic mouse model that genetically mimics the arrest of A\u03b2 production expected from treatment with secretase inhibitors. These mice overexpress mutant APP from a vector that can be regulated by doxycycline. Under normal conditions, high-level expression of APP quickly induces fulminant amyloid pathology. We show that doxycycline administration inhibits transgenic APP expression by greater than 95% and reduces A\u03b2 production to levels found in nontransgenic mice. Suppression of transgenic A\u03b2 synthesis in this model abruptly halts the progression of amyloid pathology. However, formation and disaggregation of amyloid deposits appear to be in disequilibrium as the plaques require far longer to disperse than to assemble. Mice in which APP synthesis was suppressed for as long as 6 mo after the formation of A\u03b2 deposits retain a considerable amyloid load, with little sign of active clearance. \n\nConclusion: This study demonstrates that amyloid lesions in transgenic mice are highly stable structures in vivo that are slow to disaggregate. Our findings suggest that arresting A\u03b2 production in patients with Alzheimer disease should halt the progression of pathology, but that early treatment may be imperative, as it appears that amyloid deposits, once formed, will require additional intervention to clear.", "date": "2005-12", "date_type": "published", "publication": "PLoS Medicine", "volume": "2", "number": "12", "publisher": "Public Library of Science", "pagerange": "Art. No. e355", "id_number": "CaltechAUTHORS:JANplosmed05.886", "issn": "1549-1676", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:JANplosmed05.886", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Johns Hopkins Alzheimer's Disease Research Center" }, { "agency": "National Alliance for Research on Schizophrenia and Depression" }, { "agency": "Rose Hills Foundation" }, { "agency": "Alzheimer's Association" }, { "agency": "NIH", "grant_number": "K01 AG26144-01" }, { "agency": "NIH", "grant_number": "P50 AGO5146-20" }, { "agency": "NIH", "grant_number": "R01 AG006656-16" }, { "agency": "NIH", "grant_number": "P01 AG015453" }, { "agency": "NIH", "grant_number": "R01 NS 047225" }, { "agency": "National Cancer Institute" }, { "agency": "National Institute on Aging" } ] }, "doi": "10.1371/journal.pmed.0020355", "pmcid": "PMC1283364", "primary_object": { "basename": "JANplosmed05.pdf", "url": "https://authors.library.caltech.edu/records/5pmf7-4rg66/files/JANplosmed05.pdf" }, "related_objects": [ { "basename": "journal.pmed.0020355.sg001.TIF", "url": "https://authors.library.caltech.edu/records/5pmf7-4rg66/files/journal.pmed.0020355.sg001.TIF" }, { "basename": "journal.pmed.0020355.sg002.PSD", "url": "https://authors.library.caltech.edu/records/5pmf7-4rg66/files/journal.pmed.0020355.sg002.PSD" }, { "basename": "journal.pmed.0020355.sg003.PSD", "url": "https://authors.library.caltech.edu/records/5pmf7-4rg66/files/journal.pmed.0020355.sg003.PSD" }, { "basename": "journal.pmed.0020355.sg004.JPG", "url": "https://authors.library.caltech.edu/records/5pmf7-4rg66/files/journal.pmed.0020355.sg004.JPG" }, { "basename": "journal.pmed.0020355.sg005.TIF", "url": "https://authors.library.caltech.edu/records/5pmf7-4rg66/files/journal.pmed.0020355.sg005.TIF" }, { "basename": "journal.pmed.0020355.sg006.PSD", "url": "https://authors.library.caltech.edu/records/5pmf7-4rg66/files/journal.pmed.0020355.sg006.PSD" } ], "resource_type": "article", "pub_year": "2005", "author_list": "Jankowsky, Joanna L.; Slunt, Hilda H.; et el." }, { "id": "https://authors.library.caltech.edu/records/72051-jt503", "eprint_id": 55917, "eprint_status": "archive", "datestamp": "2023-08-19 16:38:58", "lastmod": "2023-10-20 23:25:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "Sarah C. R." }, "orcid": "0000-0001-9410-9805" }, { "id": "Beene-D-L", "name": { "family": "Beene", "given": "Darren L." } }, { "id": "Lee-Lori-W", "name": { "family": "Lee", "given": "Lori W." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Broadhurst-R-W", "name": { "family": "Broadhurst", "given": "R. William" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Cis\u2013trans isomerization at a proline opens the pore of a neurotransmitter-gated ion channel", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2005 Nature Publishing Group.\n\nReceived 24 March; accepted 8 August 2005.\n\nS.C.R.L. is a Wellcome Trust Senior Research Fellow in Basic Biomedical Science. The work at Caltech was supported by the National Institutes of Health. We thank K. L. Price and A. J. Thompson for assistance with modelling.\n\nSupplemental Material - nature04130-s1.pdf
", "abstract": "5-Hydroxytryptamine type 3 (5-HT_3) receptors are members of the Cys-loop receptor superfamily. Neurotransmitter binding in these proteins triggers the opening (gating) of an ion channel by means of an as-yet-uncharacterized conformational change. Here we show that a specific proline (Pro 8*), located at the apex of the loop between the second and third transmembrane helices (M2\u2013M3), can link binding to gating through a cis\u2013trans isomerization of the protein backbone. Using unnatural amino acid mutagenesis, a series of proline analogues with varying preference for the cis conformer was incorporated at the 8* position. Proline analogues that strongly favour the trans conformer produced non-functional channels. Among the functional mutants there was a strong correlation between the intrinsic cis\u2013trans energy gap of the proline analogue and the activation of the channel, suggesting that cis\u2013trans isomerization of this single proline provides the switch that interconverts the open and closed states of the channel. Consistent with this proposal, nuclear magnetic resonance studies on an M2\u2013M3 loop peptide reveal two distinct, structured forms. Our results thus confirm the structure of the M2\u2013M3 loop and the critical role of Pro 8* in the 5-HT_3 receptor. In addition, they suggest that a molecular rearrangement at Pro 8* is the structural mechanism that opens the receptor pore.", "date": "2005-11-10", "date_type": "published", "publication": "Nature", "volume": "438", "number": "4065", "publisher": "Nature Publishing Group", "pagerange": "248-252", "id_number": "CaltechAUTHORS:20150319-085128410", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150319-085128410", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "NIH" } ] }, "doi": "10.1038/nature04130", "primary_object": { "basename": "nature04130-s1.pdf", "url": "https://authors.library.caltech.edu/records/72051-jt503/files/nature04130-s1.pdf" }, "resource_type": "article", "pub_year": "2005", "author_list": "Lummis, Sarah C. R.; Beene, Darren L.; et el." }, { "id": "https://authors.library.caltech.edu/records/azvry-3vc57", "eprint_id": 102766, "eprint_status": "archive", "datestamp": "2023-08-19 16:20:57", "lastmod": "2023-10-20 00:29:15", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "Sarah C. R." }, "orcid": "0000-0001-9410-9805" }, { "id": "Beene-D-L", "name": { "family": "Beene", "given": "Darren L." } }, { "id": "Harrison-N-J", "name": { "family": "Harrison", "given": "Neil J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "A Cation-\u03c0 Binding Interaction with a Tyrosine in the Binding Site of the GABA_C Receptor", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2005 Elsevier Ltd. \n\nReceived 22 April 2005, Revised 27 June 2005, Accepted 28 June 2005, Available online 23 September 2005. \n\nWe would like to thank The Wellcome Trust (S.C.R.L. is a Wellcome Trust Senior Research Fellow in Basic Biomedical Science), the Medical Research Council (a studentship to N.J.H.), and the U.S. National Institutes of Health (NS11756, NS34407).", "abstract": "GABA_C (\u03c1) receptors are members of the Cys-loop superfamily of neurotransmitter receptors, which includes nicotinic acetylcholine (nACh), 5-HT\u2083, and glycine receptors. As in other members of this family, the agonist binding site of GABA_C receptors is rich in aromatic amino acids, but while other receptors bind agonist through a cation-\u03c0 interaction to a tryptophan, the GABA_C binding site has tyrosine at the aligning positions. Incorporating a series of tyrosine derivatives at position 198 using unnatural amino acid mutagenesis reveals a clear correlation between the cation-\u03c0 binding ability of the side chain and EC\u2085\u2080 for receptor activation, thus demonstrating a cation-\u03c0 interaction between a tyrosine side chain and a neurotransmitter. Comparisons among four homologous receptors show variations in cation-\u03c0 binding energies that reflect the nature of the cationic center of the agonist.", "date": "2005-09", "date_type": "published", "publication": "Chemistry and Biology", "volume": "12", "number": "9", "publisher": "Elsevier", "pagerange": "993-997", "id_number": "CaltechAUTHORS:20200424-080629607", "issn": "1074-5521", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200424-080629607", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "Medical Research Council (UK)" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" } ] }, "doi": "10.1016/j.chembiol.2005.06.012", "resource_type": "article", "pub_year": "2005", "author_list": "Lummis, Sarah C. R.; Beene, Darren L.; et el." }, { "id": "https://authors.library.caltech.edu/records/ejm89-gwg23", "eprint_id": 102875, "eprint_status": "archive", "datestamp": "2023-08-19 16:11:27", "lastmod": "2023-10-20 00:35:17", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Rodrigues-Pinguet-Nivalda-O", "name": { "family": "Rodrigues-Pinguet", "given": "Nivalda O." } }, { "id": "Pinguet-Thierry-J", "name": { "family": "Pinguet", "given": "Thierry J." }, "orcid": "0000-0002-9994-6482" }, { "id": "Figl-Antonio", "name": { "family": "Figl", "given": "Antonio" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce N." } } ] }, "title": "Mutations Linked to Autosomal Dominant Nocturnal Frontal Lobe Epilepsy Affect Allosteric Ca\u00b2\u207a Activation of the \u03b14\u03b22 Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2005 The American Society for Pharmacology and Experimental Therapeutics. \n\nReceived January 18, 2005; accepted May 17, 2005. \n\nThis work was supported by grants from the National Institute of Health (NS0438000 and NS011756, to B.N.C. and H.A.L., respectively and a minority supplement (NS0438000-04S1 to N.O.R.). \n\nPortions of this work were published previously by UMI publishers in a doctoral dissertation by N.O.R.\n\nSupplemental Material - Supplementary_Data.doc
", "abstract": "Extracellular Ca\u00b2\u207a robustly potentiates the acetylcholine response of \u03b14\u03b22 nicotinic receptors. Rat orthologs of five mutations linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE)\u2014\u03b14(S252F), \u03b14(S256L), \u03b14(+L264), \u03b22(V262L), and \u03b22(V262M)\u2014reduced 2 mM Ca\u00b2\u207a potentiation of the \u03b14\u03b22 1 mM acetylcholine response by 55 to 74%. To determine whether altered allosteric Ca\u00b2\u207a activation or enhanced Ca\u00b2\u207a block caused this reduction, we coexpressed the rat ADNFLE mutations with an \u03b14 N-terminal mutation, \u03b14(E180Q), that abolished \u03b14\u03b22 allosteric Ca\u00b2\u207a activation. In each case, Ca\u00b2\u207a inhibition of the double mutants was less than that expected from a Ca\u00b2\u207a blocking mechanism. In fact, the effects of Ca\u00b2\u207a on the ADNFLE mutations near the intracellular end of the M2 region\u2014\u03b14(S252F) and \u03b14(S256L)\u2014were consistent with a straightforward allosteric mechanism. In contrast, the effects of Ca\u00b2\u207a on the ADNFLE mutations near the extracellular end of the M2 region\u2014\u03b14(+L264)\u03b22, \u03b22(V262L), and \u03b22(V262M)\u2014were consistent with a mixed mechanism involving both altered allosteric activation and enhanced block. However, the effects of 2 mM Ca\u00b2\u207a on the \u03b14\u03b22, \u03b14(+L264)\u03b22, and \u03b14\u03b22(V262L) single-channel conductances, the effects of membrane potential on the \u03b22(V262L)-mediated reduction in Ca\u00b2\u207a potentiation, and the effects of eliminating the negative charges in the extracellular ring on this reduction failed to provide any direct evidence of mutant-enhanced Ca\u00b2\u207a block. Moreover, analyses of the \u03b14\u03b22, \u03b14(S256L), and \u03b14(+L264) Ca\u00b2\u207a concentration-potentiation relations suggested that the ADNFLE mutations reduce Ca\u00b2\u207a potentiation of the \u03b14\u03b22 acetylcholine response by altering allosteric activation rather than by enhancing block.", "date": "2005-08", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "68", "number": "2", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "487-501", "id_number": "CaltechAUTHORS:20200428-121436325", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200428-121436325", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS0438000" }, { "agency": "NIH", "grant_number": "NS011756" }, { "agency": "NIH", "grant_number": "NS0438000-04S1" } ] }, "doi": "10.1124/mol.105.011155", "primary_object": { "basename": "Supplementary_Data.doc", "url": "https://authors.library.caltech.edu/records/ejm89-gwg23/files/Supplementary_Data.doc" }, "resource_type": "article", "pub_year": "2005", "author_list": "Rodrigues-Pinguet, Nivalda O.; Pinguet, Thierry J.; et el." }, { "id": "https://authors.library.caltech.edu/records/v25xe-pqr61", "eprint_id": 102638, "eprint_status": "archive", "datestamp": "2023-08-22 03:57:20", "lastmod": "2023-10-20 00:22:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Khakh-B-S", "name": { "family": "Khakh", "given": "Baljit S." } }, { "id": "Fisher-J-A", "name": { "family": "Fisher", "given": "James A." } }, { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Bowser-D-N", "name": { "family": "Bowser", "given": "David N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "An Angstrom Scale Interaction between Plasma Membrane ATP-Gated P2X\u2082 and \u03b1\u2084\u03b2\u2082 Nicotinic Channels Measured with Fluorescence Resonance Energy Transfer and Total Internal Reflection Fluorescence Microscopy", "ispublished": "pub", "full_text_status": "public", "keywords": "channel; cholinergic; purinergic; acetylcholine receptor; ACh; fluorescence microscopy; P2X", "note": "\u00a9 2005 Society for Neuroscience. \n\nReceived Feb. 10, 2005; revised June 13, 2005; accepted June 14, 2005. \n\nResearch in our laboratories is supported by the Medical Research Council (MRC), the European Molecular Biology Organization, the Human Frontiers Science Program, and National Institutes of Health Grants NS11756 and DA17279. J.A.F. was supported by an MRC Studentship, R.N. was supported by a postdoctoral fellowship from the California Tobacco-Related Disease Research Program and the Elizabeth Ross Fund, and D.N.B. was supported in part by a European Molecular Biology Organization Fellowship. We thank Julian Revie for constructing some of the nicotinic ACh receptor cDNAs.\n\nPublished - 6911.full.pdf
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", "abstract": "Structurally distinct nicotinic and P2X channels interact functionally, such that coactivation results in cross-inhibition of one or both channel types. It is hypothesized, but not yet proven, that nicotinic and P2X channels interact at the plasma membrane. Here, we show that plasma membrane \u03b1\u2084\u03b2\u2082 nicotinic and P2X\u2082 channels form a molecular scale partnership and also influence each other when coactivated, resulting in nonadditive cross-inhibitory responses. Total internal reflection fluorescence and fluorescence resonance energy transfer microscopy between fluorescently labeled P2X\u2082 and \u03b1\u2084\u03b2\u2082 nicotinic channels demonstrated close spatial arrangement of the channels in human embryonic kidney cells and in hippocampal neuron membranes. The data suggest that P2X\u2082 and \u03b1\u2084\u03b2\u2082 channels may form a dimer, with the channels \u223c80 \u00c5 apart. The measurements also show that P2X\u2082 subunits interact specifically and robustly with the \u03b2\u2082 subunits in \u03b1\u2084\u03b2\u2082 channels. The data provide direct evidence for the close spatial apposition of full-length P2X\u2082 and \u03b1\u2084\u03b2\u2082 channels within 100 nm of the plasma membrane of living cells.", "date": "2005-07-20", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "25", "number": "29", "publisher": "Society for Neuroscience", "pagerange": "6911-6920", "id_number": "CaltechAUTHORS:20200417-142718484", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200417-142718484", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Medical Research Council (UK)" }, { "agency": "European Molecular Biology Organization (EMBO)" }, { "agency": "Human Frontier Science Program" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "DA17279" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Elizabeth Ross Fellowship" } ] }, "doi": "10.1523/jneurosci.0561-05.2005", "pmcid": "PMC6725363", "primary_object": { "basename": "6911.full.pdf", "url": "https://authors.library.caltech.edu/records/v25xe-pqr61/files/6911.full.pdf" }, "related_objects": [ { "basename": "fig1.gif", "url": "https://authors.library.caltech.edu/records/v25xe-pqr61/files/fig1.gif" }, { "basename": "fig2.gif", "url": "https://authors.library.caltech.edu/records/v25xe-pqr61/files/fig2.gif" } ], "resource_type": "article", "pub_year": "2005", "author_list": "Khakh, Baljit S.; Fisher, James A.; et el." }, { "id": "https://authors.library.caltech.edu/records/9b4f1-79y33", "eprint_id": 103014, "eprint_status": "archive", "datestamp": "2023-08-22 03:23:02", "lastmod": "2023-10-20 00:42:08", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Chiu-Chi-Sung", "name": { "family": "Chiu", "given": "Chi-Sung" } }, { "id": "Brickley-S", "name": { "family": "Brickley", "given": "Stephen" } }, { "id": "Jensen-Kimmo", "name": { "family": "Jensen", "given": "Kimmo" } }, { "id": "Southwell-A-L", "name": { "family": "Southwell", "given": "Amber" } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Cull-Candy-S", "name": { "family": "Cull-Candy", "given": "Stuart" } }, { "id": "Mody-I", "name": { "family": "Mody", "given": "Istvan" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "GABA Transporter Deficiency Causes Tremor, Ataxia, Nervousness, and Increased GABA-Induced Tonic Conductance in Cerebellum", "ispublished": "pub", "full_text_status": "public", "keywords": "tiagabine; epilepsy; flunitrazepam; cerebellum; inhibition; tremor", "note": "\u00a9 2005 Society for Neuroscience. \n\nReceived Aug. 16, 2004; revised Jan. 25, 2005; accepted Jan. 25, 2005. \n\nThis research was supported by National Institutes of Health Grants DA-01921, NS-11756, MH-49176, NS-030549, and DA-010509, National Science Foundation Grant 0119493, the Wellcome Trust, a Royal Society-Wolfson Award (S.C.-C.), and a Della Martin Fellowship (C.-S.C.). We are indebted to members of Caltech and University of California Los Angeles groups for advice, Limin Shi and Paul Patterson for use and help with the startle system, and J. Crawley for comments on this manuscript.\n\nPublished - 3234.full.pdf
Supplemental Material - Chiu-04-supplementary-figur.gif
", "abstract": "GABA transporter subtype 1 (GAT1) knock-out (KO) mice display normal reproduction and life span but have reduced body weight (female, -10%; male, -20%) and higher body temperature fluctuations in the 0.2-1.5/h frequency range. Mouse GAT1 (mGAT1) KO mice exhibit motor disorders, including gait abnormality, constant 25-32 Hz tremor, which is aggravated by flunitrazepam, reduced rotarod performance, and reduced locomotor activity in the home cage. Open-field tests show delayed exploratory activity, reduced rearing, and reduced visits to the central area, with no change in the total distance traveled. The mGAT1 KO mice display no difference in acoustic startle response but exhibit a deficiency in prepulse inhibition. These open-field and prepulse inhibition results suggest that the mGAT1 KO mice display mild anxiety or nervousness. The compromised GABA uptake in mGAT1 KO mice results in an increased GABA_A receptor-mediated tonic conductance in both cerebellar granule and Purkinje cells. The reduced rate of GABA clearance from the synaptic cleft is probably responsible for the slower decay of spontaneous IPSCs in cerebellar granule cells. There is little or no compensatory change in other proteins or structures related to GABA transmission in the mGAT1 KO mice, including GAT1-independent GABA uptake, number of GABAergic interneurons, and GABA_A-, vesicular GABA transporter-, GAD65-, and GAT3-immunoreactive structures in cerebellum or hippocampus. Therefore, the excessive extracellular GABA present in mGAT1 KO mice results in behaviors that partially phenocopy the clinical side effects of tiagabine, suggesting that these side effects are inherent to a therapeutic strategy that targets the widely expressed GAT1 transporter system.", "date": "2005-03-23", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "25", "number": "12", "publisher": "Society for Neuroscience", "pagerange": "3234-3245", "id_number": "CaltechAUTHORS:20200506-072848149", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200506-072848149", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-01921" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "NS-030549" }, { "agency": "NIH", "grant_number": "DA-010509" }, { "agency": "NSF", "grant_number": "CBET-0119493" }, { "agency": "Wellcome Trust" }, { "agency": "Royal Society" }, { "agency": "Della Martin Foundation" } ] }, "doi": "10.1523/jneurosci.3364-04.2005", "pmcid": "PMC6725086", "primary_object": { "basename": "3234.full.pdf", "url": "https://authors.library.caltech.edu/records/9b4f1-79y33/files/3234.full.pdf" }, "related_objects": [ { "basename": "Chiu-04-supplementary-figur.gif", "url": "https://authors.library.caltech.edu/records/9b4f1-79y33/files/Chiu-04-supplementary-figur.gif" } ], "resource_type": "article", "pub_year": "2005", "author_list": "Chiu, Chi-Sung; Brickley, Stephen; et el." }, { "id": "https://authors.library.caltech.edu/records/sh4dz-exd75", "eprint_id": 103015, "eprint_status": "archive", "datestamp": "2023-08-22 03:13:38", "lastmod": "2023-10-20 00:42:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kovoor-A", "name": { "family": "Kovoor", "given": "Abraham" } }, { "id": "Seyffarth-P", "name": { "family": "Seyffarth", "given": "Petra" } }, { "id": "Ebert-J", "name": { "family": "Ebert", "given": "Jana" } }, { "id": "Barghshoon-S", "name": { "family": "Barghshoon", "given": "Sami" } }, { "id": "Chen-Ching-Kang", "name": { "family": "Chen", "given": "Ching-Kang" } }, { "id": "Schwarz-S", "name": { "family": "Schwarz", "given": "Sigrid" } }, { "id": "Axelrod-J-D", "name": { "family": "Axelrod", "given": "Jeffrey D." } }, { "id": "Cheyette-B-N-R", "name": { "family": "Cheyette", "given": "Benjamin N. R." } }, { "id": "Simon-M-I", "name": { "family": "Simon", "given": "Melvin I." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Schwarz-J", "name": { "family": "Schwarz", "given": "Johannes" } } ] }, "title": "D\u2082 Dopamine Receptors Colocalize Regulator of G-Protein Signaling 9-2 (RGS9-2) via the RGS9 DEP Domain, and RGS9 Knock-Out Mice Develop Dyskinesias Associated with Dopamine Pathways", "ispublished": "pub", "full_text_status": "public", "keywords": "antipsychotic; D2 dopamine receptor; dyskinesia; DEP domain; RGS9; striatum", "note": "\u00a9 2005 Society for Neuroscience. \n\nReceived Jan. 15, 2004; revised Jan. 4, 2005; accepted Jan. 4, 2005. \n\nThis work was supported by grants from the Interdisziplin\u00e4res Zentrum f\u00fcr klinische Forschung Leipzig (TP C19), National Institutes of Health (GM-29836, GM05982, MH001750, and MH-49176), and by a National Research Service Award to A.K. (MH019552).\n\nPublished - 2157.full.pdf
", "abstract": "Regulator of G-protein signaling 9-2 (RGS9-2), a member of the RGS family of G\u03b1 GTPase accelerating proteins, is expressed specifically in the striatum, which participates in antipsychotic-induced tardive dyskinesia and in levodopa-induced dyskinesia. We report that RGS9 knock-out mice develop abnormal involuntary movements when inhibition of dopaminergic transmission is followed by activation of D\u2082-like dopamine receptors (DRs). These abnormal movements resemble drug-induced dyskinesia more closely than other rodent models. Recordings from striatal neurons of these mice establish that activation of D\u2082-like DRs abnormally inhibits glutamate-elicited currents. We show that RGS9-2, via its DEP domain (for Disheveled, EGL-10, Pleckstrin homology), colocalizes with D\u2082DRs when coexpressed in mammalian cells. Recordings from oocytes coexpressing D\u2082DR or the m2 muscarinic receptor and G-protein-gated inward rectifier potassium channels show that RGS9-2, via its DEP domain, preferentially accelerates the termination of D\u2082DR signals. Thus, alterations in RGS9-2 may be a key factor in the pathway leading from D\u2082DRs to the side effects associated with the treatment both of psychoses and Parkinson's disease.", "date": "2005-02-23", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "25", "number": "8", "publisher": "Society for Neuroscience", "pagerange": "2157-2165", "id_number": "CaltechAUTHORS:20200506-074200832", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200506-074200832", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Interdisziplin\u00e4res Zentrum f\u00fcr klinische Forschung Leipzig", "grant_number": "TP C19" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "GM05982" }, { "agency": "NIH", "grant_number": "MH001750" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "MH019552" } ] }, "doi": "10.1523/jneurosci.2840-04.2005", "pmcid": "PMC6726050", "primary_object": { "basename": "2157.full.pdf", "url": "https://authors.library.caltech.edu/records/sh4dz-exd75/files/2157.full.pdf" }, "resource_type": "article", "pub_year": "2005", "author_list": "Kovoor, Abraham; Seyffarth, Petra; et el." }, { "id": "https://authors.library.caltech.edu/records/8r046-5my74", "eprint_id": 76902, "eprint_status": "archive", "datestamp": "2023-08-19 15:15:31", "lastmod": "2023-10-25 16:58:20", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Cashin-A-L", "name": { "family": "Cashin", "given": "Amanda L." } }, { "id": "Petersson-E-J", "name": { "family": "Petersson", "given": "E. James" }, "orcid": "0000-0003-3854-9210" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Using Physical Chemistry To Differentiate Nicotinic from Cholinergic Agonists at the Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2005 American Chemical Society. \n\nReceived June 28, 2004. Publication Date (Web): December 2, 2004. \n\nWe thank the NIH (NS 34407 and NS 11756) for support of this work, and Professor George Petersson for assistance.\n\nSupplemental Material - ja0461771si20040902_042520.pdf
", "abstract": "The binding of three distinct agonists - acetylcholine (ACh), nicotine, and epibatidine - to the nicotinic acetylcholine receptor has been probed using unnatural amino acid mutagenesis. ACh makes a cation\u2212\u03c0 interaction with Trp \u03b1149, while nicotine employs a hydrogen bond to a backbone carbonyl in the same region of the agonist binding site. The nicotine analogue epibatidine achieves its high potency by taking advantage of both the cation\u2212\u03c0 interaction and the backbone hydrogen bond. A simple structural model that considers only possible interactions with Trp \u03b1149 suggests that a novel aromatic C - H\u00b7\u00b7\u00b7O=C hydrogen bond further augments the binding of epibatidine. These studies illustrate the subtleties and complexities of the interactions between drugs and membrane receptors and establish a paradigm for obtaining detailed structural information.", "date": "2005-01-12", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "127", "number": "1", "publisher": "American Chemical Society", "pagerange": "350-356", "id_number": "CaltechAUTHORS:20170425-083958251", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170425-083958251", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1021/ja0461771", "primary_object": { "basename": "ja0461771si20040902_042520.pdf", "url": "https://authors.library.caltech.edu/records/8r046-5my74/files/ja0461771si20040902_042520.pdf" }, "resource_type": "article", "pub_year": "2005", "author_list": "Cashin, Amanda L.; Petersson, E. James; et el." }, { "id": "https://authors.library.caltech.edu/records/crmbp-b7t91", "eprint_id": 51942, "eprint_status": "archive", "datestamp": "2023-08-19 14:39:50", "lastmod": "2023-10-18 18:43:55", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Tapper-A-R", "name": { "family": "Tapper", "given": "Andrew R." } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri L." } }, { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Schwarz-J", "name": { "family": "Schwarz", "given": "Johannes" } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Whiteaker-P", "name": { "family": "Whiteaker", "given": "Paul" } }, { "id": "Marks-M-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Collins-A-C", "name": { "family": "Collins", "given": "Allan C." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Nicotine Activation of \u03b14* Receptors: Sufficient for Reward, Tolerance, and Sensitization", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2004 American Association for the Advancement of Science.\n\n21 April 2004; accepted 2 September 2004.\n\nThis research was supported by the California Tobacco-Related Disease Research Project (grant 12RT-0245 and fellowship 10FT-0174 to R.N.), by the NIH (grants DA-3194 and DA-15663 at Boulder, NS-11756 and MH-49716 at Caltech, and National Research Service Award to A.R.T.), and by the W. M. Keck and Plum Foundations. We thank C. Fonck, B. Cohen, S. Kwoh, N. Rodrigues-Pinguet, S. Grady, J. Wehner, and S. Malin for access to unpublished work.\n\nSupplemental Material - Tapper.SOM.pdf
", "abstract": "The identity of nicotinic receptor subtypes sufficient to elicit both the acute and chronic effects of nicotine dependence is unknown. We engineered mutant mice with \u03b14 nicotinic subunits containing a single point mutation, Leu^(9\u2032) \u2192 Ala^(9\u2032) in the pore-forming M2 domain, rendering \u03b14* receptors hypersensitive to nicotine. Selective activation of \u03b14* nicotinic acetylcholine receptors with low doses of agonist recapitulates nicotine effects thought to be important in dependence, including reinforcement in response to acute nicotine administration, as well as tolerance and sensitization elicited by chronic nicotine administration. These data indicate that activation of \u03b14* receptors is sufficient for nicotine-induced reward, tolerance, and sensitization.", "date": "2004-11-05", "date_type": "published", "publication": "Science", "volume": "306", "number": "5698", "publisher": "American Association for the Advancement of Science", "pagerange": "1029-1032", "id_number": "CaltechAUTHORS:20141119-090016448", "issn": "0036-8075", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20141119-090016448", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "12RT-0245" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "10FT-0174" }, { "agency": "NIH", "grant_number": "DA-3194" }, { "agency": "NIH", "grant_number": "DA-15663" }, { "agency": "NIH", "grant_number": "NS- 11756" }, { "agency": "NIH", "grant_number": "MH-49716" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "W. M. Keck Foundation" }, { "agency": "Plum Foundation" } ] }, "doi": "10.1126/science.1099420", "primary_object": { "basename": "Tapper.SOM.pdf", "url": "https://authors.library.caltech.edu/records/crmbp-b7t91/files/Tapper.SOM.pdf" }, "resource_type": "article", "pub_year": "2004", "author_list": "Tapper, Andrew R.; McKinney, Sheri L.; et el." }, { "id": "https://authors.library.caltech.edu/records/wwrw2-6bv60", "eprint_id": 40359, "eprint_status": "archive", "datestamp": "2023-09-15 04:06:02", "lastmod": "2023-10-23 21:01:05", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Diba-Kamran", "name": { "family": "Diba", "given": "Kamran" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Koch-C", "name": { "family": "Koch", "given": "Christof" }, "orcid": "0000-0001-6482-8067" } ] }, "title": "Intrinsic Noise in Cultured Hippocampal Neurons: Experiment and Modeling", "ispublished": "pub", "full_text_status": "public", "keywords": "voltage noise; current noise; biophysical modeling; coding; neural computation; PSD", "note": "\u00a92004 Society for Neuroscience. \n\nReceived March 15, 2004. Revision received August 12, 2004. Accepted August 13, 2004. \n\nThis work was supported by the National Institute of Mental Health, the National Science Foundation, and a Sloan-Swartz Postdoctoral Fellowship to K.D. We are deeply indebted to Irina Sokolova and Sheri McKinney for help with the culture preparation and patch clamp. Additionally,wethank Eric Slimko, Sacha Malin, Gilad Jacobson, Yosef Yarom, and Idan Segev for valuable discussions and Michael Hines and Ted Carnevale for assistance with the NEURON simulation environment.\n\nPublished - 502.pdf
", "abstract": "Ion channels open and close stochastically. The fluctuation of these channels represents an intrinsic source of noise that affects the input-output properties of the neuron. We combined whole-cell measurements with biophysical modeling to characterize the intrinsic stochastic and electrical properties of single neurons as observed at the soma. We measured current and voltage noise in 18 d postembryonic cultured neurons from the rat hippocampus, at various subthreshold and near-threshold holding potentials in the presence of synaptic blockers. The observed current noise increased with depolarization, as ion channels were activated, and its spectrum demonstrated generalized 1/fbehavior. Exposure to TTX removed a significant contribution from Na^+ channels to the noise spectrum, particularly at depolarized potentials, and the resulting spectrum was now dominated by a single Lorentzian (1/f^2) component. By replacing the intracellular K^+ with Cs^+, we demonstrated that a major portion of the observed noise was attributable to K^+ channels. We compared the measured power spectral densities to a 1-D cable model of channel fluctuations based on Markov kinetics. We found that a somatic compartment, in combination with a single equivalent cylinder, described the effective geometry from the viewpoint of the soma. Four distinct channel populations were distributed in the membrane and modeled as Lorentzian current noise sources. Using the NEURON simulation program, we summed up the contributions from the spatially distributed current noise sources and calculated the total voltage and current noise. Our quantitative model reproduces important voltage- and frequency-dependent features of the data, accounting for the 1/f behavior, as well as the effects of various blockers.", "date": "2004-10-27", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "24", "number": "43", "publisher": "Society for Neuroscience", "pagerange": "9723-9733", "id_number": "CaltechAUTHORS:20130816-103138809", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20130816-103138809", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Institute of Mental Health (NIMH)" }, { "agency": "NSF" }, { "agency": "Sloan-Swartz Foundation" }, { "agency": "NIH" } ] }, "local_group": { "items": [ { "id": "Koch-Laboratory" } ] }, "doi": "10.1523/JNEUROSCI.1721-04.2004", "pmcid": "PMC6730144", "primary_object": { "basename": "502.pdf", "url": "https://authors.library.caltech.edu/records/wwrw2-6bv60/files/502.pdf" }, "resource_type": "article", "pub_year": "2004", "author_list": "Diba, Kamran; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/s1e68-hta26", "eprint_id": 102788, "eprint_status": "archive", "datestamp": "2023-08-22 02:33:26", "lastmod": "2023-10-20 00:30:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Beene-D-L", "name": { "family": "Beene", "given": "Darren L." } }, { "id": "Price-K-L", "name": { "family": "Price", "given": "Kerry L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lummis-S-C-R", "name": { "family": "Lummis", "given": "Sarah C. R." }, "orcid": "0000-0001-9410-9805" } ] }, "title": "Tyrosine Residues That Control Binding and Gating in the 5-Hydroxytryptamine\u2083 Receptor Revealed by Unnatural Amino Acid Mutagenesis", "ispublished": "pub", "full_text_status": "public", "keywords": "ligand-gated ion channel; Cys-loop receptor; 5-HT3 receptor binding site; hydrogen bond; unnatural amino acids; activation\nmechanism; serotonin", "note": "\u00a9 2004 Society for Neuroscience. \n\nReceived April 23, 2004; accepted July 20, 2004. \n\nThis work was supported by The Wellcome Trust (S.C.R.L. is a Wellcome Trust Senior Research Fellow in Basic Biomedical Science), the Medical Research Council (a studentship to K.L.P.), and the National Institutes of Health (Grants NS11756 and NS34407). We thank Dr. Nigel Unwin for helpful discussions.\n\nPublished - 9097.full.pdf
", "abstract": "The mechanism by which agonist binding triggers pore opening in ligand-gated ion channels is poorly understood. Here, we used unnatural amino acid mutagenesis to introduce subtle changes to the side chains of tyrosine residues (Tyr141, Tyr143, Tyr153, and Tyr234), which dominate the 5-HT\u2083 receptor binding site. Heterologous expression in oocytes, combined with radioligand binding data and a model of 5-HT (serotonin) computationally docked into the binding site, has allowed us to determine which of these residues are responsible for binding and/or gating. We have shown that Tyr 143 forms a hydrogen bond that is essential for receptor gating but does not affect binding, whereas a hydrogen bond formed by Tyr153 is involved in both binding and gating of the receptor. The aromatic group of Tyr234 is essential for binding and gating, whereas its hydroxyl does not affect binding but plays a steric role in receptor gating. Tyr141 is not involved in agonist binding or receptor gating but is important for antagonist interactions. These data, combined with a new model of the nonliganded 5-HT\u2083 receptor, lead to a mechanistic explanation of the interactions that initiate the conformational change leading to channel opening. Thus, we suggest that agonist entry into the binding pocket may displace Tyr143 and Tyr153 and results in their forming new hydrogen bonds. These bonds may form part of the network of bond rearrangements that trigger the conformational change leading to channel opening. Similar rearrangements may initiate gating in all Cys-loop receptors.", "date": "2004-10-13", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "24", "number": "41", "publisher": "Society for Neuroscience", "pagerange": "9097-9104", "id_number": "CaltechAUTHORS:20200427-084524229", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-084524229", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "Medical Research Council (UK)" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" } ] }, "doi": "10.1523/jneurosci.2429-04.2004", "pmcid": "PMC6730062", "primary_object": { "basename": "9097.full.pdf", "url": "https://authors.library.caltech.edu/records/s1e68-hta26/files/9097.full.pdf" }, "resource_type": "article", "pub_year": "2004", "author_list": "Beene, Darren L.; Price, Kerry L.; et el." }, { "id": "https://authors.library.caltech.edu/records/qxv9k-0xq39", "eprint_id": 73843, "eprint_status": "archive", "datestamp": "2023-08-19 14:10:52", "lastmod": "2023-10-24 21:01:59", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Becker-C-F-W", "name": { "family": "Becker", "given": "Christian F. W." } }, { "id": "Clayton-D", "name": { "family": "Clayton", "given": "Daniel" } }, { "id": "Shapovalov-G", "name": { "family": "Shapovalov", "given": "George" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Kochendoerfer-G-G", "name": { "family": "Kochendoerfer", "given": "Gerd G." } } ] }, "title": "On-Resin Assembly of a Linkerless Lanthanide(III)-Based Luminescence Label and Its Application to the Total Synthesis of Site-Specifically Labeled Mechanosensitive Channels", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2004 American Chemical Society. \n\nReceived 19 May 2004. Revised Manuscript Received July 15, 2004. Published online 24 August 2004. Published in print 1 September 2004. \n\nThis work was supported by an NIH Program Project Grant, GM-062532.", "abstract": "A synthesis strategy for the on-resin assembly of luminescent lanthanide chelates from commercially available compounds was developed. Advantages of the approach include the absence of spacers between the metal ion and the attachment site, and the compatibility with typical chemical protein synthesis protection schemes. Methoxycoumarin-labeled lysine and tris(tert-butyl)-DOTA were consecutively coupled with high efficiency to a free amino group in otherwise fully protected peptide segments using standard peptide synthesis methods. Addition of stoichiometric amounts of Tb3+ to the modified, cleaved, and purified peptides yielded the desired lanthanide chelate. Incorporation of this label into a chemically synthesized, full-length mechanosensitive channel of large conductance (MscL) of E. coli and subsequent reconstitution into vesicles resulted in a functional mechanosensitive channel of comparable conductance to the wild-type channel. However, this channel required increased suction to gate. Excitation of the antenna molecule methoxycoumarin at 336 nm resulted in an emission spectrum typical for Tb3+ and a luminescence lifetime of 0.67 ms. The location of the probe close to the backbone of this protein may provide precise information about conformational changes during channel opening from LRET studies.", "date": "2004-09", "date_type": "published", "publication": "Bioconjugate Chemistry", "volume": "15", "number": "5", "publisher": "American Chemical Society", "pagerange": "1118-1124", "id_number": "CaltechAUTHORS:20170131-075938701", "issn": "1043-1802", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170131-075938701", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-062532" } ] }, "doi": "10.1021/bc0498828", "resource_type": "article", "pub_year": "2004", "author_list": "Becker, Christian F. W.; Clayton, Daniel; et el." }, { "id": "https://authors.library.caltech.edu/records/4px9k-s2086", "eprint_id": 102733, "eprint_status": "archive", "datestamp": "2023-08-22 02:17:51", "lastmod": "2023-10-20 00:27:06", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Orb-Sabine", "name": { "family": "Orb", "given": "Sabine" } }, { "id": "Wieacker-J", "name": { "family": "Wieacker", "given": "Johannes" } }, { "id": "Labarca-C-G", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Fonck-C", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Schwarz-J", "name": { "family": "Schwarz", "given": "Johannes" } } ] }, "title": "Knockin mice with Leu9\u2032Ser \u03b14-nicotinic receptors: substantia nigra dopaminergic neurons are hypersensitive to agonist and lost postnatally", "ispublished": "pub", "full_text_status": "restricted", "keywords": "cholinergic excitotoxicity; mouse", "note": "\u00a9 2004 the American Physiological Society. \n\nSubmitted 16 January 2004; accepted in final form 7 June 2004. \n\nThis work was supported by German Research Foundation Grant DFG 704/3-1, the Tobacco Related Disease Research Project of California, NS-11756 and MH-49176, and a National Research Service Award (to C. Fonck). \n\nWe thank Annett Brandt, Purnima Deshpande, and Ute Roemuss for technical assistance.", "abstract": "This study analyzes the electrophysiological cause and behavioral consequence of dopaminergic cell loss in a knockin mouse strain bearing hypersensitive nicotinic \u03b14-receptor subunits (\"L9\u2032S mice\"). Adult brains of L9\u2032S mice show moderate loss of substantia nigra dopaminergic neurons and of striatal dopaminergic innervation. Amphetamine-stimulated locomotion is impaired, reflecting a reduction of dopamine stored in presynaptic vesicles. Recordings from dopaminergic neurons in L9\u2032S mice show that 10 \u03bcM nicotine depolarizes cells and increases spiking rates in L9\u2032S cells but hyperpolarizes and decreases spiking rates in wild-type (WT) cells. Thus dopaminergic neurons of L9\u2032S mice have an excitatory response to nicotine which is qualitatively different from that of WT neurons. The cause of dopaminergic cell death is therefore probably an increased sensitivity to acetylcholine or choline of \u03b14-containing nicotinic receptors. Hypersensitive excitatory stimulation during activation of \u03b14-containing receptors provides the first evidence for cholinergic excitotoxicity as a cause of dopaminergic neuron death. This novel concept may be relevant to the pathophysiology of Parkinson disease.", "date": "2004-08-11", "date_type": "published", "publication": "Physiological Genomics", "volume": "18", "number": "3", "publisher": "American Physiological Society", "pagerange": "299-307", "id_number": "CaltechAUTHORS:20200422-150959788", "issn": "1094-8341", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200422-150959788", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Deutsche Forschungsgemeinschaft (DFG)", "grant_number": "704/3-1" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1152/physiolgenomics.00012.2004", "resource_type": "article", "pub_year": "2004", "author_list": "Orb, Sabine; Wieacker, Johannes; et el." }, { "id": "https://authors.library.caltech.edu/records/xdems-qwf09", "eprint_id": 6313, "eprint_status": "archive", "datestamp": "2023-08-22 02:13:47", "lastmod": "2023-10-16 20:15:50", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Shapalov-G", "name": { "family": "Shapalov", "given": "George" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Gating Transitions in Bacterial Ion Channels Measured at 3 \u00b5s Resolution", "ispublished": "pub", "full_text_status": "public", "keywords": "MscL; MscS; patch clamping; single channel; substate", "note": "\u00a9 2004 The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nSubmitted: 3 May 2004. Accepted: 23 June 2004. Published online Jul 26 2004. doi:10.1085/jgp.200409087 \n\nWe thank Fred Sigworth, Dennis Dougherty, and Doug Rees for guidance, Josef Dudel for instruction on optimizing headstages and quartz pipettes, Randal Bass and Yan Poon for protein samples, Lori Lee for providing unpublished data, Alan Finkel, Richard Lobdill, and Eric Fung for help with Axopatch and Digidata modifications, and Daniel Clayton for help with the manuscript and discussions. \n\nThis work was supported by a grant from the National Institutes of Health (GM-062532).\n\nPublished - SHAjgp04.pdf
", "abstract": "Ion channels of high conductance (>200 pS) are widespread among prokaryotes and eukaryotes. Two examples, the Escherichia coli mechanosensitive ion channels Ec-MscS and Ec-MscL, pass currents of 125\u2013300 pA. To resolve temporal details of conductance transitions, a patch-clamp setup was optimized for low-noise recordings at a time resolution of 3 \u00b5s (10\u201320 times faster than usual). Analyses of the high-resolution recordings confirm that Ec-MscL visits many subconductance states and show that most of the intersubstate transitions occur more slowly than the effective resolution of 3 \u00b5s. There is a clear trend toward longer transition times for the larger transitions. In Ec-MscS recordings, the majority of the observed full conductance transitions are also composite. We detected a short-lived (~20 \u00b5s) Ec-MscS substate at 2/3 of full conductance; transitions between 2/3 and full conductance did not show fine structure and had a time course limited by the achieved resolution. Opening and closing transitions in MscS are symmetrical and are not preceded or followed by smaller, rapid currents (\"anticipations\" or \"regrets\"). Compared with other, lower-conductance channels, these measurements may detect unusually early states in the transitions from fully closed to fully open. Increased temporal resolution at the single-molecule level reveals that some elementary steps of structural transitions are composite and follow several alternative pathways, while others still escape resolution. High-bandwidth, low-noise single-channel measurements may provide details about state transitions in other high-conductance channels; and similar procedures may also be applied to channel- and nanopore-based single-molecule DNA measurements.", "date": "2004-08", "date_type": "published", "publication": "Journal of General Physiology", "volume": "124", "number": "2", "publisher": "Rockefeller University Press", "pagerange": "151-161", "id_number": "CaltechAUTHORS:SHAjgp04", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:SHAjgp04", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-062532" } ] }, "doi": "10.1085/jgp.200409087", "pmcid": "PMC2229625", "primary_object": { "basename": "SHAjgp04.pdf", "url": "https://authors.library.caltech.edu/records/xdems-qwf09/files/SHAjgp04.pdf" }, "resource_type": "article", "pub_year": "2004", "author_list": "Shapalov, George and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/j3atx-wpa16", "eprint_id": 602, "eprint_status": "archive", "datestamp": "2023-08-22 02:09:21", "lastmod": "2023-10-13 21:53:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dahan-D-S", "name": { "family": "Dahan", "given": "David S." } }, { "id": "Dibas-M-I", "name": { "family": "Dibas", "given": "Mohammed I." } }, { "id": "Petersson-E-J", "name": { "family": "Petersson", "given": "E. James" }, "orcid": "0000-0003-3854-9210" }, { "id": "Auyeung-Vincent-C", "name": { "family": "Auyeung", "given": "Vincent C." }, "orcid": "0000-0001-6273-1595" }, { "id": "Chanda-B", "name": { "family": "Chanda", "given": "Baron" } }, { "id": "Bezanilla-F", "name": { "family": "Bezanilla", "given": "Francisco" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "A fluorophore attached to nicotinic acetylcholine receptor beta M2 detects productive binding of agonist to the alpha delta site", "ispublished": "pub", "full_text_status": "public", "keywords": "LIGAND-BINDING, CHANNEL, SUBUNIT, DESENSITIZATION, FLUORESCENCE, ACTIVATION, MECHANISM, AFFINITY, REVEALS", "note": "\u00a9 2004 by The National Academy of Sciences of the USA. \n\nPublished online before print June 24 2004. \n\nEdited by Arthur Karlin, Columbia University College of Physicians and Surgeons, New York, NY and approved April 19, 2004 (received for review April 1, 2003). \n\nWe thank A. Karlin for providing the Beta-A19'C mutant and K. Kostenko and S. Kwoh for assistance with oocytes. This work was supported by National Institutes of Health Grants NS-11756, NS-34407, and GM-30376, and by a fellowship from the American Heart Association (to B.C.).\n\nPublished - DAHpnas04.pdf
", "abstract": "To study conformational transitions at the muscle nicotinic acetylcholine (ACh) receptor (nAChR), a rhodamine fluorophore was tethered to a Cys side chain introduced at the beta-19' position in the M2 region of the nAChR expressed in Xenopus oocytes. This procedure led to only minor changes in receptor function. During agonist application, fluorescence increased by (Delta-F/F) approximate to 10%, and the emission peak shifted to lower wavelengths, indicating a more hydrophobic environment for the fluorophore. The dose-response relations for Delta-F agreed well with those for epibatidine-induced currents, but were shifted approximate to 100-fold to the left of those for ACh-induced currents. Because (i) epibatidine binds more tightly to the alpha-gamma-binding site than to the alpha-delta site and (ii) ACh binds with reverse-site selectivity, these data suggest that Delta-F monitors an event linked to binding specifically at the alpha-delta-subunit interface. In experiments with flash-applied agonists, the earliest detectable Delta-F occurs within milliseconds, i.e., during activation. At low [ACh] (less than or equal to 10 muM), a phase of Delta-F occurs with the same time constant as desensitization, presumably monitoring an increased population of agonist-bound receptors. However, recovery from Delta-F is complete before the slowest phase of recovery from desensitization (time constant approximate to 250 s), showing that one or more desensitized states have fluorescence like that of the resting channel. That conformational transitions at the alpha-delta-binding site are not tightly coupled to channel activation suggests that sequential rather than fully concerted transitions occur during receptor gating. Thus, time-resolved fluorescence changes provide a powerful probe of nAChR conformational changes.", "date": "2004-07-06", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "101", "number": "27", "publisher": "National Academy of Sciences", "pagerange": "10195-10200", "id_number": "CaltechAUTHORS:DAHpnas04", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:DAHpnas04", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "GM-30376" }, { "agency": "American Heart Association" } ] }, "doi": "10.1073/pnas.0301885101", "pmcid": "PMC454187", "primary_object": { "basename": "DAHpnas04.pdf", "url": "https://authors.library.caltech.edu/records/j3atx-wpa16/files/DAHpnas04.pdf" }, "resource_type": "article", "pub_year": "2004", "author_list": "Dahan, David S.; Dibas, Mohammed I.; et el." }, { "id": "https://authors.library.caltech.edu/records/q5zer-n0t52", "eprint_id": 103163, "eprint_status": "archive", "datestamp": "2023-08-22 02:04:48", "lastmod": "2023-10-20 15:43:17", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Bhattacharya-A", "name": { "family": "Bhattacharya", "given": "Anindya" } }, { "id": "Dang-Hong", "name": { "family": "Dang", "given": "Hong" } }, { "id": "Zhu-Quan-Ming", "name": { "family": "Zhu", "given": "Quan-Ming" } }, { "id": "Schnegelsberg-B", "name": { "family": "Schnegelsberg", "given": "Birthe" } }, { "id": "Rozengurt-N", "name": { "family": "Rozengurt", "given": "Nora" } }, { "id": "Cain-G", "name": { "family": "Cain", "given": "Gary" } }, { "id": "Prantil-R", "name": { "family": "Prantil", "given": "Rachelle" } }, { "id": "Vorp-D-A", "name": { "family": "Vorp", "given": "David A." } }, { "id": "Guy-N", "name": { "family": "Guy", "given": "Nicholas" } }, { "id": "Julius-D", "name": { "family": "Julius", "given": "David" } }, { "id": "Ford-A-P-D-W", "name": { "family": "Ford", "given": "Anthony P. D. W." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Cockayne-D-A", "name": { "family": "Cockayne", "given": "Debra A." } } ] }, "title": "Uropathic Observations in Mice Expressing a Constitutively Active Point Mutation in the 5-HT_(3A) Receptor Subunit", "ispublished": "pub", "full_text_status": "public", "keywords": "5-HT3; mouse; knock-in mutation; bladder; hypertrophy; afferent innervation", "note": "\u00a9 2004 Society for Neuroscience. \n\nReceived Dec. 22, 2003; revised March 18, 2004; accepted April 22, 2004. \n\nThis work was supported by National Institutes of Health (NIH) National Research Service Award (H.D.). Funding from NIH Grant NS11756 is gratefully acknowledged. We thank E. Saclolo (Roche), C. Shilyansky, A. Tapper, A. Southwell, and C. Lindsell for help with maintaining the mice.\n\nPublished - 5537.full.pdf
", "abstract": "Mutant mice with a hypersensitive serotonin (5-HT)_(3A) receptor were generated through targeted exon replacement. A valine to serine mutation (V13\u2032S) in the channel-lining M2 domain of the 5-HT_(3A) receptor subunit rendered the 5-HT\u2083 receptor \u223c70-fold more sensitive to serotonin and produced constitutive activity when combined with the 5-HT_(3B) subunit. Mice homozygous for the mutant allele (5-HT_(3A)^(vs/vs)) had decreased levels of 5-HT_(3A) mRNA. Measurements on sympathetic ganglion cells in these mice showed that whole-cell serotonin responses were reduced, and that the remaining 5-HT\u2083 receptors were hypersensitive. Male 5-HT_(3A)^(vs/vs) mice died at 2-3 months of age, and heterozygous (5-HT_(3A)^(vs/+)) males and homozygous mutant females died at 4-6 months of age from an obstructive uropathy. Both male and female 5-HT_(3A) mutant mice had urinary bladder mucosal and smooth muscle hyperplasia and hypertrophy, whereas male mutant mice had additional prostatic smooth muscle and urethral hyperplasia. 5-HT_(3A) mutant mice had marked voiding dysfunction characterized by a loss of micturition contractions with overflow incontinence. Detrusor strips from 5-HT_(3A)^(vs/vs) mice failed to contract to neurogenic stimulation, despite overall normal responses to a cholinergic agonist, suggestive of altered neuronal signaling in mutant mouse bladders. Consistent with this hypothesis, decreased nerve fiber immunoreactivity was observed in the urinary bladders of 5-HT_(3A)^(vs/vs) compared with 5-HT_(3A) wild-type (5-HT_(3A)^(+/+)) mice. These data suggest that persistent activation of the hypersensitive and constitutively active 5-HT_(3A) receptor in vivo may lead to excitotoxic neuronal cell death and functional changes in the urinary bladder, resulting in bladder hyperdistension, urinary retention, and overflow incontinence.", "date": "2004-06-16", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "24", "number": "24", "publisher": "Society for Neuroscience", "pagerange": "5537-5548", "id_number": "CaltechAUTHORS:20200513-072838651", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200513-072838651", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" } ] }, "doi": "10.1523/jneurosci.5658-03.2004", "pmcid": "PMC6729324", "primary_object": { "basename": "5537.full.pdf", "url": "https://authors.library.caltech.edu/records/q5zer-n0t52/files/5537.full.pdf" }, "resource_type": "article", "pub_year": "2004", "author_list": "Bhattacharya, Anindya; Dang, Hong; et el." }, { "id": "https://authors.library.caltech.edu/records/wvk10-vwn50", "eprint_id": 102795, "eprint_status": "archive", "datestamp": "2023-08-19 13:42:12", "lastmod": "2023-10-20 00:30:38", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dibas-M-I", "name": { "family": "Dibas", "given": "Mohammed I." } }, { "id": "Dahan-D-S", "name": { "family": "Dahan", "given": "David S." } }, { "id": "Leite-J-F", "name": { "family": "Leite", "given": "John F." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Cys-loop receptors: new twists and turns", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2004 Elsevier Ltd. \n\nAvailable online 14 April 2004. \n\nWe thank members of our research groups for comments and the NIH (NS11756 and NS34407) for support.", "abstract": "New hypotheses and predictions have arisen from recent work revealing atomic-scale or near-atomic-scale structures of receptors in the 'Cys-loop' superfamily. How general is the cation\u2013\u03c0 interaction between the natural ligand and a tryptophan residue in the aromatic box, and does this interaction extend to other ligands? What is the pathway from the binding site to gating, and what are the conformational changes during gating and desensitization? Is current flow through intracellular 'portals' in the wall of the channel a general feature? This article discusses these and related questions, emphasizing nicotinic ACh receptors and also discussing data from other members of this superfamily.", "date": "2004-06", "date_type": "published", "publication": "Trends in Neurosciences", "volume": "27", "number": "6", "publisher": "Elsevier", "pagerange": "329-336", "id_number": "CaltechAUTHORS:20200427-094708016", "issn": "0166-2236", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-094708016", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" } ] }, "doi": "10.1016/j.tins.2004.04.002", "resource_type": "article", "pub_year": "2004", "author_list": "Lester, Henry A.; Dibas, Mohammed I.; et el." }, { "id": "https://authors.library.caltech.edu/records/344ar-12g54", "eprint_id": 896, "eprint_status": "archive", "datestamp": "2023-08-22 01:48:41", "lastmod": "2023-10-13 22:01:59", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Clayton-D", "name": { "family": "Clayton", "given": "Daniel" } }, { "id": "Shapovalov-G", "name": { "family": "Shapovalov", "given": "George" } }, { "id": "Maurer-J-A", "name": { "family": "Maurer", "given": "Joshua A." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Kochendoerfer-G-G", "name": { "family": "Kochendoerfer", "given": "Gerd G." } } ] }, "title": "Total chemical synthesis and electrophysiological characterization of mechanosensitive channels from Escherichia coli and Mycobacterium tuberculosis", "ispublished": "pub", "full_text_status": "public", "keywords": "ION-CHANNEL, POTASSIUM CHANNEL, CRYSTAL-STRUCTURE, TERMINAL DOMAIN, PROTEIN, MSCL, RECONSTITUTION, SEMISYNTHESIS, FLUORESCENCE, SELECTIVITY", "note": "\u00a9 2004 by the National Academy of Sciences. \n\nEdited by William F. DeGrado, University of Pennsylvania School of Medicine, Philadelphia, PA, and approved January 14, 2004 (received for review September 5, 2003). \n\nWT recombinant Ec-MscL was provided by Jun-yong Choe and Doug Rees (California Institute of Technology). We thank Erik Rodriguez (California Institute of Technology) for help with studies on the vesicle reconstitution of synthetic Ec-MscL and Christian Becker (Gryphon Therapeutics) for support and discussions. This work was supported by National Institutes of Health Grant GM-062532. \n\nThis paper was submitted directly (Track II) to the PNAS office.\n\nPublished - CLApnas04.pdf
", "abstract": "Total chemical protein synthesis was used to generate multimilligram quantities of the mechanosensitive channel of large conductance from Escherichia coli (Ec-MscL) and Mycobacterium tuberculosis (Tb-MscL). Cysteine residues introduced to allow chemical ligation were masked with cysteine-reactive molecules, resulting in side chain functional groups similar to those of the wild-type protein. Synthetic channel proteins were transferred to 2,2,2-trifluoroethanol and reconstituted into vesicle membranes. Fluorescent imaging of vesicles showed that channel proteins were membrane-localized. Single-channel recordings showed that reconstituted synthetic Ec-MscL has conductance, pressure dependence, and substate distribution similar to those of the recombinant channel. Reconstituted synthetic Tb-MscL also displayed conductance and pressure dependence similar to that of the recombinant protein. Possibilities for the incorporation of unnatural amino acids and biophysical probes, and applications of such synthetic ion channel analogs, are discussed.", "date": "2004-04-06", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "101", "number": "14", "publisher": "National Academy of Sciences", "pagerange": "4764-4769", "id_number": "CaltechAUTHORS:CLApnas04", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:CLApnas04", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-062532" } ] }, "doi": "10.1073/pnas.0305693101", "pmcid": "PMC387322", "primary_object": { "basename": "CLApnas04.pdf", "url": "https://authors.library.caltech.edu/records/344ar-12g54/files/CLApnas04.pdf" }, "resource_type": "article", "pub_year": "2004", "author_list": "Clayton, Daniel; Shapovalov, George; et el." }, { "id": "https://authors.library.caltech.edu/records/ayetn-tvm74", "eprint_id": 64459, "eprint_status": "archive", "datestamp": "2023-08-22 01:15:43", "lastmod": "2023-10-17 21:25:13", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Dickinson-Mary-E", "name": { "family": "Dickinson", "given": "Mary E." } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Jareb-Mark", "name": { "family": "Jareb", "given": "Mark" } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Fraser-S-E", "name": { "family": "Fraser", "given": "Scott E." }, "orcid": "0000-0002-5377-0223" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Assembly of \u03b14\u03b22 Nicotinic Acetylcholine Receptors Assessed with Functional Fluorescently Labeled Subunits: Effects of Localization, Trafficking, and Nicotine-Induced Upregulation in Clonal Mammalian Cells and in Cultured Midbrain Neurons", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotine addiction; nicotinic receptors; receptor assembly; FRET; \u03b14\u03b22; fluorescent protein", "note": "\u00a9 2003 Society for Neuroscience. For the first six months after publication SfN's license will be exclusive. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). \n\nReceived Aug. 13, 2003; revised Oct. 14, 2003; accepted Oct. 16, 2003. \n\nThis work was supported by National Institutes of Health Grants NS11756 and HD37105 and the California Tobacco-Related Disease Research Program (TRDRP). R.N. was supported by a postdoctoral fellowship from the TRDRP (10FT-0174) and the Elizabeth Ross Fellowship. We thank Bruce Cohen and Julian Revie for critical evaluation of this manuscript, Eric Slimko for providing the GluCl\u03b2-YFP GluCl\u03b1-CFP cDNAs, Donghong Ju, Alexa Mundy, Matthew Abramian, and Kai Sung for technical help, Qi Huang for writing some of the data analysis programs, and James Fisher for performing several three-cube FRET experiments.\n\nPublished - 11554.full.pdf
", "abstract": "Fura-2 recording of Ca^(2+) influx was used to show that incubation in 1 \u03bcM nicotine (2-6 d) upregulates several pharmacological components of acetylcholine (ACh) responses in ventral midbrain cultures, including a MLA-resistant, DH\u03b2E-sensitive component that presumably corresponds to \u03b14\u03b22 receptors. To study changes in \u03b14\u03b22 receptor levels and assembly during this upregulation, we incorporated yellow and cyan fluorescent proteins (YFPs and CFPs) into the \u03b14 or \u03b22 M3-M4 intracellular loops, and these subunits were coexpressed in human embryonic kidney (HEK) 293T cells and cultured ventral midbrain neurons. The fluorescent receptors resembled wild-type receptors in maximal responses to ACh, dose-response relations, ACh-induced Ca^(2+) influx, and somatic and dendritic distribution. Transfected midbrain neurons that were exposed to nicotine (1 d) displayed greater levels of fluorescent \u03b14 and \u03b22 nicotinic ACh receptor (nAChR) subunits. As expected from the hetero-multimeric nature of \u03b14\u03b22 receptors, coexpression of the \u03b14-YFP and \u03b22-CFP subunits resulted in robust fluorescence resonance energy transfer (FRET), with a FRET efficiency of 22%. In midbrain neurons, dendritic \u03b14\u03b22 nAChRs displayed greater FRET than receptors inside the soma, and in HEK293T cells, a similar increase was noted for receptors that were translocated to the surface during PKC stimulation. When cultured transfected midbrain neurons were incubated in 1 \u03bcMnicotine, there was increased FRET in the cell body, denoting increased assembly of \u03b14\u03b22 receptors. Thus, changes in \u03b14\u03b22 receptor assembly play a role in the regulation of \u03b14\u03b22 levels and responses in both clonal cell lines and midbrain neurons, and the regulation may result from Ca^(2+)-stimulated pathways.", "date": "2003-12-17", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "23", "number": "37", "publisher": "Society for Neuroscience", "pagerange": "11554-11567", "id_number": "CaltechAUTHORS:20160212-092320526", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160212-092320526", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "HD37105" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "10FT-0174" }, { "agency": "Elizabeth Ross Fellowship" } ] }, "doi": "10.1523/jneurosci.23-37-11554.2003", "pmcid": "PMC6740951", "primary_object": { "basename": "11554.full.pdf", "url": "https://authors.library.caltech.edu/records/ayetn-tvm74/files/11554.full.pdf" }, "resource_type": "article", "pub_year": "2003", "author_list": "Nashmi, Raad; Dickinson, Mary E.; et el." }, { "id": "https://authors.library.caltech.edu/records/njft7-kff72", "eprint_id": 85279, "eprint_status": "archive", "datestamp": "2023-08-19 12:37:14", "lastmod": "2023-10-18 18:03:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Gutman-G-A", "name": { "family": "Gutman", "given": "George A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "International Union of Pharmacology. XLI. Compendium of Voltage-Gated Ion Channels: Potassium Channels", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2003 by the American Society for Pharmacology and Experimental Therapeutics.", "abstract": "This summary article presents an overview of the molecular relationships among the voltage-gated potassium channels and a standard nomenclature for them, which is derived from the IUPHAR Compendium of Voltage-Gated Ion Channels.1 The complete Compendium, including data tables for each member of the potassium channel family can be found at http://www.iuphar-db.org/iuphar-ic/.", "date": "2003-12-01", "date_type": "published", "publication": "Pharmacological Reviews", "volume": "55", "number": "4", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "583-586", "id_number": "CaltechAUTHORS:20180313-125205786", "issn": "0031-6997", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180313-125205786", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1124/pr.55.4.9", "resource_type": "article", "pub_year": "2003", "author_list": "Gutman, George A. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/ged5v-gte18", "eprint_id": 103215, "eprint_status": "archive", "datestamp": "2023-08-22 01:11:37", "lastmod": "2023-10-20 15:47:20", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Owens-J-C", "name": { "family": "Owens", "given": "Jeremy C." } }, { "id": "Balogh-S-A", "name": { "family": "Balogh", "given": "Seth A." } }, { "id": "McClure-Begley-T-D", "name": { "family": "McClure-Begley", "given": "Tristan D." } }, { "id": "Butt-C-M", "name": { "family": "Butt", "given": "Christopher M." } }, { "id": "Labarca-C-G", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Picciotto-M-R", "name": { "family": "Picciotto", "given": "Marina R." } }, { "id": "Wehner-J-M", "name": { "family": "Wehner", "given": "Jeanne M." } }, { "id": "Collins-A-C", "name": { "family": "Collins", "given": "Allan C." } } ] }, "title": "\u03b14\u03b22* Nicotinic Acetylcholine Receptors Modulate the Effects of Ethanol and Nicotine on the Acoustic Startle Response", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Nicotinic Receptor; Nicotine; Acoustic Startle; Genetics", "note": "\u00a9 2003 Research Society on Alcoholism. \n\nReceived July 7, 2003; Accepted September 11, 2003. \n\nSupported by grants from the National Institutes of Health: AA\u201011156, DA015663, and DA\u201000197 (ACC), AA\u201013018 (JMW), AA\u201013350 (JCO), MH\u201016880 (SAB), NS\u201011756 (HAL), and DA\u201013334 (MRP) and a grant from the California TRDRP to HAL.", "abstract": "Background: Ethanol modulates the functional activity of \u03b14\u03b22 neuronal nicotinic cholinergic receptors (nAChR) when measured in vitro, but the potential role of \u03b14\u03b22 nAChRs in regulating behavioral effects of ethanol is unknown. Recently, Tritto et al. (Tritto T, Stitzel JA, Marks MJ, Romm E, Collins AC (2002) Variability in response to nicotine in the LS\u00d7SS RI strains: potential role of polymorphisms in alpha4 and alpha6 nicotinic receptor genes. Pharmacogenetics 12:197\u2013208) reported that a polymorphism (A529T) in the \u03b14 nAChR subunit gene is associated with variability in nicotine's effects on startle in the LS\u00d7SS recombinant inbred (RI) strains. Ethanol also alters the acoustic startle response. Thus, we evaluated the potential role of \u03b14\u03b22 nAChRs in modulating ethanol's effects on acoustic startle.\n\nMethods: The effects of ethanol on acoustic startle were determined in the LS\u00d7SS RI strains. In addition, the effects of ethanol and nicotine were also measured in \u03b14 gain of function and \u03b22 null mutant mice. The \u03b22 mutants do not express the major variant of \u03b14 nAChRs, \u03b14\u03b22.\n\nResults: An association between the \u03b14 A529T polymorphism and ethanol's effects on startle was found in the LS\u00d7SS RI strains; those strains that express the A529 variant of \u03b14 were more sensitive to ethanol\u2010induced depression of startle. The \u03b14 gain of function mutants were more sensitive to the effects of both nicotine and ethanol and the \u03b22 null mutants were less sensitive to both drugs.\n\nConclusions: \u03b14\u03b22\u2010containing nAChRs may play important roles in modulating the effects of both ethanol and nicotine on the acoustic startle response. We suggest that nAChR subunit genes should be evaluated as potential contributors to both alcoholism and tobacco abuse.", "date": "2003-12", "date_type": "published", "publication": "Alcoholism: Clinical and Experimental Research", "volume": "27", "number": "12", "publisher": "Research Society on Alcoholism", "pagerange": "1867-1875", "id_number": "CaltechAUTHORS:20200514-153422419", "issn": "0145-6008", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200514-153422419", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "AA\u201011156" }, { "agency": "NIH", "grant_number": "DA015663" }, { "agency": "NIH", "grant_number": "DA\u201000197" }, { "agency": "NIH", "grant_number": "AA\u201013018" }, { "agency": "NIH", "grant_number": "AA\u201013350" }, { "agency": "NIH", "grant_number": "MH\u201016880" }, { "agency": "NIH", "grant_number": "NS\u201011756" }, { "agency": "NIH", "grant_number": "DA\u201013334" }, { "agency": "California Tobacco-Related Disease Research Program" } ] }, "doi": "10.1097/01.alc.0000102700.72447.0f", "resource_type": "article", "pub_year": "2003", "author_list": "Owens, Jeremy C.; Balogh, Seth A.; et el." }, { "id": "https://authors.library.caltech.edu/records/w3v8x-k9a83", "eprint_id": 904, "eprint_status": "archive", "datestamp": "2023-08-22 01:01:55", "lastmod": "2023-10-13 22:02:12", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Leite-J-F", "name": { "family": "Leite", "given": "John F." } }, { "id": "Blanton-M-P", "name": { "family": "Blanton", "given": "Michael P." } }, { "id": "Shahgholi-M", "name": { "family": "Shahgholi", "given": "Mona" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Conformation-dependent hydrophobic photolabeling of the nicotinic receptor: Electrophysiology-coordinated photochemistry and mass spectrometry", "ispublished": "pub", "full_text_status": "public", "keywords": "QUADRUPOLE ION-TRAP, ACETYLCHOLINE-RECEPTOR, NONCOMPETITIVE ANTAGONIST, ELECTROSPRAY-IONIZATION, VOLTAGE-DEPENDENCE, GABA(A) RECEPTOR, GLYCINE RECEPTOR, AGONIST BINDING, RESTING STATE, CHANNEL", "note": "\u00a9 2003 by the National Academy of Sciences. \n\nEdited by William A. Catterall, University of Washington School of Medicine, Seattle, WA, and approved August 22, 2003 (received for review May 16, 2003). This paper was submitted directly (Track II) to the PNAS office.\n\nThis work was supported by grants from the National Institutes of Health (NS11756, NS34407, NS35786, and NRSA) to J.F.L.\n\nProceedings of the National Academy of Sciences Dec 2003, 100 (26) 16144; DOI: 10.1073/pnas.2637015100\n\nPublished - LEIpnas03.pdf
Erratum - 16144.full.pdf
", "abstract": "We characterized the differential accessibility of the nicotinic acetylcholine receptor all subunit in the open, closed, and desensitized states by using electrophysiology-coordinated photolabeling by several lipophilic probes followed by mass spectrometric analysis. Voltage-clamped oocytes expressing receptors were preincubated with one of the lipophilic probes and were continually exposed to acetylcholine; UV irradiation was applied during 500-ms pulses to + 40 or to -140 mV (which produced closed or approximate to50% open receptors, respectively). In the open state, there was specific probe incorporation within the N-terminal domain at residues that align with the beta8-beta9 loop of the acetylcholine-binding protein. In the closed state, probe incorporation was identified at several sites of the N-terminal domain within the conserved cysteine loop (residues 128-142), the cytoplasmic loop (M3-M4), and M4. The labeling pattern in the M4 region is consistent with previous results, further defining the lipid-exposed face of this transmembrane a-helix. These results show regions within the N-terminal domain that are involved in gating-dependent conformational shifts, confirm that the cysteine loop resides at or near the protein-membrane interface, and show that segments of the M3-M4 loop are near to the lipid bilayer.", "date": "2003-10-28", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "100", "number": "22", "publisher": "National Academy of Sciences", "pagerange": "13054-13059", "id_number": "CaltechAUTHORS:LEIpnas03", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LEIpnas03", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "NS35786" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1073/pnas.2133028100", "pmcid": "PMC240743", "primary_object": { "basename": "16144.full.pdf", "url": "https://authors.library.caltech.edu/records/w3v8x-k9a83/files/16144.full.pdf" }, "related_objects": [ { "basename": "LEIpnas03.pdf", "url": "https://authors.library.caltech.edu/records/w3v8x-k9a83/files/LEIpnas03.pdf" } ], "resource_type": "article", "pub_year": "2003", "author_list": "Leite, John F.; Blanton, Michael P.; et el." }, { "id": "https://authors.library.caltech.edu/records/q98db-jdc54", "eprint_id": 102819, "eprint_status": "archive", "datestamp": "2023-09-15 06:29:41", "lastmod": "2023-10-23 21:26:46", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Storch-A", "name": { "family": "Storch", "given": "Alexander" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Boehm-B-O", "name": { "family": "Boehm", "given": "Bernhard O." } }, { "id": "Schwarz-J", "name": { "family": "Schwarz", "given": "Johannes" } } ] }, "title": "Functional characterization of dopaminergic neurons derived from rodent mesencephalic progenitor cells", "ispublished": "pub", "full_text_status": "public", "keywords": "Parkinson's disease; Neuroregeneration; Dopaminergic neurons; Transplantation; Neural progenitor cell; Patch\u2013clamp recordings", "note": "\u00a9 2003 Elsevier B.V. \n\nReceived 6 September 2002, Revised 1 April 2003, Accepted 28 June 2003, Available online 26 September 2003.", "abstract": "Neural progenitor cells existing in the developing and adult brain retain the capacity to self renew and to produce the major cell types of the brain opening new avenues for restorative therapy of neuropsychiatric disorders. These cells can be grown in vitro while retaining the potential to differentiate into nervous tissue. A primary target for neurorestoration is Parkinson's disease, characterized by a continuous loss of the dopaminergic neurons in the substantia nigra pars compacta leading to dopamine depletion in the striatum and subsequent clinical symptoms including bradykinesia, rigidity and tremor. We established a protocol for long-term expansion and dopaminergic differentiation of rodent and human mesencephalic neural progenitor cells. Here we perform functional studies using both biochemical and electrophysiological techniques on dopaminergic neurons derived from rodent mesencephalic progenitor cells labeled with tyrosine hydroxylase (TH) gene promotor-driven expression of enhanced green fluorescence protein (EGFP). Thus, we demonstrate that these cells produce and release dopamine, express voltage-gated potassium and sodium currents, and fire action potentials. Furthermore, we detect a slowly activating hyperpolarization-activated inward cation current (Ih), which is specific for dopaminergic neurons among present midbrain neurons. Our results demonstrate that differentiated mesencephalic progenitors exhibit some major morphological and functional characteristics of dopaminergic neurons. Therefore, these neural progenitor cells might serve as a useful source of dopaminergic neurons for studying the development and degeneration of these cells and may further serve as a continuous, on-demand source of cells for therapeutic transplantation in Parkinson's disease.", "date": "2003-10", "date_type": "published", "publication": "Journal of Chemical Neuroanatomy", "volume": "26", "number": "2", "publisher": "Elsevier", "pagerange": "133-142", "id_number": "CaltechAUTHORS:20200427-151732404", "issn": "0891-0618", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-151732404", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/s0891-0618(03)00067-x", "resource_type": "article", "pub_year": "2003", "author_list": "Storch, Alexander; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/2dpmv-kze25", "eprint_id": 102868, "eprint_status": "archive", "datestamp": "2023-09-15 06:29:51", "lastmod": "2023-10-23 21:26:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Jensen-K", "name": { "family": "Jensen", "given": "Kimmo" } }, { "id": "Chiu-Chi-Sung", "name": { "family": "Chiu", "given": "Chi-Sung" } }, { "id": "Sokolova-I-V", "name": { "family": "Sokolova", "given": "Irina" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Mody-I", "name": { "family": "Mody", "given": "Istvan" } } ] }, "title": "GABA Transporter-1 (GAT1)-Deficient Mice: Differential Tonic Activation of GABA_A Versus GABA_B Receptors in the Hippocampus", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2003 by the American Physiological Society. \n\nSubmitted 13 March 2003; accepted in final form 9 June 2003. \n\nThis work was supported by National Institutes of Health Grants DA-09121, DA-010509, NS-11756, MH-49176, MH-61468, NS-30549, DA-14947, National Science Foundation Grant 0119493, and a Della Martin Fellowship to C.-S. Chiu. \n\nThe costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. \n\nK. Jensen and C.-S. Chiu contributed equally to this work.", "abstract": "After its release from interneurons in the CNS, the major inhibitory neurotransmitter GABA is taken up by GABA transporters (GATs). The predominant neuronal GABA transporter GAT1 is localized in GABAergic axons and nerve terminals, where it is thought to influence GABAergic synaptic transmission, but the details of this regulation are unclear. To address this issue, we have generated a strain of GAT1-deficient mice. We observed a large increase in a tonic postsynaptic hippocampal GABA_A receptor-mediated conductance. There was little or no change in the waveform or amplitude of spontaneous inhibitory postsynaptic currents (IPSCs) or miniature IPSCs. In contrast, the frequency of quantal GABA release was one-third of wild type (WT), although the densities of GABA_A receptors, GABA_B receptors, glutamic acid decarboxylase 65 kDa, and vesicular GAT were unaltered. The GAT1-deficient mice lacked a presynaptic GABA_B receptor tone, present in WT mice, which reduces the frequency of spontaneous IPSCs. We conclude that GAT1 deficiency leads to enhanced extracellular GABA levels resulting in an overactivation of GABA_A receptors responsible for a postsynaptic tonic conductance. Chronically elevated GABA levels also downregulate phasic GABA release and reduce presynaptic signaling via GABA_B receptors thus causing an enhanced tonic and a diminished phasic inhibition.", "date": "2003-10", "date_type": "published", "publication": "Journal of Neurophysiology", "volume": "90", "number": "4", "publisher": "American Physiological Society", "pagerange": "2690-2701", "id_number": "CaltechAUTHORS:20200428-095147178", "issn": "0022-3077", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200428-095147178", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "DA-010509" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "MH-61468" }, { "agency": "NIH", "grant_number": "NS-30549" }, { "agency": "NIH", "grant_number": "DA-14947" }, { "agency": "NSF", "grant_number": "CBET-0119493" }, { "agency": "Della Martin Foundation" } ] }, "doi": "10.1152/jn.00240.2003", "resource_type": "article", "pub_year": "2003", "author_list": "Jensen, Kimmo; Chiu, Chi-Sung; et el." }, { "id": "https://authors.library.caltech.edu/records/6gtv2-t0w14", "eprint_id": 102839, "eprint_status": "archive", "datestamp": "2023-08-19 11:58:00", "lastmod": "2023-10-20 00:32:44", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Fonck-C", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "Tapper-A-R", "name": { "family": "Tapper", "given": "Andrew R." } }, { "id": "McKinney-S-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Damaj-M-I", "name": { "family": "Damaj", "given": "M. Imad" } }, { "id": "Balogh-S-A", "name": { "family": "Balogh", "given": "Seth" } }, { "id": "Owens-J", "name": { "family": "Owens", "given": "Jeremy" } }, { "id": "Wehner-J-M", "name": { "family": "Wehner", "given": "Jeanne M." } }, { "id": "Collins-A-C", "name": { "family": "Collins", "given": "Allan C." } }, { "id": "Labarca-C-G", "name": { "family": "Labarca", "given": "Cesar" } } ] }, "title": "Hypersensitive knockin mouse strains identify receptors and pathways for nicotine action", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2003 Current Drugs.", "abstract": "Two series of knockin mouse strains have been constructed with point mutations that result in hypersensitive neuronal nicotinic acetylcholine receptors containing \u03b1\u2084- or \u03b1\u2087-subunits. The full expression of the stronger alleles produces neonatal excitotoxic lethality; however, mice with attenuated expression or milder alleles are viable, and display a range of hypersensitive responses to nicotine. To date, measurements have been made on nicotine-induced seizures, Straub tail, hypothermia, antinociception, electroencephalograms and cellular electrophysiological responses. These strains are helping to define the occurrence of these important receptor subtypes, and their role in the acute and chronic actions of nicotine. The hypersensitive strains may be useful for the development of nicotinic drug therapy.", "date": "2003-09", "date_type": "published", "publication": "Current opinion in drug discovery & development", "volume": "6", "number": "5", "publisher": "Current Drugs", "pagerange": "633-639", "id_number": "CaltechAUTHORS:20200428-093705406", "issn": "1367-6733", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200428-093705406", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "resource_type": "article", "pub_year": "2003", "author_list": "Lester, Henry A.; Fonck, Carlos; et el." }, { "id": "https://authors.library.caltech.edu/records/rqnjd-b7155", "eprint_id": 102623, "eprint_status": "archive", "datestamp": "2023-08-19 11:52:17", "lastmod": "2023-10-20 00:21:13", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Leite-J-F", "name": { "family": "Leite", "given": "John F." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Shahgholi-M", "name": { "family": "Shahgholi", "given": "Mona" } } ] }, "title": "Investigation of apparent mass deviations in electrospray ionization tandem mass spectrometry of a benzophenone-labeled peptide", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2003 John Wiley & Sons. \n\nReceived 8 April 2003; Revised 22 May 2003; Accepted 22 May 2003. \n\nThe authors gratefully acknowledge fruitful discussions with J. Callahan and R. Vachet. This work was supported by a National Institute of Health National Research Service Award fellowship.", "abstract": "In a previous study utilizing benzophenone\u2010based topological probes to study conformationally dependent changes in mouse muscle nicotinic acetylcholine receptor (nAChR) topology, electrospray ionization tandem mass spectrometric (ESI\u2010MS/MS) analysis led to a consistent \u22122.0\u2009Da mass deviation from expected values. In the present study a synthetic peptide, corresponding to nAChR \u03b11 subunit residues 130\u2013139, was photolabeled. MS/MS analysis of this peptide using an ion trap confirmed the previously observed mass deviation, associated only with fragment ions that contain the incorporated benzophenone moiety. Analysis of peak profiles for the photolabeled ions does not indicate the typical 'peak fronting' that produces a mass shift when labile ions are prematurely ejected from the ion trap. Rather, hydrogen/deuterium (H/D) exchange experiments support the hypothesis that a chemical rearrangement involving phenyl migration and ketone formation has formed an unexpected oxidized peptide, with molecular mass 2\u2009Da less than that expected, that is isolated for collision\u2010induced dissociation in the ion trap together with the predicted precursor due to the broad ion isolation window specified.", "date": "2003-08-15", "date_type": "published", "publication": "Rapid Communications in Mass Spectrometry", "volume": "17", "number": "15", "publisher": "Wiley", "pagerange": "1677-1684", "id_number": "CaltechAUTHORS:20200417-142717107", "issn": "0951-4198", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200417-142717107", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1002/rcm.1103", "resource_type": "article", "pub_year": "2003", "author_list": "Leite, John F.; Dougherty, Dennis A.; et el." }, { "id": "https://authors.library.caltech.edu/records/q0tmp-sz344", "eprint_id": 102869, "eprint_status": "archive", "datestamp": "2023-08-19 11:43:26", "lastmod": "2023-10-20 00:34:52", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Rodrigues-Pinguet-Nivalda-O", "name": { "family": "Rodrigues-Pinguet", "given": "Nivalda" } }, { "id": "Jia-Li", "name": { "family": "Jia", "given": "Li" } }, { "id": "Li-Maureen", "name": { "family": "Li", "given": "Maureen" } }, { "id": "Figl-Antonio", "name": { "family": "Figl", "given": "Antonio" } }, { "id": "Klaassen-Alwin", "name": { "family": "Klaassen", "given": "Alwin" } }, { "id": "Truong-Anthony", "name": { "family": "Truong", "given": "Anthony" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Cohen-Bruce-N", "name": { "family": "Cohen", "given": "Bruce N." } } ] }, "title": "Five ADNFLE Mutations Reduce the Ca\u00b2\u207a Dependence of the Mammalian \u03b14\u03b22 Acetylcholine Response", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2003 The Physiological Society. \n\nReceived 10 February 2003; accepted after revision 15 April 2003; first published online 16 May 2003) \n\nThis research was supported by the California Tobacco\u2010Related Disease Research Program (6KT\u20100208), the Epilepsy Foundation (EFA/COHEN\u201001) and the NIH (NS43800, NS11756). We thank Kira Kostenka for help with the oocytes and Cesar Labarca, Purnima Deshpande, Carlos Fonck, Raad Nashmi, Andrew Tapper, Johannes Schwarz and Jim Boulter for comments.", "abstract": "Five nicotinic acetylcholine receptor (nAChR) mutations are currently linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE). The similarity of their clinical symptoms suggests that a common functional anomaly of the mutations underlies ADNFLE seizures. To identify this anomaly, we constructed rat orthologues (S252F, +L264, S256L, V262L, V262M) of the human ADNFLE mutations, expressed them in Xenopus oocytes with the appropriate wild\u2010type (WT) subunit (\u03b14 or \u03b22), and studied the Ca\u00b2\u207a dependence of their ACh responses. All the mutations significantly reduced 2 mM Ca\u00b2\u207a\u2010induced increases in the 30 \u03bcM ACh response (P < 0.05). Consistent with a dominant mode of inheritance, this reduction persisted in oocytes injected with a 1:1 mixture of mutant and WT cRNA. BAPTA injections showed that the reduction was not due to a decrease in the secondary activation of Ca\u00b2\u207a\u2010activated Cl\u207b currents. The S256L mutation also abolished 2 mM Ba\u00b2\u207a potentiation of the ACh response. The S256L, V262L and V262M mutations had complex effects on the ACh concentration\u2010response relationship but all three mutations shifted the concentration\u2010response relationship to the left at [ACh]\u2a7e 30 \u03bcM. Co\u2010expression of the V262M mutation with a mutation (E180Q) that abolished Ca\u00b2\u207a potentiation resulted in 2 mM Ca\u00b2\u207a block, rather than potentiation, of the 30 \u03bcM ACh response, suggesting that the ADNFLE mutations reduce Ca\u00b2\u207a potentiation by enhancing Ca\u00b2\u207a block of the \u03b14\u03b22 nAChR. Ca\u00b2\u207a modulation may prevent presynaptic \u03b14\u03b22 nAChRs from overstimulating glutamate release at central excitatory synapses during bouts of synchronous, repetitive activity. Reducing the Ca\u00b2\u207a dependence of the ACh response could trigger seizures by increasing \u03b14\u03b22\u2010mediated glutamate release during such bouts.", "date": "2003-07", "date_type": "published", "publication": "Journal of Physiology", "volume": "550", "number": "1", "publisher": "Physiological Society", "pagerange": "11-26", "id_number": "CaltechAUTHORS:20200428-095147285", "issn": "0022-3751", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200428-095147285", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "6KT\u20100208" }, { "agency": "Epilepsy Foundation of America", "grant_number": "EFA/COHEN\u201001" }, { "agency": "NIH", "grant_number": "NS43800" }, { "agency": "NIH", "grant_number": "NS11756" } ] }, "doi": "10.1113/jphysiol.2003.036681", "pmcid": "PMC2343021", "resource_type": "article", "pub_year": "2003", "author_list": "Rodrigues-Pinguet, Nivalda; Jia, Li; et el." }, { "id": "https://authors.library.caltech.edu/records/84dmg-8pw63", "eprint_id": 103235, "eprint_status": "archive", "datestamp": "2023-08-22 00:41:29", "lastmod": "2023-10-20 15:48:50", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Yu-Tzu-ping", "name": { "family": "Yu", "given": "Tzu-ping" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Requirement of a critical period of GABAergic receptor blockade for induction of a cAMP-mediated long-term depression at CA3-CA1 synapses", "ispublished": "pub", "full_text_status": "restricted", "keywords": "hippocampus; slice culture; LTP; protein kinase A; protein phosphatase; CREB", "note": "\u00a9 2003 Wiley\u2010Liss, Inc. \n\nReceived 22 October 2002; Accepted 25 February 2003. \n\nContract grant sponsor: NIH; Contract grant numbers: MH-49176 and NS-11756. \n\nWe thank Dong-hung Ju for technical assistance in immunostaining experiments, Melvin Simon for providing the Nikon microscope, Chi-sung Chiu for discussion, and Erin Schuman for critical reading of the manuscript and helpful comments.", "abstract": "Previous reports show that bath application of the adenosine 3\u2032 : 5\u2032\u2010cyclic monophosphate (cAMP) analog, Sp\u2010cAMPS, induces a protein kinase A (PKA)\u2010dependent and protein synthesis\u2010dependent long\u2010term potentiation (LTP) at hippocampal CA3\u2010CA1 synapses. Recently, we reported a novel form of long\u2010term depression (LTD) induced by concurrent application of Sp\u2010cAMPS and picrotoxin, the \u03b3\u2010aminobutyric acid type A (GABA_A) receptor antagonist. In the present study, we further investigated the mechanisms underlying such cAMP\u2010mediated LTD. Synaptically connected CA3 and CA1 cells of hippocampal slice cultures were impaled by sharp electrodes. Excitatory postsynaptic potentials recorded from a CA1 pyramidal cell were evoked by single action potentials in a CA3 cell. Picrotoxin was applied to slices at various time points after Sp\u2010cAMPS was perfused. We found that Sp\u2010cAMPS\u2010induced potentiation could be converted to depression when picrotoxin was applied within 30 min after perfusion of Sp\u2010cAMPS. Picrotoxin applied 1 h after perfusion of Sp\u2010cAMPS had no effect on Sp\u2010cAMPS\u2010induced synaptic potentiation. Once LTP was induced by Sp\u2010cAMPS and expressed for 1 h, the subsequent application of Sp\u2010cAMPS and picrotoxin produced no new changes in synaptic strength. Also, once LTD was induced and expressed for 1 h, subsequent Sp\u2010cAMPS produced no new changes in synaptic strength. These findings suggest that a synapse is committed irreversibly to cAMP\u2010mediated LTP or LTD during a critical period and that later signals cannot interconvert these two fates.", "date": "2003-07", "date_type": "published", "publication": "Synapse", "volume": "49", "number": "1", "publisher": "Wiley", "pagerange": "12-19", "id_number": "CaltechAUTHORS:20200515-112555466", "issn": "0887-4476", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200515-112555466", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1002/syn.10207", "resource_type": "article", "pub_year": "2003", "author_list": "Yu, Tzu-ping; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/t669p-1xy37", "eprint_id": 103237, "eprint_status": "archive", "datestamp": "2023-08-19 11:38:12", "lastmod": "2023-10-20 15:49:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Rahman-Z", "name": { "family": "Rahman", "given": "Zia" } }, { "id": "Schwarz-J", "name": { "family": "Schwarz", "given": "Johannes" } }, { "id": "Gold-S-J", "name": { "family": "Gold", "given": "Stephen J." } }, { "id": "Zachariou-V", "name": { "family": "Zachariou", "given": "Venetia" } }, { "id": "Wein-M-N", "name": { "family": "Wein", "given": "Marc N." } }, { "id": "Choi-Kwang-Ho", "name": { "family": "Choi", "given": "Kwang-Ho" } }, { "id": "Kovoor-A", "name": { "family": "Kovoor", "given": "Abraham" } }, { "id": "Chen-Ching-Kang", "name": { "family": "Chen", "given": "Ching-Kang" } }, { "id": "DiLeone-R-J", "name": { "family": "DiLeone", "given": "Ralph J." } }, { "id": "Schwarz-S-C", "name": { "family": "Schwarz", "given": "Sigrid C." } }, { "id": "Selley-D-E", "name": { "family": "Selley", "given": "Dana E." } }, { "id": "Sim-Selley-L-J", "name": { "family": "Sim-Selley", "given": "Laura J." } }, { "id": "Barrot-M", "name": { "family": "Barrot", "given": "Michel" } }, { "id": "Luedtke-R-R", "name": { "family": "Luedtke", "given": "Robert R." } }, { "id": "Self-D", "name": { "family": "Self", "given": "David" } }, { "id": "Neve-R-L", "name": { "family": "Neve", "given": "Rachael L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Simon-M-I", "name": { "family": "Simon", "given": "Melvin I." } }, { "id": "Nestler-E-J", "name": { "family": "Nestler", "given": "Eric J." } } ] }, "title": "RGS9 Modulates Dopamine Signaling in the Basal Ganglia", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2003 Cell Press. \n\nReceived 18 October 2002, Revised 3 March 2003, Accepted 25 April 2003, Available online 17 April 2004. \n\nWe wish to thank Dr. William Carlezon, Jr., for helpful discussions; Drs. Vadim Arshavsky, W.F. Simonds, and Andrejs Krummins for antibodies; Cathy Steffen for animal care; and Mike Cassidy, Christina Colby, and Lee Schlesinger for excellent technical support. This work was supported by grants from the National Institute on Drug Abuse (DA08227, DA05274), National Institute of Mental Health (MH66172), National Institute of General Medical Services (GM29836), the Plum Foundation, and by a gift from Ruben Mettler.", "abstract": "Regulators of G protein signaling (RGS) modulate heterotrimeric G proteins in part by serving as GTPase-activating proteins for G\u03b1 subunits. We examined a role for RGS9-2, an RGS subtype highly enriched in striatum, in modulating dopamine D2 receptor function. Viral-mediated overexpression of RGS9-2 in rat nucleus accumbens (ventral striatum) reduced locomotor responses to cocaine (an indirect dopamine agonist) and to D2 but not to D1 receptor agonists. Conversely, RGS9 knockout mice showed heightened locomotor and rewarding responses to cocaine and related psychostimulants. In vitro expression of RGS9-2 in Xenopus oocytes accelerated the off-kinetics of D2 receptor-induced GIRK currents, consistent with the in vivo data. Finally, chronic cocaine exposure increased RGS9-2 levels in nucleus accumbens. Together, these data demonstrate a functional interaction between RGS9-2 and D2 receptor signaling and the behavioral actions of psychostimulants and suggest that psychostimulant induction of RGS9-2 represents a compensatory adaptation that diminishes drug responsiveness.", "date": "2003-06-19", "date_type": "published", "publication": "Neuron", "volume": "38", "number": "6", "publisher": "Cell Press", "pagerange": "941-952", "id_number": "CaltechAUTHORS:20200515-125021148", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200515-125021148", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA08227" }, { "agency": "NIH", "grant_number": "DA05274" }, { "agency": "NIH", "grant_number": "MH66172" }, { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "Plum Foundation" }, { "agency": "Ruben Mettler" } ] }, "doi": "10.1016/s0896-6273(03)00321-0", "resource_type": "article", "pub_year": "2003", "author_list": "Rahman, Zia; Schwarz, Johannes; et el." }, { "id": "https://authors.library.caltech.edu/records/d9mv6-7r133", "eprint_id": 77328, "eprint_status": "archive", "datestamp": "2023-08-19 11:36:52", "lastmod": "2023-10-25 22:05:09", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Mu-Ting-Wei", "name": { "family": "Mu", "given": "Ting-Wei" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Different Binding Orientations for the Same Agonist at Homologous Receptors: A Lock and Key or a Simple Wedge?", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2003 American Chemical Society. \n\nReceived February 21, 2003. Publication Date (Web): May 16, 2003. \n\nThis work was support by the NIH (NS-34407 and NS-11756). We thank Darren Beene for helpful discussion.\n\nSupplemental Material - ja0348086si20030415_012813.pdf
", "abstract": "Using unnatural amino acid mutagenesis, the binding site for serotonin at the novel Caenorhabditis elegans receptor MOD-1 has been probed. As with the closely related serotonin receptor 5-HT_3, MOD-1 makes use of a strong cation\u2212\u03c0 interaction between the ammonium of serotonin and the indole side chain of a tryptophan. However, the specific Trp used by MOD-1 is different from that used for 5-HT_3 (and the nAChR), aligning with a residue more than 40 amino acids distant in sequence space and on a different \"loop\" of the agonist binding site. This suggests a significant rearrangement of the ligand on binding these two closely related receptors. It is suggested that, unlike enzymes, receptors and other signaling molecules may need only to deliver an agonist to a general binding region, rather than establishing precise drug\u2212receptor interactions.", "date": "2003-06-11", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "125", "number": "23", "publisher": "American Chemical Society", "pagerange": "6850-6851", "id_number": "CaltechAUTHORS:20170510-080843497", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170510-080843497", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1021/ja0348086", "primary_object": { "basename": "ja0348086si20030415_012813.pdf", "url": "https://authors.library.caltech.edu/records/d9mv6-7r133/files/ja0348086si20030415_012813.pdf" }, "resource_type": "article", "pub_year": "2003", "author_list": "Mu, Ting-Wei; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/2w9td-9k606", "eprint_id": 102813, "eprint_status": "archive", "datestamp": "2023-08-19 11:35:18", "lastmod": "2023-10-20 00:31:25", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Monahan-S-L", "name": { "family": "Monahan", "given": "Sarah L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Site-Specific Incorporation of Unnatural Amino Acids into Receptors Expressed in Mammalian Cells", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2003 Cell Press. Published by Elsevier Ltd. \n\nReceived 2 April 2003, Revised 8 May 2003, Accepted 13 May 2003, Available online 24 June 2003. \n\nWe thank Jack Horne, Gabriel Brandt, and Raad Nashmi for their helpful advice and Mike Walsh (electronic shop) and Herb Adams (machine shop) for their assistance. Supported by NIH (NS-34407, NS-11756, GM-29836). S.L.M. held an NSERC (PGS-B) scholarship and was a fellow of an NIH Predoctoral Trainee Grant (GM-08501).", "abstract": "We describe an approach to achieve unnatural amino acid incorporation into channels and receptors expressed in mammalian cells. We show that microelectroporation provides a general method to deliver DNA, mRNA, and tRNA simultaneously. In both CHO cells and cultured neurons, microelectroporation efficiently delivers an in vitro transcribed, serine amber suppressor tRNA, leading to nonsense suppression in a mutant EGFP gene. In CHO cells, both natural and unnatural amino acids chemically appended to a suppressor tRNA are site specifically incorporated into the nicotinic acetylcholine receptor (nAChR). Electrophysiology confirms the expected functional consequences of the unnatural residue. The microelectroporation strategy described here is more general, less tedious, and less damaging to mammalian neuronal and nonneuronal cells than previous approaches to nonsense suppression in small cells and provides the first example of unnatural amino acid incorporation in mammalian cells using chemically aminoacylated tRNA.", "date": "2003-06", "date_type": "published", "publication": "Chemistry and Biology", "volume": "10", "number": "6", "publisher": "Elsevier", "pagerange": "573-580", "id_number": "CaltechAUTHORS:20200427-134043259", "issn": "1074-5521", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-134043259", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM-08501" } ] }, "doi": "10.1016/s1074-5521(03)00124-8", "resource_type": "article", "pub_year": "2003", "author_list": "Monahan, Sarah L.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/1wtdh-mds38", "eprint_id": 102806, "eprint_status": "archive", "datestamp": "2023-08-19 11:35:10", "lastmod": "2023-10-20 00:31:14", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Beene-D-L", "name": { "family": "Beene", "given": "Darren L." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Unnatural amino acid mutagenesis in mapping ion channel function", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2003 Elsevier Science Ltd. \n\nAvailable online 11 June 2003.", "abstract": "Unnatural amino acid mutagenesis makes possible the site-specific incorporation of synthetic amino acids, enabling detailed structure\u2013function studies as well as the incorporation of biophysical probes. This method has been adapted for use with heterologous expression in Xenopus oocytes, allowing experiments on ion channels.", "date": "2003-06", "date_type": "published", "publication": "Current Opinion in Neurobiology", "volume": "13", "number": "3", "publisher": "Elsevier", "pagerange": "264-270", "id_number": "CaltechAUTHORS:20200427-123619296", "issn": "0959-4388", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200427-123619296", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/s0959-4388(03)00068-0", "resource_type": "article", "pub_year": "2003", "author_list": "Beene, Darren L.; Dougherty, Dennis A.; et el." }, { "id": "https://authors.library.caltech.edu/records/bnx2n-3wh18", "eprint_id": 98898, "eprint_status": "archive", "datestamp": "2023-08-22 00:23:16", "lastmod": "2023-10-18 17:43:14", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Fonck-Carlos", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "Nashmi-Raad", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Deshpande-Purnima", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Damaj-M-Imad", "name": { "family": "Damaj", "given": "M. Imad" } }, { "id": "Marks-Michael-J", "name": { "family": "Marks", "given": "Michael J." } }, { "id": "Riedel-Anett", "name": { "family": "Riedel", "given": "Anett" } }, { "id": "Schwarz-Johannes", "name": { "family": "Schwarz", "given": "Johannes" } }, { "id": "Collins-Allan-C", "name": { "family": "Collins", "given": "Allan C." } }, { "id": "Labarca-Cesar-G", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Increased Sensitivity to Agonist-Induced Seizures, Straub Tail, and Hippocampal Theta Rhythm in Knock-In Mice Carrying Hypersensitive \u03b14 Nicotinic Receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotinic receptor; ADNFLE; seizure; epilepsy; cholinergic; gain of function; knock-in; mouse", "note": "\u00a9 2003 Society for Neuroscience. \n\nReceived Sept. 17, 2002; revised Dec. 24, 2002; accepted Jan. 13, 2003. \n\nThis work was supported by the Tobacco-Related Disease Research Program; by the Keck Foundation; by National Institutes of Health Grants NS-117656, MH-49176, DA-10156, and DA-11836; and by a National Research Service Award to C.F. We thank Jim Boulter, Bruce Cohen, Bonnie Davis, Ken Davis, Sharon Grady, Margaret Jacobs, Yuan Liu, and Jeanne Wehner for discussion; Sarah McCallum for experiments on GABA release; Istvan Mody and Enric Claverol for teaching us electroencephalography; and Bronagh Glaser for administrative assistance.\n\nPublished - 2582.full.pdf
", "abstract": "We studied a strain of exon replacement mice (\"L9\u2032S knock-in\") whose \u03b14 nicotinic receptor subunits have a leucine to serine mutation in the M2 region, 9\u2032 position (Labarca et al., 2001); this mutation renders \u03b14-containing receptors hypersensitive to agonists. Nicotine induced seizures at concentrations (1 mg/kg) approximately eight times lower in L9\u2032S than in wild-type (WT) littermates. At these concentrations, L9\u2032S but not WT showed increases in EEG amplitude and theta rhythm. L9\u2032S mice also showed higher seizure sensitivity to the nicotinic agonist epibatidine, but not to the GABA_Areceptor blocker and proconvulsant bicuculline. Dorsiflexion of the tail (Straub tail) was the most sensitive nicotine effect found in L9\u2032S mice (0.1 mg/kg). The L9\u2032S mice were hypersensitive to galanthamine- and tacrine-induced seizures and Straub tails. There were no apparent neuroanatomical differences between L9\u2032S and WT mice in several brain regions. [125I]Epibatidine binding to brain membranes showed that the mutant allele was expressed at \u223c25% of WT levels, presumably because of the presence of a neomycin selection cassette in a nearby intron. ^(86)Rb efflux experiments on brain synaptosomes showed an increased fraction of function at low agonist concentrations in L9\u2032S mice. These data support the possible involvement of gain-of-function \u03b14 receptors in autosomal dominant nocturnal frontal-lobe epilepsy.", "date": "2003-04-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "23", "number": "7", "publisher": "Society for Neuroscience", "pagerange": "2582-2590", "id_number": "CaltechAUTHORS:20190927-090347351", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190927-090347351", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "W. M. Keck Foundation" }, { "agency": "NIH", "grant_number": "NS-117656" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "DA-10156" }, { "agency": "NIH", "grant_number": "DA-11836" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1523/jneurosci.23-07-02582.2003", "pmcid": "PMC6742094", "primary_object": { "basename": "2582.full.pdf", "url": "https://authors.library.caltech.edu/records/bnx2n-3wh18/files/2582.full.pdf" }, "resource_type": "article", "pub_year": "2003", "author_list": "Fonck, Carlos; Nashmi, Raad; et el." }, { "id": "https://authors.library.caltech.edu/records/kfn6b-qe408", "eprint_id": 52788, "eprint_status": "archive", "datestamp": "2023-08-19 11:15:26", "lastmod": "2023-10-18 21:35:15", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Shapovalov-G", "name": { "family": "Shapovalov", "given": "George" } }, { "id": "Bass-R-B", "name": { "family": "Bass", "given": "Randal" } }, { "id": "Rees-D-C", "name": { "family": "Rees", "given": "Douglas C." }, "orcid": "0000-0003-4073-1185" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Open-State Disulfide Crosslinking between Mycobacterium tuberculosis Mechanosensitive Channel Subunits", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2003 by the Biophysical Society. \n\nSubmitted August 30, 2002, and accepted for publication December 9,\n2002. \n\nWe thank Sergei Sukharev and Paul Blount for introducing us to MscL electrophysiology, Josh Maurer for mutants and cells, and Ido Braslavsky, Donald Elmore, Steve Quake, Gerd Kochendoerfer, and Dennis Dougherty for valuable discussion.\n\nThis research was supported by a grant from the National Institutes of Health (GM-062532), by a Burroughs-Wellcome Fund Computation and Molecular Biology Fellowship to George Shapovalov, and by an National Research Service Award to Randal Bass (GM-020705).\n\nPublished - shapovalov_2003.pdf
", "abstract": "The mechanosensitive channel of large conductance from Mycobacterium tuberculosis (Tb-MscL) was subjected to cysteine-scanning mutagenesis at several residues in the M1 region. The V15C channel displayed disulfide crosslinking in air, but not in the presence of 100 mM \u03b2-mercaptoethanol. In single-channel experiments, the V15C channel was more sensitive to tension than was wild-type Tb-MscL. In air, Tb-MscL V15C occasionally displayed signature-events: at constant tension, there was first a sojourn in the highest conductance open state, then a series of transitions to substates. During a signature-event, these transitions do not appear to be reversible. Some sojourns in the lower conductance states lasted for \u2265100 s. These signature-events were abolished by 100 mM \u03b2-mercaptoethanol and did not occur in a cysteineless gain-of-function mutant, suggesting that the signature-events represent disulfide crosslinking between channel subunits. We conclude that the crosslinking occurs during an open state during asymmetric sojourns that bring the \u03b1-carbons of adjacent 15C side chains within 3.6\u20136.8 \u00c5. Such asymmetric structures must be considered in models of TB-MscL gating.", "date": "2003-04", "date_type": "published", "publication": "Biophysical Journal", "volume": "84", "number": "4", "publisher": "Biophysical Society", "pagerange": "2357-2365", "id_number": "CaltechAUTHORS:20141212-145345633", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20141212-145345633", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-062532" }, { "agency": "Burroughs-Wellcome Fund" }, { "agency": "NIH", "grant_number": "GM-020705" } ] }, "doi": "10.1016/S0006-3495(03)75041-3", "pmcid": "PMC1302802", "primary_object": { "basename": "shapovalov_2003.pdf", "url": "https://authors.library.caltech.edu/records/kfn6b-qe408/files/shapovalov_2003.pdf" }, "resource_type": "article", "pub_year": "2003", "author_list": "Shapovalov, George; Bass, Randal; et el." }, { "id": "https://authors.library.caltech.edu/records/c2xhj-ek245", "eprint_id": 104091, "eprint_status": "archive", "datestamp": "2023-08-22 00:20:45", "lastmod": "2023-10-20 19:04:31", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Slimko-E-M", "name": { "family": "Slimko", "given": "Eric M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Codon optimization of Caenorhabditis elegans GluCl ion channel genes for mammalian cells dramatically improves expression levels", "ispublished": "pub", "full_text_status": "restricted", "keywords": "GluCl; Codon optimization; Fusion proteins; Electrophysiology; Excitability; EYFP; ECFP", "note": "\u00a9 2003 Elsevier. \n\nReceived 18 October 2002; received in revised form 11 December 2002; accepted 11 December 2002. \n\nThis work was supported by National Institutes of Health (NS 11756 and MH 49176), by the Plum Foundation and by the William T. Gimbel Discovery fund in Neuroscience. We thank David Anderson, Michael Fanselow and Christof Koch for discussion and Sheri McKinney for help with cultures.", "abstract": "Organisms use synonymous codons in a highly non-random fashion. These codon usage biases sometimes frustrate attempts to express high levels of exogenous genes in hosts of widely divergent species. The Caenorhabditis elegans GluCl\u03b11 and GluCl\u03b2 genes form a functional glutamate and ivermectin-gated chloride channel when expressed in Xenopus oocytes, but expression is weak in mammalian cells. We have constructed synthetic genes that retain the amino acid sequence of the wild-type GluCl channel proteins, but use codons that are optimal for mammalian cell expression. We have tagged the native and codon-optimized GluCl cDNAs with enhanced yellow fluorescent protein (EYFP, GluCl\u03b11 subunit) and enhanced cyan fluorescent protein (EFCP, GluCl\u03b2 subunit), expressed the channels in E18 rat hippocampal neurons and measured the relative expression levels of the two genes with fluorescence microscopy as well as with electrophysiology. Codon optimization provides a 6- to 9-fold increase in expression, allowing the conclusions that the ivermectin-gated channel has an EC50 of 1.2 nM and a Hill coefficient of 1.9. We also confirm that the Y182F mutation in the codon-optimized \u03b2 subunit results in a heteromeric channel that retains the response to ivermectin while reducing the response to 100 \u03bcM glutamate by 7-fold. The engineered GluCl channel is the first codon-optimized membrane protein expressed in mammalian cells and may be useful for selectively silencing specific neuronal populations in vivo.", "date": "2003-03-30", "date_type": "published", "publication": "Journal of Neuroscience Methods", "volume": "124", "number": "1", "publisher": "Elsevier", "pagerange": "75-81", "id_number": "CaltechAUTHORS:20200626-123755093", "issn": "0165-0270", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200626-123755093", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "NIH", "grant_number": "MH 49176" }, { "agency": "Plum Foundation" }, { "agency": "William T. Gimbel Discovery Fund" } ] }, "doi": "10.1016/s0165-0270(02)00362-x", "resource_type": "article", "pub_year": "2003", "author_list": "Slimko, Eric M. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/b0w15-e5946", "eprint_id": 103861, "eprint_status": "archive", "datestamp": "2023-08-22 00:17:04", "lastmod": "2023-10-20 16:46:54", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Rozengurt-N", "name": { "family": "Rozengurt", "given": "Nora" } }, { "id": "Lopez-I", "name": { "family": "Lopez", "given": "Ivan" } }, { "id": "Chiu-Chi-Sung", "name": { "family": "Chiu", "given": "Chi-Sung" } }, { "id": "Kofuji-Paulo", "name": { "family": "Kofuji", "given": "Paulo" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Neusch-C", "name": { "family": "Neusch", "given": "Clemens" } } ] }, "title": "Time course of inner ear degeneration and deafness in mice lacking the Kir4.1 potassium channel subunit", "ispublished": "pub", "full_text_status": "public", "keywords": "Deafness; Inner ear; Stria vascularis; Inwardly rectifying K+ channel; Development; KCNJ10", "note": "\u00a9 2003 Elsevier Science B.V. \n\nReceived 6 September 2001, Accepted 20 December 2002, Available online 12 February 2003.", "abstract": "The Kir4.1 gene (KCNJ10) encodes an inwardly rectifying K\u207a channel subunit abundantly expressed in the CNS. Its expression in the mammalian inner ear has been suggested but its function in vivo in the inner ear is unknown. Because diverse human hereditary deafness syndromes are associated with mutations in K\u207a channels, we examined auditory function and inner ear structure in mice with a genetically inactivated Kir4.1 K\u207a channel subunit. Startle response experiments suggest that Kir4.1\u2212/\u2212 mice are profoundly deaf, whereas Kir4.1+/\u2212 mice react like wild-type mice to acoustic stimuli. In Kir4.1\u2212/\u2212 mice, the Reissner membrane is collapsed, the tectorial membrane is swollen, and type I hair cells and spiral ganglion neurons as well as their central processes degenerate over the first postnatal weeks. In the vestibular ganglia, neuronal cell death with apoptotic features is also observed. Immunostaining reveals that Kir4.1 is strongly expressed in stria vascularis of wild-type but not Kir4.1\u2212/\u2212 mice. Within the spiral ganglion, Kir4.1 labeling was detected on satellite cells surrounding spiral ganglion neurons and axons. We conclude that Kir4.1 is crucial for normal development of the cochlea and hearing, via two distinct aspects of extracellular K\u207a homeostasis: (1) in stria vascularis, Kir4.1 helps to generate the cochlear endolymph; and (2) in spiral and vestibular ganglia, Kir4.1 in surrounding glial cells helps to support the spiral and vestibular ganglion neurons and their projecting axons.", "date": "2003-03", "date_type": "published", "publication": "Hearing Research", "volume": "177", "number": "1-2", "publisher": "Elsevier", "pagerange": "71-80", "id_number": "CaltechAUTHORS:20200612-075804212", "issn": "0378-5955", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-075804212", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/s0378-5955(02)00799-2", "resource_type": "article", "pub_year": "2003", "author_list": "Rozengurt, Nora; Lopez, Ivan; et el." }, { "id": "https://authors.library.caltech.edu/records/wsqzd-1an69", "eprint_id": 27156, "eprint_status": "archive", "datestamp": "2023-08-19 10:54:14", "lastmod": "2023-10-24 16:55:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Leite-J-F", "name": { "family": "Leite", "given": "John F." } }, { "id": "Rodrigues-Pinguet-Nivalda-O", "name": { "family": "Rodrigues-Pinguet", "given": "Nivalda" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Insights into channel function via channel dysfunction", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2003 American Society for Clinical Investigation.\nPublished February 15, 2003.\n\nPublished - LEIjci03.pdf
", "abstract": "The nicotinic synapse has been a touchstone for advances in neuroscience ever since Jean Nicot, the French ambassador to Portugal, sent some tobacco seeds home to Paris in 1550 with a note that the New World plant had interesting effects when smoked. Now the muscle nicotinic acetylcholine receptor (nAChR) is a well-studied example of ligand-gated ion channels. After a motor neuron is stimulated, the nerve impulse reaches the presynaptic terminal, where it evokes release of acetylcholine (ACh) into the synapse. The nAChR depolarizes the postsynaptic muscle and triggers muscle action potentials; muscle contraction follows. To date, several nAChR subtypes have been successfully isolated, purified, imaged, and expressed, and unitary currents have been recorded from these channels (1). Researchers continue to unravel the molecular mechanisms of these macromolecules that are embedded in membranes at vertebrate nerve-muscle synapses, at invertebrate nicotinic synapses (which explains why nicotine-producing tobacco plants have a select advantage against invertebrate pests), and in the vertebrate central system (which explains Jean Nicot's fascination with those leaves). However, the precise structural events that trigger channel opening or \"gating\" remain mostly unknown.", "date": "2003-02", "date_type": "published", "publication": "Journal of Clinical Investigation", "volume": "111", "number": "4", "publisher": "American Society for Clinical Investigation", "pagerange": "436-437", "id_number": "CaltechAUTHORS:20111011-100601324", "issn": "0021-9738", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20111011-100601324", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1172/JCI200317882", "pmcid": "PMC151932", "primary_object": { "basename": "LEIjci03.pdf", "url": "https://authors.library.caltech.edu/records/wsqzd-1an69/files/LEIjci03.pdf" }, "resource_type": "article", "pub_year": "2003", "author_list": "Leite, John F.; Rodrigues-Pinguet, Nivalda; et el." }, { "id": "https://authors.library.caltech.edu/records/m3gnb-tp657", "eprint_id": 98955, "eprint_status": "archive", "datestamp": "2023-08-21 23:56:51", "lastmod": "2023-10-18 17:45:29", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Chiu-Chi-Sung", "name": { "family": "Chiu", "given": "Chi-Sung" } }, { "id": "Jensen-Kimmo", "name": { "family": "Jensen", "given": "Kimmo" } }, { "id": "Sokolova-Irina", "name": { "family": "Sokolova", "given": "Irina" } }, { "id": "Wang-Daniel", "name": { "family": "Wang", "given": "Dan" } }, { "id": "Li-Ming", "name": { "family": "Li", "given": "Ming" } }, { "id": "Deshpande-Purnima", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Mody-Istvan", "name": { "family": "Mody", "given": "Istvan" } }, { "id": "Quick-Michael-W", "name": { "family": "Quick", "given": "Michael W." } }, { "id": "Quake-S-R", "name": { "family": "Quake", "given": "Stephen R." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Number, Density, and Surface/Cytoplasmic Distribution of GABA Transporters at Presynaptic Structures of Knock-In Mice Carrying GABA Transporter Subtype 1\u2013Green Fluorescent Protein Fusions", "ispublished": "pub", "full_text_status": "public", "keywords": "GABA; synapse; transporter; green fluorescent protein; mouse; knock-in", "note": "\u00a9 2002 Society for Neuroscience. \n\nReceived June 4, 2002; revised Aug. 8, 2002; accepted Sept. 11, 2002. \n\nThis research was supported by grants from National Institutes of Health (DA-09121, DA-010509, NS-11756, MH-49176, MH-61468, NS-030549, and National Research Service Award to M.L.) and the National Science Foundation (0119493), and by a Della Martin Fellowship (C.-S.C.). We are indebted to Cesar Labarca and other members of the Caltech and University of California Los Angeles groups for much advice, Nathan Nelson for the mGAT1 cDNA, Tau-Mu Yi and M. Simon for use and help with the confocal microscope, and Melinda Turner and Robert Farley for discussion.\n\nPublished - 10251.full.pdf
", "abstract": "GABA transporter subtype 1 (GAT1) molecules were counted near GABAergic synapses, to a resolution of \u223c0.5 \u03bcm. Fusions between GAT1 and green fluorescent protein (GFP) were tested in heterologous expression systems, and a construct was selected that shows function, expression level, and trafficking similar to that of wild-type (WT) GAT1. A strain of knock-in mice was constructed that expresses this mGAT1\u2013GFP fusion in place of the WT GAT1 gene. The pattern of fluorescence in brain slices agreed with previous immunocytochemical observations. [^3H]GABA uptake, synaptic electrophysiology, and subcellular localization of the mGAT1\u2013GFP construct were also compared with WT mice. Quantitative fluorescence microscopy was used to measure the density of mGAT1\u2013GFP at presynaptic structures in CNS preparations from the knock-in mice. Fluorescence measurements were calibrated with transparent beads and gels that have known GFP densities. Surface biotinylation defined the fraction of transporters on the surface versus those in the nearby cytoplasm. The data show that the presynaptic boutons of GABAergic interneurons in cerebellum and hippocampus have a membrane density of 800\u20131300 GAT1 molecules per square micrometer, and the axons that connect boutons have a linear density of 640 GAT1 molecules per micrometer. A cerebellar basket cell bouton, a pinceau surrounding a Purkinje cell axon, and a cortical chandelier cell cartridge carry 9000, 7.8 million, and 430,000 GAT1 molecules, respectively; 61\u201363% of these molecules are on the surface membrane. In cultures from hippocampus, the set of fluorescent cells equals the set of GABAergic interneurons. Knock-in mice carrying GFP fusions of membrane proteins provide quantitative data required for understanding the details of synaptic transmission in living neurons.", "date": "2002-12-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "22", "number": "23", "publisher": "Society for Neuroscience", "pagerange": "10251-10266", "id_number": "CaltechAUTHORS:20190930-133756603", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190930-133756603", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "DA-010509" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "MH-61468" }, { "agency": "NIH", "grant_number": "NS-030549" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "NSF", "grant_number": "CBET-0119493" }, { "agency": "Della Martin Foundation" } ] }, "doi": "10.1523/jneurosci.22-23-10251.2002", "pmcid": "PMC6758747", "primary_object": { "basename": "10251.full.pdf", "url": "https://authors.library.caltech.edu/records/m3gnb-tp657/files/10251.full.pdf" }, "resource_type": "article", "pub_year": "2002", "author_list": "Chiu, Chi-Sung; Jensen, Kimmo; et el." }, { "id": "https://authors.library.caltech.edu/records/qpjkt-bb406", "eprint_id": 76579, "eprint_status": "archive", "datestamp": "2023-08-19 10:12:28", "lastmod": "2023-10-25 16:08:06", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Petersson-E-J", "name": { "family": "Petersson", "given": "E. James" }, "orcid": "0000-0003-3854-9210" }, { "id": "Choi-Anita", "name": { "family": "Choi", "given": "Anita" } }, { "id": "Dahan-D-S", "name": { "family": "Dahan", "given": "David S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "A Perturbed pK_a at the Binding Site of the Nicotinic Acetylcholine Receptor: Implications for Nicotine Binding", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2002 American Chemical Society. \n\nReceived 19 August 2002. Published online 5 October 2002. Published in print 1 October 2002. \n\nWe thank Dr. Lintong Li for her help. This work was supported by the National Institutes of Health (NS-34407 and NS-11756).\n\nSupplemental Material - ja028206i-2_s1.pdf
Supplemental Material - ja028206i_s1.pdf
", "abstract": "A series of tethered quaternary ammonium derivatives of Tyr have been incorporated into the binding site of the nicotinic acetylcholine receptor (nAChR) using the in vivo nonsense suppression method, producing constitutively active (self-gating) receptors. We have incorporated primary, secondary, and tertiary amine tethered agonists to give receptors whose constitutive activity can be modulated by pH. Lowering the pH protonates the tethered amine, giving it a positive charge and allowing it to reversibly activate the receptor. Tertiary and secondary tethered amines, TyrO3T and TyrO3S, have been successfully incorporated at \u03b1149 in the nAChR. Constitutive currents at pH 5.5 are 6 times those at pH 9.0. The pK_a of TyrO3T in the binding site appears to be 6 or lower, differing substantially from its pK_a in solution (\u223c9.3). This local pK_a perturbation has substantial implications for pharmacological research on the nAChR:\u2009 of the tertiary agonists considered, noracetylcholine experiences this pK_a perturbation, while nicotine does not.", "date": "2002-10-30", "date_type": "published", "publication": "Journal of the American Chemical Society", "volume": "124", "number": "43", "publisher": "American Chemical Society", "pagerange": "12662-12663", "id_number": "CaltechAUTHORS:20170414-144546207", "issn": "0002-7863", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170414-144546207", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1021/ja028206i", "primary_object": { "basename": "ja028206i-2_s1.pdf", "url": "https://authors.library.caltech.edu/records/qpjkt-bb406/files/ja028206i-2_s1.pdf" }, "related_objects": [ { "basename": "ja028206i_s1.pdf", "url": "https://authors.library.caltech.edu/records/qpjkt-bb406/files/ja028206i_s1.pdf" } ], "resource_type": "article", "pub_year": "2002", "author_list": "Petersson, E. James; Choi, Anita; et el." }, { "id": "https://authors.library.caltech.edu/records/ewk62-jre12", "eprint_id": 103868, "eprint_status": "archive", "datestamp": "2023-08-21 23:42:53", "lastmod": "2023-10-20 16:47:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Ping", "name": { "family": "Li", "given": "Ping" } }, { "id": "Slimko-E-M", "name": { "family": "Slimko", "given": "Eric M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Selective elimination of glutamate activation and introduction of fluorescent proteins into a Caenorhabditis elegans chloride channel", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Ion channel; Electrical silencing", "note": "\u00a9 2002 by the Biophysical Society. \n\nSubmitted September 24, 2001, and accepted for publication February 13, 2002. \n\nWe thank Kira Kostenko and Carrie Shilyansky for help with the oocytes and members of our research group for helpful discussion. This research was supported by a grant from the National Institutes of Health (NS\u201011756) and by the William T. Gimbel Discovery Fund in Neuroscience.", "abstract": "Glutamate\u2010gated chloride (GluCl) channels from invertebrates can be activated by ivermectin (IVM) to produce electrical silencing in mammalian neurons. To improve this GluCl/IVM strategy, we sought to mutate the Caenorhabditis elegans GluCl channels so that they become insensitive to glutamate but retain their sensitivity to IVM. Based on structure\u2013function studies of nicotinic acetylcholine receptor superfamily members, we tested in oocytes 19 point mutants at 16 residues in the \u03b2\u2010subunit likely to be involved in the response to glutamate. Y182F reduces the glutamate response by greater than six\u2010fold, with little change to IVM responses, when coexpressed with wild\u2010type (WT) GluCl \u03b1. For GluCl \u03b1\u03b2(Y182F), the EC\u2085\u2080 and Hill coefficient for glutamate are similar to those of WT, indicating that the mutant decreases the efficacy of glutamate, but not the potency. Also, fluorescent proteins (enhanced green fluorescent protein, enhanced yellow fluorescent protein, enhanced cyan fluorescent protein; XFP) were inserted into the M3\u2013M4 loop of the GluCl \u03b1, \u03b2 and \u03b2(Y182F). We found no significant functional difference between these XFP\u2010tagged receptors and WT receptors. The modified GluCl channel, without glutamate sensitivity but with a fluorescent tag, may be more useful in GluCl silencing strategies.", "date": "2002-09-25", "date_type": "published", "publication": "FEBS Letters", "volume": "528", "number": "1-3", "publisher": "Elsevier", "pagerange": "77-82", "id_number": "CaltechAUTHORS:20200612-113333592", "issn": "0014-5793", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-113333592", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS\u201011756" }, { "agency": "William T. Gimbel Discovery Fund" } ] }, "doi": "10.1016/s0014-5793(02)03245-3", "resource_type": "article", "pub_year": "2002", "author_list": "Li, Ping; Slimko, Eric M.; et el." }, { "id": "https://authors.library.caltech.edu/records/q45bq-nt875", "eprint_id": 55477, "eprint_status": "archive", "datestamp": "2023-08-21 23:41:06", "lastmod": "2023-10-20 22:17:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Slimko-Eric-M", "name": { "family": "Slimko", "given": "Eric M." } }, { "id": "McKinney-Sheri-L", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Anderson-D-J", "name": { "family": "Anderson", "given": "David J." }, "orcid": "0000-0001-6175-3872" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Selective Electrical Silencing of Mammalian Neurons In Vitro by the Use of Invertebrate Ligand-Gated Chloride Channels", "ispublished": "pub", "full_text_status": "public", "keywords": "silencing; excitability; hippocampal neurons; chloride channel; Sindbis virus; transfection of neurons; EGFP; EYFP; ECFP", "note": "\u00a9 2002 Society for Neuroscience.\n\nReceived March 25, 2002; Revised June 5, 2002; Accepted June 13, 2002.\n\nThis work was supported by National Institutes of Health Grants NS 11756 and MH 49176, by the Sidney Stern and Plum Foundations, and by the William T. Gimbal Discovery fund in Neuroscience. We thank Doris Cully for generously supplying the C. elegans GluCl channel cDNAs and for discussion; Birgit Hirschberg, Charles Cohen, and Christof Koch for discussion; and Dong Ju for technical assistance.\n\nPublished - 7373.full.pdf
", "abstract": "Selectively reducing the excitability of specific neurons will (1) allow for the creation of animal models of human neurological disorders and (2) provide insight into the global function of specific sets of neurons. We focus on a combined genetic and pharmacological approach to silence neurons electrically. We express invertebrate ivermectin (IVM)-sensitive chloride channels (Caenorhabditis elegans GluCl \u03b1 and \u03b2) with a Sindbis virus and then activate these channels with IVM to produce inhibition via a Cl^\u2212 conductance. We constructed a three-cistron Sindbis virus that expresses the \u03b1 and \u03b2 subunits of a glutamate-gated chloride channel (GluCl) along with the green fluorescent protein (EGFP) marker. Expression of the C. elegans channel does not affect the normal spike activity or GABA/glutamate postsynaptic currents of cultured embryonic day 18 hippocampal neurons. At concentrations as low as 5 nM, IVM activates a Cl^\u2212 current large enough to silence infected neurons effectively. This conductance reverses in 8 hr. These low concentrations of IVM do not potentiate GABA responses. Comparable results are observed with plasmid transfection of yellow fluorescent protein-tagged (EYFP) GluCl \u03b1 and cyan fluorescent protein-tagged (ECFP) GluCl \u03b2. The present study provides an in vitromodel mimicking conditions that can be obtained in transgenic mice and in viral-mediated gene therapy. These experiments demonstrate the feasibility of using invertebrate ligand-activated Cl^\u2212 channels as an approach to modulate excitability.", "date": "2002-09-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "22", "number": "17", "publisher": "Society for Neuroscience", "pagerange": "7373-7379", "id_number": "CaltechAUTHORS:20150303-122051974", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150303-122051974", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 11756" }, { "agency": "NIH", "grant_number": "MH 49176" }, { "agency": "Sidney Stern Memorial Trust" }, { "agency": "Plum Foundation" }, { "agency": "William T. Gimbel Discovery Fund" } ] }, "doi": "10.1523/JNEUROSCI.22-17-07373.2002", "pmcid": "PMC6757961", "primary_object": { "basename": "7373.full.pdf", "url": "https://authors.library.caltech.edu/records/q45bq-nt875/files/7373.full.pdf" }, "resource_type": "article", "pub_year": "2002", "author_list": "Slimko, Eric M.; McKinney, Sheri; et el." }, { "id": "https://authors.library.caltech.edu/records/qhh4f-vqp59", "eprint_id": 73717, "eprint_status": "archive", "datestamp": "2023-08-19 09:53:27", "lastmod": "2023-10-24 16:22:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Beene-D-L", "name": { "family": "Beene", "given": "Darren L." } }, { "id": "Brandt-G-S", "name": { "family": "Brandt", "given": "Gabriel S." } }, { "id": "Zhong-Wenge", "name": { "family": "Zhong", "given": "Wenge" } }, { "id": "Zacharias-N-M", "name": { "family": "Zacharias", "given": "Niki M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Cation\u2212\u03c0 Interactions in Ligand Recognition by Serotonergic (5-HT_(3A)) and Nicotinic Acetylcholine Receptors: The Anomalous Binding Properties of Nicotine", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2002 American Chemical Society. \n\nReceived 8 April 2002. Published online 19 July 2002. Published in print 1 August 2002.", "abstract": "A series of tryptophan analogues has been introduced into the binding site regions of two ion channels, the ligand-gated nicotinic acetylcholine and serotonin 5-HT_(3A) receptors, using unnatural amino acid mutagenesis and heterologous expression in Xenopus oocytes. A cation\u2212\u03c0 interaction between serotonin and Trp183 of the serotonin channel 5-HT3AR is identified for the first time, precisely locating the ligand-binding site of this receptor. The energetic contribution of the observed cation\u2212\u03c0 interaction between a tryptophan and the primary ammonium ion of serotonin is estimated to be approximately 4 kcal/mol, while the comparable interaction with the quaternary ammonium of acetylcholine is approximately 2 kcal/mol. The binding mode of nicotine to the nicotinic receptor of mouse muscle is examined by the same technique and found to differ significantly from that of the natural agonist, acetylcholine.", "date": "2002-08-01", "date_type": "published", "publication": "Biochemistry", "volume": "41", "number": "32", "publisher": "American Chemical Society", "pagerange": "10262-10269", "id_number": "CaltechAUTHORS:20170125-110549324", "issn": "0006-2960", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170125-110549324", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1021/bi020266d", "resource_type": "article", "pub_year": "2002", "author_list": "Beene, Darren L.; Brandt, Gabriel S.; et el." }, { "id": "https://authors.library.caltech.edu/records/8sq9k-pjz03", "eprint_id": 6242, "eprint_status": "archive", "datestamp": "2023-08-21 23:34:15", "lastmod": "2023-10-16 20:12:34", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Eickhorst-A-N", "name": { "family": "Eickhorst", "given": "Angela N." } }, { "id": "Berson-Amy", "name": { "family": "Berson", "given": "Amy" } }, { "id": "Cockayne-D", "name": { "family": "Cockayne", "given": "Debra" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Khahk-B-S", "name": { "family": "Khahk", "given": "Baljit S." } } ] }, "title": "Control of P2X2 Channel Permeability by the Cytosolic Domain", "ispublished": "pub", "full_text_status": "public", "keywords": "ATP; ion channel; modulation; P2X; purinoceptor", "note": "\u00a9 2002 Rockefeller University Press. fter the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. \n\nSubmitted: 20 November 2001. Revised: 13 May 2002. Accepted: 15 May 2002. Published online 15 July 2002 doi:10.1085/jgp.20028535 \n\nWe thank K. Kostenko and H. Li for help with preparation of Xenopus oocytes, G. Shapovalov for measurements of 2-(methyl-amino)-ethanol mobility, other members of the group for comments, and Nigel Unwin for discussion and comments on the paper. \n\nWork in our labs was supported by a Wellcome Trust (UK) Prize Fellowship, Roche Bioscience, National Institutes of Health (NS-11756), and the Medical Research Council.\n\nPublished - EICjgp02.pdf
", "abstract": "ATP-gated P2X channels are the simplest of the three families of transmitter-gated ion channels. Some P2X channels display a time- and activation-dependent change in permeability as they undergo the transition from the relatively Na+-selective I1 state to the I2 state, which is also permeable to organic cations. We report that the previously reported permeability change of rat P2X2 (rP2X2) channels does not occur at mouse P2X2 (mP2X2) channels expressed in oocytes. Domain swaps, species chimeras, and point mutations were employed to determine that two specific amino acid residues in the cytosolic tail domain govern this difference in behavior between the two orthologous channels. The change in pore diameter was characterized using reversal potential measurements and excluded field theory for several organic ions; both rP2X2 and mP2X2 channels have a pore diameter of ~11 \u00c5 in the I1 state, but the transition to the I2 state increases the rP2X2 diameter by at least 3 \u00c5. The I1 to I2 transition occurs with a rate constant of ~0.5 s^-1. The data focus attention on specific residues of P2X2 channel cytoplasmic domains as determinants of permeation in a state-specific manner.", "date": "2002-08", "date_type": "published", "publication": "Journal of General Physiology", "volume": "120", "number": "2", "publisher": "Rockefeller University Press", "pagerange": "119-131", "id_number": "CaltechAUTHORS:EICjgp02", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:EICjgp02", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1085/jgp.20028535", "pmcid": "PMC2234464", "primary_object": { "basename": "EICjgp02.pdf", "url": "https://authors.library.caltech.edu/records/8sq9k-pjz03/files/EICjgp02.pdf" }, "resource_type": "article", "pub_year": "2002", "author_list": "Eickhorst, Angela N.; Berson, Amy; et el." }, { "id": "https://authors.library.caltech.edu/records/z19tj-kww95", "eprint_id": 103870, "eprint_status": "archive", "datestamp": "2023-08-19 09:47:03", "lastmod": "2023-10-20 16:47:27", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Ming", "name": { "family": "Li", "given": "Ming" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Early Fluorescence Signals Detect Transitions at Mammalian Serotonin Transporters", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2002 The Biophysical Society. \n\nSubmitted September 24, 2001, and accepted for publication February 13, 2002. \n\nWe thank Yanhe Tong for making the C109A mutant of hSERT. \n\nThis research was supported by National Institutes of Health Grant DA-019121 and by a National Research Service Award to Ming Li.", "abstract": "The mammalian serotonin transporters rSERT or hSERT were expressed in oocytes and labeled with sulforhodamine-MTS. The endogenous Cys-109 residue contributes most of the signal, and the labeled transporter shows normal function. The SERT fluorescence decreases in the presence of 5-HT and also depends on the inorganic substrates of SERT. The fluorescence also increases with membrane depolarization. During voltage-jump experiments, fluorescence relaxations show little inactivation or history dependence. The fluorescence signal has a voltage dependence similar to that of the prepriming step of the previously described voltage-dependent transient current. However, the fluorescence relaxations are the fastest voltage-dependent events yet studied at SERT; their time constants of \u223c8\u201330 ms are severalfold faster than the prepriming or inactivation phases of the transient currents. These fluorescence signals are interpreted within the framework of the gate-lumen-gate model. The signals may monitor initial events at the outer gate.", "date": "2002-07", "date_type": "published", "publication": "Biophysical Journal", "volume": "83", "number": "1", "publisher": "Biophysical Society", "pagerange": "206-218", "id_number": "CaltechAUTHORS:20200612-113333889", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-113333889", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-019121" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1016/s0006-3495(02)75162-x", "pmcid": "PMC1302140", "resource_type": "article", "pub_year": "2002", "author_list": "Li, Ming and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/5rayc-kd131", "eprint_id": 103869, "eprint_status": "archive", "datestamp": "2023-08-19 09:46:56", "lastmod": "2023-10-20 16:47:17", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nadeau-H", "name": { "family": "Nadeau", "given": "H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "NRSF Causes cAMP-Sensitive Suppression of Sodium Current in Cultured Hippocampal Neurons", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2002 The American Physiological Society. \n\nReceived 20 December 2001; accepted in final form 7 March 2002. \n\nWe thank D. J. Anderson for useful suggestions and discussions and A. J. Paquette for critically reading the manuscript. \n\nThis work was supported by Burroughs-Wellcome and National Institute of Mental Health Grant MH-490176.\n\nThe costs of publication of this article were defrayed in part by the payment\nof page charges. The article must therefore be hereby marked \"advertisement\"\nin accordance with 18 U.S.C. Section 1734 solely to indicate this fact.", "abstract": "The neuron restrictive silencer factor (NRSF/REST) has been shown to bind to the promoters of many neuron-specific genes and is able to suppress transcription of Na\u207a channels in PC12 cells, although its functional effect in terminally differentiated neurons is unknown. We constructed lentiviral vectors to express NRSF as a bicistronic message with green fluorescent protein (GFP) and followed infected hippocampal neurons in culture over a period of 1\u20132 wk. NRSF-expressing neurons showed a time-dependent suppression of Na\u207a channel function as measured by whole cell electrophysiology. Suppression was reversed or prevented by the addition of membrane-permeable cAMP analogues and enhanced by cAMP antagonists but not affected by increasing protein expression with a viral enhancer. Secondary effects, including altered sensitivity to glutamate and GABA and reduced outward K\u207a currents, were duplicated by culturing GFP-infected control neurons in TTX. The striking similarity of the phenotypes makes NRSF potentially useful as a genetic \"silencer\" and also suggests avenues of further exploration that may elucidate the transcription factor's in vivo role in neuronal plasticity.", "date": "2002-07", "date_type": "published", "publication": "Journal of Neurophysiology", "volume": "88", "number": "1", "publisher": "American Physiological Society", "pagerange": "409-421", "id_number": "CaltechAUTHORS:20200612-113333802", "issn": "0022-3077", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-113333802", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Burroughs Wellcome Fund" }, { "agency": "NIH", "grant_number": "MH-490176" } ] }, "doi": "10.1152/jn.2002.88.1.409", "resource_type": "article", "pub_year": "2002", "author_list": "Nadeau, H. and Lester, H. A." }, { "id": "https://authors.library.caltech.edu/records/tc0w0-jaj87", "eprint_id": 7718, "eprint_status": "archive", "datestamp": "2023-08-21 23:10:30", "lastmod": "2023-10-23 15:58:47", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Petersson-E-J", "name": { "family": "Petersson", "given": "E. J." }, "orcid": "0000-0003-3854-9210" }, { "id": "Shahgholi-M", "name": { "family": "Shahgholi", "given": "M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "D. A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "MALDI-TOF mass spectrometry methods for evaluation of in vitro aminoacyl tRNA production", "ispublished": "pub", "full_text_status": "public", "keywords": "aminoacylation; gel electrophoresis; matrix-assisted laser desorption/ionization; nonsense suppression; T4 RNA ligase; T7 RNA polymerase; THG73 tRNA; unnatural amino acid", "note": "\u00a9 2002 RNA Society. \n\nReceived January 25, 2002; returned for revision February 1, 2002; revised manuscript received February 7, 2002. \n\nThe authors thank Dr. John F. Leite for his valuable input. This work was supported by the National Institutes of Health (NS-34407 and NS-11756).\n\nPublished - PETrna02.pdf
", "abstract": "Unnatural amino acid mutagenesis requires the in vitro production of aminoacyl tRNAs. Bacteriophage T4 RNA ligase is used to ligate a-amino-protected dCA amino acids to 74mer tRNA. Previously, there has been no facile method for evaluating the efficiency of this reaction prior to using the tRNA in translation. We report a novel use of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry in monitoring the formation of aminoacyl 76mer tRNA. This method is more efficient and precise than the traditional technique of gel electrophoresis. These MALDI conditions should also prove useful for analyzing aminoacyl tRNAs produced through aminoacyl tRNA synthetases and other methods.", "date": "2002-04", "date_type": "published", "publication": "RNA", "volume": "8", "number": "4", "publisher": "RNA", "pagerange": "542-547", "id_number": "CaltechAUTHORS:PETrnao02", "issn": "1355-8382", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:PETrnao02", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "pmcid": "PMC1370275", "primary_object": { "basename": "PETrna02.pdf", "url": "https://authors.library.caltech.edu/records/tc0w0-jaj87/files/PETrna02.pdf" }, "resource_type": "article", "pub_year": "2002", "author_list": "Petersson, E. J.; Shahgholi, M.; et el." }, { "id": "https://authors.library.caltech.edu/records/pja78-rpq32", "eprint_id": 103871, "eprint_status": "archive", "datestamp": "2023-08-21 23:04:38", "lastmod": "2023-10-20 16:47:34", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Ming", "name": { "family": "Li", "given": "Ming" } }, { "id": "Farley-R-A", "name": { "family": "Farley", "given": "Robert A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Voltage-dependent transient currents of human and rat 5-HT transporters (SERT) are blocked by HEPES and ion channel ligands", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Ion channel; Voltage jump", "note": "\u00a9 2015 Federation of European Biochemical Societies. \n\nReceived 17 December 2001; revised 17 January 2002; accepted 17 January 2002. First published online 31 January 2002. \n\nWe thank Chi\u2010Sung Chiu and Ken Philipson for helpful discussion. This work was supported by a grant from the National Institutes of Health (DA\u201009121) and by an NRSA to M.L.", "abstract": "The hyperpolarization\u2010activated transient current of mammalian 5\u2010hydroxytryptamine transporters (SERT) expressed in Xenopus oocytes was studied. Human (h) and rat (r) SERT transient currents are blocked by HEPES with changes in the waveform kinetics, and the blockade of hSERT has use\u2010dependent properties. HEPES also changes the time course of the prepriming step, especially for hSERT. Transient currents at hSERT and rSERT are also blocked by spermine and spermidine in the mM range, and by fluoxetine, cocaine, QX\u2010314, and QX\u2010222 in the \u03bcM range. These pharmacological and kinetic properties of transient current blockade emphasize the similarities between the transient current and phenomena at ion channels.", "date": "2002-02-27", "date_type": "published", "publication": "FEBS Letters", "volume": "513", "number": "2-3", "publisher": "Elsevier", "pagerange": "247-252", "id_number": "CaltechAUTHORS:20200612-113333997", "issn": "0014-5793", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-113333997", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA\u201009121" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM08042" } ] }, "doi": "10.1016/s0014-5793(02)02322-0", "resource_type": "article", "pub_year": "2002", "author_list": "Li, Ming; Farley, Robert A.; et el." }, { "id": "https://authors.library.caltech.edu/records/8re1n-adt23", "eprint_id": 103872, "eprint_status": "archive", "datestamp": "2023-08-19 08:57:15", "lastmod": "2023-10-20 16:47:40", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kovoor-A", "name": { "family": "Kovoor", "given": "Abraham" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Gi Irks GIRKs", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2002 Cell Press. Under an Elsevier user license. \n\nAvailable online 5 January 2002.", "abstract": "G protein-activated potassium channels (GIRKs), monitored with the temporal and molecular resolution of electrophysiology, play a key role in the study of signal transduction. GIRKs are activated primarily by the G\u03b2\u03b3 subunits, but a paper by Peleg et al. (2002)([this issue of Neuron]) demonstrates a role for G\u03b1 subunits in suppressing basal activity and supports the idea of a macromolecular complex of G protein, GIRK, and perhaps RGS protein.", "date": "2002-01-03", "date_type": "published", "publication": "Neuron", "volume": "33", "number": "1", "publisher": "Cell Press", "pagerange": "6-8", "id_number": "CaltechAUTHORS:20200612-113334102", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-113334102", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/s0896-6273(01)00572-4", "resource_type": "article", "pub_year": "2002", "author_list": "Kovoor, Abraham and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/hw8dz-dwb77", "eprint_id": 64387, "eprint_status": "archive", "datestamp": "2023-08-21 22:29:04", "lastmod": "2023-10-17 21:16:04", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Neusch-C", "name": { "family": "Neusch", "given": "Clemens" } }, { "id": "Rozengurt-N", "name": { "family": "Rozengurt", "given": "Nora" } }, { "id": "Jacobs-R-E", "name": { "family": "Jacobs", "given": "Russell E." }, "orcid": "0000-0002-1382-8486" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Kofuji-P", "name": { "family": "Kofuji", "given": "Paulo" } } ] }, "title": "Kir4.1 Potassium Channel Subunit Is Crucial for Oligodendrocyte Development and In Vivo Myelination", "ispublished": "pub", "full_text_status": "public", "keywords": "oligodendrocytes; myelination; inwardly rectifying potassium channels; knock-out mouse; glia; spinal cord", "note": "\u00a9 2001 Society for Neuroscience. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). \n\nReceived Jan. 12, 2001; revised May 8, 2001; accepted May 9, 2001.\n\nThis work was supported by National Institutes of Health Grants GM-29836, EY12949, DA08944, and RR13625 and the Deutsche Forschungsgemeinschaft (NE-767/1-1). We thank S. S. Velan for help with the magnetic resonance imaging, S. McKinney for help with animals, V. Santoro for help in data analysis, and B. S. Khakh for comments.\n\nPublished - 5429.full.pdf
", "abstract": "To understand the cellular and in vivo functions of specific K^+ channels in glia, we have studied mice with a null mutation in the weakly inwardly rectifying K^+ channel subunit Kir4.1. Kir4.1\u2212/\u2212 mice display marked motor impairment, and the cellular basis is hypomyelination in the spinal cord, accompanied by severe spongiform vacuolation, axonal swellings, and degeneration. Immunostaining in the spinal cord of wild-type mice up to postnatal day 18 reveals that Kir4.1 is expressed in myelin-synthesizing oligodendrocytes, but probably not in neurons or glial fibrillary acidic protein-positive (GFAP-positive) astrocytes. Cultured oligodendrocytes from developing spinal cord of Kir4.1\u2212/\u2212 mice lack most of the wild-type K^+ conductance, have depolarized membrane potentials, and display immature morphology. By contrast, cultured neurons from spinal cord of Kir4.1\u2212/\u2212 mice have normal physiological characteristics. We conclude that Kir4.1 forms the major K^+ conductance of oligodendrocytes and is therefore crucial for myelination. The Kir4.1 knock-out mouse is one of the few CNS dysmyelinating or demyelinating phenotypes that does not involve a gene directly involved in the structure, synthesis, degradation, or immune response to myelin. Therefore, this mouse shows how an ion channel mutation could contribute to the polygenic demyelinating diseases.", "date": "2001-08-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "21", "number": "15", "publisher": "Society for Neuroscience", "pagerange": "5429-5438", "id_number": "CaltechAUTHORS:20160210-145519981", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160210-145519981", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "EY12949" }, { "agency": "NIH", "grant_number": "DA08944" }, { "agency": "NIH", "grant_number": "RR13625" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)", "grant_number": "NE-767/1-1" } ] }, "doi": "10.1523/JNEUROSCI.21-15-05429.2001", "pmcid": "PMC6762664", "primary_object": { "basename": "5429.full.pdf", "url": "https://authors.library.caltech.edu/records/hw8dz-dwb77/files/5429.full.pdf" }, "resource_type": "article", "pub_year": "2001", "author_list": "Neusch, Clemens; Rozengurt, Nora; et el." }, { "id": "https://authors.library.caltech.edu/records/55r9j-brd89", "eprint_id": 103029, "eprint_status": "archive", "datestamp": "2023-08-21 22:23:15", "lastmod": "2023-10-20 00:42:55", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Philipson-K-D", "name": { "family": "Philipson", "given": "Kenneth D." } }, { "id": "Gallivan-J-P", "name": { "family": "Gallivan", "given": "Justin P." } }, { "id": "Brandt-G-S", "name": { "family": "Brandt", "given": "Gabriel S." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Incorporation of caged cysteine and caged tyrosine into a transmembrane segment of the nicotinic ACh receptor", "ispublished": "pub", "full_text_status": "restricted", "keywords": "nicotinic acetylcholine receptor; ion channel; site-directed mutagenesis; nonsense suppression; o-nitrobenzyl tyrosine; o-nitrobenzyl cysteine", "note": "\u00a9 2001 the American Physiological Society. \n\nReceived 26 October 2000; Accepted 28 February 2001; Published online 1 July 2001; Published in print 1 July 2001. \n\nWe thank Dr. B. Khakh for helpful discussions. \n\nThis research was supported by grants from the National Institutes of Health (NS-11756, NS-34407, and HL-49101). \n\nThe costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.", "abstract": "The nonsense codon suppression technique was used to incorporate o-nitrobenzyl cysteine or o-nitrobenzyl tyrosine (caged Cys or Tyr) into the 9\u2032 position of the M2 transmembrane segment of the \u03b3-subunit of the muscle nicotinic ACh receptor expressed in Xenopusoocytes. The caged amino acids replaced an endogenous Leu residue that has been implicated in channel gating. ACh-induced current increased substantially after ultraviolet (UV) irradiation to remove the caging group. This represents the first successful incorporation of caged Cys into a protein in vivo and the first incorporation of caged amino acids within a transmembrane segment of a membrane protein. The bulky nitrobenzyl group does not prevent the synthesis, assembly, or trafficking of the ACh receptor. When side chains were decaged using 1-ms UV light flashes, the channels with caged Cys or caged Tyr responded with strikingly different kinetics. The increase in current upon photolysis of caged Cys was too rapid for resolution by the voltage-clamp circuit [time constant (\u03c4) <10 ms], whereas the increase in current upon photolysis of caged Tyr was dominated by a phase with \u03c4 \u223c500 ms. Apparently, the presence of a bulkyo-nitrobenzyl Tyr residue distorts the receptor into an abnormal conformation. Upon release of the caging group, the receptor relaxes, with \u03c4 \u223c500 ms, into a conformation that allows the channel to open. Tyr at the 9\u2032 position of the \u03b3-subunit greatly increases the ability of ACh to block the channel by binding within the channel pore. This is manifested in two ways. 1) A \"rebound,\" or increase in current, occurs upon removal of ACh from the bathing medium; and 2) at ACh concentrations >400 \u03bcM, inward currents are decreased through the mutated channel. The ability to incorporate caged amino acids into proteins should have widespread utility.", "date": "2001-07", "date_type": "published", "publication": "American Journal of Physiology. Cell Physiology", "volume": "281", "number": "1", "publisher": "American Physiological Society", "pagerange": "C195-C206", "id_number": "CaltechAUTHORS:20200506-103644435", "issn": "0363-6143", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200506-103644435", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "HL-49101" } ] }, "doi": "10.1152/ajpcell.2001.281.1.c195", "resource_type": "article", "pub_year": "2001", "author_list": "Philipson, Kenneth D.; Gallivan, Justin P.; et el." }, { "id": "https://authors.library.caltech.edu/records/wrav9-3x965", "eprint_id": 103873, "eprint_status": "archive", "datestamp": "2023-08-19 07:51:56", "lastmod": "2023-10-20 16:47:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Ming", "name": { "family": "Li", "given": "Ming" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Ion Channel Diseases of the Central Nervous System", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Ion channels \u2013 Epilepsy \u2013 Ataxia \u2013 Schizophrenia \u2014 Alzheimer's disease", "note": "\u00a9 2001 Neva Press. \n\nWe thank Baljit Khakh and Wenmei Shi for comments. Work in this laboratory is supported by grants from the NIH (DA09121, NS11756, GM29836, MH49176), the University of California Tobacco Related Disease Research Program, and the National Parkinson Foundation. Ming Li holds a National Research Service Award.", "abstract": "In the last decade, advances in molecular genetics and cellular electrophysiology have increased our understanding of ion channel function. A number of diseases termed \"channelopathies\" have been discovered that are caused by ion channel dysfunction. Channelopathies can be caused by autoimmune, iatrogenic, toxic or genetic mechanisms. Mutations in genes encoding ion channel proteins that disrupt channel function are now the most commonly identified cause of channelopathies, perhaps because gene disruption is readily detected by the methods of molecular genetics. Ion channels are abundant in the central nervous system (CNS), but CNS channelopathies are rare; however, they overlap with some important neurological disorders, such as epilepsy, ataxia, migraine, schizophrenia, Alzheimer's disease and other neurodegenerative diseases. It is possible that more CNS channelopathies will be discovered when additional ion channels are characterized and the complex mechanisms of brain function are better understood. At present, increased knowledge of the identity, structure and function of ion channels is facilitating diagnosis and treatment of many channelopathies.", "date": "2001-06", "date_type": "published", "publication": "CNS Drug Reviews", "volume": "7", "number": "2", "publisher": "Wiley", "pagerange": "214-240", "id_number": "CaltechAUTHORS:20200612-115646273", "issn": "1080-563X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-115646273", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA09121" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "MH49176" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "National Parkinson Foundation" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1111/j.1527-3458.2001.tb00196.x", "pmcid": "PMC6741647", "resource_type": "article", "pub_year": "2001", "author_list": "Li, Ming and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/e4v39-y2847", "eprint_id": 56176, "eprint_status": "archive", "datestamp": "2023-08-19 07:44:25", "lastmod": "2023-10-20 23:44:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Neurobiology: Snails, synapses and smokers", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2001 Macmillan Magazines Ltd.", "abstract": "The discovery of a protein that controls the transmission of nerve impulses in snails is significant in its own right. It also advances our understanding of the vertebrate neurotransmitter receptor that responds to nicotine.", "date": "2001-05-17", "date_type": "published", "publication": "Nature", "volume": "411", "number": "6835", "publisher": "Nature Publishing Group", "pagerange": "252-254", "id_number": "CaltechAUTHORS:20150327-103308333", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150327-103308333", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1038/35077192", "resource_type": "article", "pub_year": "2001", "author_list": "Dougherty, Dennis A. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/9cv3w-vb247", "eprint_id": 961, "eprint_status": "archive", "datestamp": "2023-08-21 22:10:21", "lastmod": "2023-10-13 22:03:28", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Khakh-B-S", "name": { "family": "Khakh", "given": "Baljit S." } }, { "id": "Smith-W-B", "name": { "family": "Smith", "given": "W. Bryan" } }, { "id": "Chiu-Chi-Sung", "name": { "family": "Chiu", "given": "Chi-Sung" } }, { "id": "Ju-Donghong", "name": { "family": "Ju", "given": "Donghong" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Activation-dependent changes in receptor distribution and dendritic morphology in hippocampal neurons expressing P2X(2)-green fluorescent protein receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "ion channel, ATP, filopodia, ionotropic glutamate receptors, single-channel properties, gated cation channels, ion channels, P2X receptor, redistribution, purinoceptors, kinetics, cells, desensitization", "note": "\u00a9 2001 by the National Academy of Sciences. \n\nContributed by Norman Davidson, February 21, 2001. \n\nWe thank H. Li for preparation of Xenopus oocytes and E. Schuman for advice and use of the confocal microscope. This work was supported by a Wellcome Trust (U.K.) Prize Fellowship (to B.S.K.), by a Roche Bioscience Award, and by the National Institutes of Health (NS-11756, MH 49176). \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nFor the article \"Activation-dependent changes in receptor distribution and dendritic morphology in hippocampal neurons expressing P2X2-green fluorescent protein receptors\" by Baljit S. Khakh, W. Bryan Smith, Chi-Sung Chiu, Donghong Ju, Norman Davidson, and Henry A. Lester, which appeared in number 9, April 24, 2001, of Proc. Natl. Acad. Sci. USA (98, 5288\u20135293; First Published April 10, 2001; 10.1073/pnas.081089198), due to a printer's error, Baljit Khakh's e-mail was incorrectly listed as bsk@mrc-lmb.com.ac.uk. It should be ku.ca.mac.bml-crm@ksb.\n\nPublished - KHApnas01a.pdf
Erratum - KHApnas01errata.pdf
", "abstract": "ATP-gated P2X(2) receptors are widely expressed in neurons, but the cellular effects of receptor activation are unclear. We engineered functional green fluorescent protein (GFP)-tagged P2X(2) receptors and expressed them in embryonic hippocampal neurons, and report an approach to determining functional and total receptor pool sizes in living cells. ATP application to dendrites caused receptor redistribution and the formation of varicose hot spots of higher P2X(2)-GFP receptor density. Redistribution in dendrites was accompanied by an activation-dependent enhancement of the ATP-evoked current. Substate-specific mutant T18A P2X(2)-GFP receptors showed no redistribution or activation-dependent enhancement of the ATP-evoked current. Thus fluorescent P2X(2)-GFP receptors function normally, can be quantified, and reveal the dynamics of P2X(2) receptor distribution on the seconds time scale.", "date": "2001-04-24", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "98", "number": "9", "publisher": "National Academy of Sciences", "pagerange": "5288-5293", "id_number": "CaltechAUTHORS:KHApnas01a", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:KHApnas01a", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "Roche Biosciences" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "MH 49176" } ] }, "doi": "10.1073/pnas.081089198", "pmcid": "PMC33202", "primary_object": { "basename": "KHApnas01a.pdf", "url": "https://authors.library.caltech.edu/records/9cv3w-vb247/files/KHApnas01a.pdf" }, "related_objects": [ { "basename": "KHApnas01errata.pdf", "url": "https://authors.library.caltech.edu/records/9cv3w-vb247/files/KHApnas01errata.pdf" } ], "resource_type": "article", "pub_year": "2001", "author_list": "Khakh, Baljit S.; Smith, W. Bryan; et el." }, { "id": "https://authors.library.caltech.edu/records/d2t2m-1zx50", "eprint_id": 895, "eprint_status": "archive", "datestamp": "2023-08-21 22:10:10", "lastmod": "2023-10-13 22:01:57", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Yu-Tzu-ping", "name": { "family": "Yu", "given": "Tzu-ping" } }, { "id": "McKinney-S", "name": { "family": "McKinney", "given": "Sheri" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "gamma-Aminobutyric acid type A receptors modulate cAMP-mediated long-term potentiation and long-term depression at monosynaptic CA3-CA1 synapses", "ispublished": "pub", "full_text_status": "public", "keywords": "SYNAPTIC ACTIVATION CAUSES, DEPENDENT PROTEIN-KINASE, HIPPOCAMPAL CA1 NEURONS, CYCLIC-AMP, LATE-PHASE, LTP, ADENOSINE, TRANSMISSION, STIMULATION, CULTURES", "note": "\u00a9 2001 by the National Academy of Sciences. \n\nContributed by Norman Davidson, February 23, 2001. \n\nWe thank Sami Barghshoon for preparing the slice cultures. This work was supported by the National Institutes of Health and the McKnight Foundation. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - PUTpnas01.pdf
", "abstract": "cAMP induces a protein-synthesis-dependent late phase of longterm potentiation (LTP) at CA3-CA1 synapses in acute hippocampal slices. Herein we report cAMP-mediated LTP and long-term depression (LTD) at monosynaptic CA3-CA1 cell pairs in organotypic hippocampal slice cultures. After bath application of the membrane-permeable cAMP analog adenosine 3 ' ,5 ' -cyclic monophosphorothioate, Sp isomer (Sp-cAMPS), synaptic transmission was enhanced for at least 2 h. Consistent with previous findings, the late phase of LTP requires activation of cAMP-dependent protein kinase A and protein synthesis. There is also an early phase of LTP induced by cAMP; the early phase depends on protein kinase A but, in contrast to the later phase, does not require protein synthesis, in addition, the cAMP-induced LTP is associated with a reduction of paired-pulse facilitation, suggesting that presynaptic modification may be involved. Furthermore, we found that Sp-cAMPS induced LTD in slices pretreated with picrotoxin, a gamma -aminobutyric acid type A (GABA(A)) receptor antagonist. This form of LTD depends on protein synthesis and protein phosphatase(s) and is accompanied by an increased ratio of failed synaptic transmission. These results suggest that GABA(A) receptors can modulate the effect of cAMP on synaptic transmission and thus determine the direction of synaptic plasticity.", "date": "2001-04-24", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "98", "number": "9", "publisher": "National Academy of Sciences", "pagerange": "5264-5269", "id_number": "CaltechAUTHORS:YUTpnas01", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:YUTpnas01", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "McKnight Foundation" } ] }, "doi": "10.1073/pnas.091093998", "pmcid": "PMC33198", "primary_object": { "basename": "PUTpnas01.pdf", "url": "https://authors.library.caltech.edu/records/d2t2m-1zx50/files/PUTpnas01.pdf" }, "resource_type": "article", "pub_year": "2001", "author_list": "Yu, Tzu-ping; McKinney, Sheri; et el." }, { "id": "https://authors.library.caltech.edu/records/880c8-dzp70", "eprint_id": 901, "eprint_status": "archive", "datestamp": "2023-08-21 22:01:37", "lastmod": "2023-10-13 22:02:08", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Schwarz-J", "name": { "family": "Schwarz", "given": "Johannes" } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Schwarz-S", "name": { "family": "Schwarz", "given": "Sigrid" } }, { "id": "Nowak-M-W", "name": { "family": "Nowak", "given": "Mark W." } }, { "id": "Fonck-C", "name": { "family": "Fonck", "given": "Carlos" } }, { "id": "Nashmi-R", "name": { "family": "Nashmi", "given": "Raad" } }, { "id": "Kofuji-Paulo", "name": { "family": "Kofuji", "given": "Paulo" } }, { "id": "Dang-Hong", "name": { "family": "Dang", "given": "Hong" } }, { "id": "Shi-Wenmei", "name": { "family": "Shi", "given": "Wenmei" } }, { "id": "Fidan-M", "name": { "family": "Fidan", "given": "Melihat" } }, { "id": "Khakh-B-S", "name": { "family": "Khakh", "given": "Baljit S." } }, { "id": "Chen-Zhoufeng", "name": { "family": "Chen", "given": "Zhoufeng" } }, { "id": "Bowers-B-J", "name": { "family": "Bowers", "given": "Barbara J." } }, { "id": "Boulter-J", "name": { "family": "Boulter", "given": "Jim" } }, { "id": "Wehner-J-M", "name": { "family": "Wehner", "given": "Jeanne M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Point mutant mice with hypersensitive alpha 4 nicotinic receptors show dopaminergic deficits and increased anxiety", "ispublished": "pub", "full_text_status": "public", "keywords": "RAT SUBSTANTIA-NIGRA, ACETYLCHOLINE-RECEPTORS, PARKINSONS-DISEASE, PROGENITOR CELLS, M2 DOMAIN, MODULATION, BEHAVIOR, LACKING, SUBUNIT, NEURONS", "note": "\u00a9 2001 by the National Academy of Sciences. \n\nCommunicated by Norman Davidson, California Institute of Technology, Pasadena, CA, December 8, 2000 (received for review October 4, 2000). \n\nWe thank Shirley Pease and her colleagues for generating and maintaining the mice. This research was supported by grants from the National Institutes of Health (NS-11756, MH-49176, DA-10156, and DA11836) and from the California Tobacco-Related Disease Research Program and by fellowships from the Alexander von Humboldt Foundation, the German Academic Exchange Foundation, and the Wellcome Trust. \n\nJ.S. and P.D. contributed equally to this work.\n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - LABpnas01.pdf
", "abstract": "Knock-in mice were generated that harbored a leucine-to-serine mutation in the alpha4 nicotinic receptor near the gate in the channel pore. Mice with intact expression of this hypersensitive receptor display dominant neonatal lethality. These mice have a severe deficit of dopaminergic neurons in the substantia nigra, possibly because the hypersensitive receptors are continuously activated by normal extracellular choline concentrations. A strain that retains the neo selection cassette in an intron has reduced expression of the hypersensitive receptor and is viable and fertile. The viable mice display increased anxiety, poor motor learning, excessive ambulation that is eliminated by very low levels of nicotine, and a reduction of nigrostriatal dopaminergic function upon aging. These knock-in mice provide useful insights into the pathophysiology of sustained nicotinic receptor activation and may provide a model for Parkinson's disease.", "date": "2001-02-27", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "98", "number": "5", "publisher": "National Academy of Sciences", "pagerange": "2786-2791", "id_number": "CaltechAUTHORS:LABpnas01", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LABpnas01", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "DA-10156" }, { "agency": "NIH", "grant_number": "DA-11836" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Alexander von Humboldt Foundation" }, { "agency": "Deutscher Akademischer Austauschdienst (DAAD)" }, { "agency": "Wellcome Trust" } ] }, "doi": "10.1073/pnas.041582598", "pmcid": "PMC30217", "primary_object": { "basename": "LABpnas01.pdf", "url": "https://authors.library.caltech.edu/records/880c8-dzp70/files/LABpnas01.pdf" }, "resource_type": "article", "pub_year": "2001", "author_list": "Labarca, Cesar; Schwarz, Johannes; et el." }, { "id": "https://authors.library.caltech.edu/records/rt275-vt526", "eprint_id": 5168, "eprint_status": "archive", "datestamp": "2023-08-21 21:58:34", "lastmod": "2023-10-16 19:04:13", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Tong-Yanhe", "name": { "family": "Tong", "given": "Yanhe" } }, { "id": "Brandt-G-S", "name": { "family": "Brandt", "given": "Gabriel S." } }, { "id": "Li-Ming", "name": { "family": "Li", "given": "Ming" } }, { "id": "Shapovalov-G", "name": { "family": "Shapovalov", "given": "George" } }, { "id": "Slimko-E", "name": { "family": "Slimko", "given": "Eric" } }, { "id": "Karschin-A", "name": { "family": "Karschin", "given": "Andreas" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Tyrosine decaging leads to substantial membrane trafficking during modulation of an inward rectifier potassium channel", "ispublished": "pub", "full_text_status": "public", "keywords": "unnatural amino acids, ion channel, Kir2.1, clathrin, endocytosis", "note": "\u00a9 2001 The Rockefeller University Press. \n\nSubmitted: 11 August 2000; revised: 6 December 2000; accepted: 7 December 2000. Published 16 January 2001. \n\nWe thank Hai-Rong Li for help with the oocytes, Mike Walsh for assistance with instrumentation, Tatiana Ivanina and Abraham Kovoor for advice on phosphorylation assays, and members of our laboratory for much discussion. \n\nThis research was supported by grants from the National Institutes of Health (GM29836 and NS34407), by National Research Service Awards to Y. Tong and M. Li, by a fellowship from the Burroughs-Wellcome Foundation Computational Molecular Biology Program to G. Shapovalov, by a Predoctoral Training Award from NIH to E. Slimko, by a Boehringer Fellowship to G.S. Brandt, and by the Alexander von Humboldt Foundation.\n\nPublished - TONjgp01.pdf
", "abstract": "Tyrosine side chains participate in several distinct signaling pathways, including phosphorylation and membrane trafficking. A nonsense suppression procedure was used to incorporate a caged tyrosine residue in place of the natural tyrosine at position 242 of the inward rectifier channel Kir2.1 expressed in Xenopus oocytes. When tyrosine kinases were active, flash decaging led both to decreased K+ currents and also to substantial (15\u201326%) decreases in capacitance, implying net membrane endocytosis. A dominant negative dynamin mutant completely blocked the decaging-induced endocytosis and partially blocked the decaging-induced K+ channel inhibition. Thus, decaging of a single tyrosine residue in a single species of membrane protein leads to massive clathrin-mediated endocytosis; in fact, membrane area equivalent to many clathrin-coated vesicles is withdrawn from the oocyte surface for each Kir2.1 channel inhibited. Oocyte membrane proteins were also labeled with the thiol-reactive fluorophore tetramethylrhodamine-5-maleimide, and manipulations that decreased capacitance also decreased surface membrane fluorescence, confirming the net endocytosis. In single-channel studies, tyrosine kinase activation decreased the membrane density of active Kir2.1 channels per patch but did not change channel conductance or open probability, in agreement with the hypothesis that tyrosine phosphorylation results in endocytosis of Kir2.1 channels. Despite the Kir2.1 inhibition and endocytosis stimulated by tyrosine kinase activation, neither Western blotting nor 32P labeling produced evidence for direct tyrosine phosphorylation of Kir2.1. Therefore, it is likely that tyrosine phosphorylation affects Kir2.1 function indirectly, via interactions between clathrin adaptor proteins and a tyrosine-based sorting motif on Kir2.1 that is revealed by decaging the tyrosine side chain. These interactions inhibit a fraction of the Kir2.1 channels, possibly via direct occlusion of the conduction pathway, and also lead to endocytosis, which further decreases Kir2.1 currents. These data establish that side chain decaging can provide valuable time-resolved data about intracellular signaling systems.", "date": "2001-02", "date_type": "published", "publication": "Journal of General Physiology", "volume": "117", "number": "2", "publisher": "Rockefeller University Press", "pagerange": "103-118", "id_number": "CaltechAUTHORS:TONjgp01", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:TONjgp01", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "Burroughs Wellcome Fund" }, { "agency": "Boehringer Ingelheim Fonds" }, { "agency": "Alexander von Humboldt Foundation" } ] }, "doi": "10.1085/jgp.117.2.103", "pmcid": "PMC2217249", "primary_object": { "basename": "TONjgp01.pdf", "url": "https://authors.library.caltech.edu/records/rt275-vt526/files/TONjgp01.pdf" }, "resource_type": "article", "pub_year": "2001", "author_list": "Tong, Yanhe; Brandt, Gabriel S.; et el." }, { "id": "https://authors.library.caltech.edu/records/2gax2-w7s24", "eprint_id": 103874, "eprint_status": "archive", "datestamp": "2023-08-21 21:58:11", "lastmod": "2023-10-20 16:47:50", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Chiu-Chi-Sung", "name": { "family": "Chiu", "given": "Chi-Sung" } }, { "id": "Kartalov-E-P", "name": { "family": "Kartalov", "given": "Emil" } }, { "id": "Unger-M-A", "name": { "family": "Unger", "given": "Marc" } }, { "id": "Quake-S-R", "name": { "family": "Quake", "given": "Stephen" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Single-molecule measurements calibrate green fluorescent protein surface densities on transparent beads for use with 'knock-in' animals and other expression systems", "ispublished": "pub", "full_text_status": "restricted", "keywords": "GFP (green fluorescent protein); Single-molecule imaging; Calibration; Photobleaching rate; pH sensitivity; Epi-fluorescent microscope; Histidine tag", "note": "\u00a9 2001 Elsevier. \n\nReceived 7 September 2000, Revised 30 October 2000, Accepted 30 October 2000, Available online 2 February 2001. \n\nWe thank K. Beam for the GFP37 construct, N. Dinh and M. Young (City of Hope National Medical Center) for amino acid analysis, M. Simon for providing the Nikon microscope, and N. Davidson, R. Farley, K. Philipson, and B. Khakh for discussion. This work was supported by grants from the NIH (NS-11756, DA-09121).", "abstract": "Quantitative aspects of synaptic transmission can be studied by inserting green fluorescent protein (GFP) moieties into the genes encoding membrane proteins. To provide calibrations for measurements on synapses expressing such proteins, we developed methods to quantify histidine-tagged GFP molecules (His\u2086-GFP) bound to Ni-NTA moieties on transparent beads (80\u2013120 \u03bcm diameter) over a density range comprising nearly four orders of magnitude (to 30\u2008000 GFP/\u03bcm\u00b2). The procedures employ commonly available Hg lamps, fluorescent microscopes, and CCD cameras. Two independent routes are employed: (1) single-molecule fluorescence measurements are made at the lowest GFP densities, providing an absolute calibration for macroscopic signals at higher GFP densities; (2) known numbers of His\u2086-GFP molecules are coupled quantitatively to the beads. Each of the two independent routes provides linear data over the measured density range, and the two independent methods agree with root mean square (rms) deviation of 11\u201321% over this range. These satisfactory results are obtained on two separate microscope systems. The data can be corrected for bleaching rates, which are linear with light intensity and become appreciable at intensities >\u223c1 W/cm\u00b2. If a suitable GFP-tagged protein can be chosen and incorporated into a 'knock-in' animal, the density of the protein can be measured with an absolute accuracy on the order of 20%.", "date": "2001-01-30", "date_type": "published", "publication": "Journal of Neuroscience Methods", "volume": "105", "number": "1", "publisher": "Elsevier", "pagerange": "55-63", "id_number": "CaltechAUTHORS:20200612-115646402", "issn": "0165-0270", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-115646402", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "DA-09121" } ] }, "doi": "10.1016/s0165-0270(00)00354-x", "resource_type": "article", "pub_year": "2001", "author_list": "Chiu, Chi-Sung; Kartalov, Emil; et el." }, { "id": "https://authors.library.caltech.edu/records/3xa9t-4bp59", "eprint_id": 102731, "eprint_status": "archive", "datestamp": "2023-08-21 21:55:03", "lastmod": "2023-10-20 00:27:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Lintong", "name": { "family": "Li", "given": "Lintong" } }, { "id": "Zhong-Wenge", "name": { "family": "Zhong", "given": "Wenge" } }, { "id": "Zacharias-N-M", "name": { "family": "Zacharias", "given": "Niki" } }, { "id": "Gibbs-Caroline", "name": { "family": "Gibbs", "given": "Caroline" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "The tethered agonist approach to mapping ion channel proteins \u2013 toward a structural model for the agonist binding site of the nicotinic acetylcholine receptor", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Tethered agonist; Nicotinic acetylcholine receptor; Agonist binding site; Unnatural amino acid mutagenesis", "note": "\u00a9 2001 Elsevier. Under an Elsevier user license. \n\nReceived 14 July 2000, Revised 27 October 2000, Accepted 30 October 2000, Available online 19 February 2001. \n\nESI ionization, quadrupole mass spectrometry was carried out at the Caltech Protein Microanalytical Laboratory under the direction of Gary M. Hathaway. This work was supported by the NIH (NS 34407 and NS 11756).", "abstract": "Background: The integral membrane proteins of neurons and other excitable cells are generally resistant to high resolution structural tools. Structure\u2013function studies, especially those enhanced by the nonsense suppression methodology for unnatural amino acid incorporation, constitute one of the most powerful probes of ion channels and related structures. The nonsense suppression methodology can also be used to incorporate functional side chains designed to deliver novel structural probes to membrane proteins. In this vein, we sought to generalize a potentially powerful tool \u2013 the tethered agonist approach \u2013 for mapping the agonist binding site of ligand-gated ion channels. \n\nResults: Using the in vivo nonsense suppression method for unnatural amino acid incorporation, a series of tethered quaternary ammonium derivatives of tyrosine have been incorporated into the nicotinic acetylcholine receptor. At three sites a constitutively active receptor results, but the pattern of activation as a function of chain length is different. At position \u03b1149, there is a clear preference for a three-carbon tether, while at position \u03b193 tethers of 2\u20135 carbons are comparably effective. At position \u03b355/\u03b457 all tethers except the shortest one can activate the receptor. Based on these and other data, a model for the receptor binding site can be developed by analogy to the acetylcholine esterase crystal structure. \n\nConclusion: Through the use of nonsense suppression techniques, the tethered agonist approach has been made into a general tool for probing receptor structures. When applied to the nicotinic receptor, the method places new restrictions on developing models for the agonist binding site.", "date": "2001-01", "date_type": "published", "publication": "Chemistry and Biology", "volume": "8", "number": "1", "publisher": "Elsevier", "pagerange": "47-58", "id_number": "CaltechAUTHORS:20200422-150959147", "issn": "1074-5521", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200422-150959147", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1016/s1074-5521(00)00055-7", "resource_type": "article", "pub_year": "2001", "author_list": "Li, Lintong; Zhong, Wenge; et el." }, { "id": "https://authors.library.caltech.edu/records/96znm-gym07", "eprint_id": 13265, "eprint_status": "archive", "datestamp": "2023-08-21 21:45:13", "lastmod": "2023-10-17 23:22:08", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Chiu-Chi-Sung", "name": { "family": "Chiu", "given": "Chi-Sung" } }, { "id": "Ross-L-S", "name": { "family": "Ross", "given": "Linda S." } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Gill-S-S", "name": { "family": "Gill", "given": "Sarjeet S." } } ] }, "title": "The transporter-like protein inebriated mediates hyperosmotic stimuli through intracellular signaling", "ispublished": "pub", "full_text_status": "public", "keywords": "Manduca sexta; inebriated; Ca2+-activated Cl- current; inositol trisphosphate; permeable phospholipase C", "note": "\u00a9 The Company of Biologists Limited 2000. \n\nAccepted 7 September; published on WWW 2 November 2000. \n\nWe thank M. E. Adams, C. Y. Lytle, M. J. Schuler, D. Zitnan, A. K. Pullikuth, C. Labarca, M. Filippova, A. Kondo and L. McCloud for helpful discussion and technical support. This research was supported by grants from the NIH (AI34524 and AI48049 to S.S.G. and DA-09121 to H.A.L.).\n\nPublished - CHIjeb00.pdf
", "abstract": "We cloned the inebriated homologue MasIne from Manduca sexta and expressed it in Xenopus laevis oocytes. MasIne is homologous to neurotransmitter transporters but no transport was observed with a number of putative substrates. Oocytes expressing MasIne respond to hyperosmotic stimulation by releasing intracellular Ca(2+), as revealed by activation of the endogenous Ca(2+)-activated Cl(-) current. This Ca(2+) release requires the N-terminal 108 amino acid residues of MasIne and occurs via the inositol trisphosphate pathway. Fusion of the N terminus to the rat gamma-aminobutyric acid transporter (rGAT1) also renders rGAT1 responsive to hyperosmotic stimulation. Immunohistochemical analyses show that MasIne and Drosophila Ine have similar tissue distribution patterns, suggesting functional identity. Inebriated is expressed in tissues and cells actively involved in K(+) transport, which suggests that it may have a role in ion transport, particularly of K(+). We propose that stimulation of MasIne releases intracellular Ca(2+) in native tissues, activating Ca(2+)-dependent K(+) channels, and leading to K(+) transport.", "date": "2000-12-01", "date_type": "published", "publication": "Journal of Experimental Biology", "volume": "203", "number": "23", "publisher": "Company of Biologists", "pagerange": "3531-3546", "id_number": "CaltechAUTHORS:CHIjeb00", "issn": "0022-0949", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:CHIjeb00", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "AI4524" }, { "agency": "NIH", "grant_number": "AI48049" }, { "agency": "NIH", "grant_number": "DA-09121" } ] }, "primary_object": { "basename": "CHIjeb00.pdf", "url": "https://authors.library.caltech.edu/records/96znm-gym07/files/CHIjeb00.pdf" }, "resource_type": "article", "pub_year": "2000", "author_list": "Chiu, Chi-Sung; Ross, Linda S.; et el." }, { "id": "https://authors.library.caltech.edu/records/mfr1v-ag708", "eprint_id": 103876, "eprint_status": "archive", "datestamp": "2023-08-21 21:43:17", "lastmod": "2023-10-20 16:48:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Ehrengruber-M-U", "name": { "family": "Ehrengruber", "given": "Markus U." } }, { "id": "Muhlebach-S-G", "name": { "family": "Muhlebach", "given": "Stephan G." } }, { "id": "S\u00f6hrman-S", "name": { "family": "S\u00f6hrman", "given": "Sophia" } }, { "id": "Leutenegger-C-M", "name": { "family": "Leutenegger", "given": "Christian M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Modulation of early growth response (EGR) transcription factor-dependent gene expression by using recombinant adenovirus", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Immediate early gene; Myoblast; Neuroblastoma; NGFI-A binding protein 2 (NAB2); Quantitative PCR; Viral gene transfer", "note": "\u00a9 2000 Elsevier. \n\nReceived 16 June 2000, Revised 15 September 2000, Accepted 28 September 2000, Available online 4 December 2000. \n\nWe thank Dr Jeffrey Milbrandt and Dr John Svaren for providing A2ProLuc and the cDNAs for EGR1, EGR1\u2217, EGR2, NAB2, R1, and R1\u2217, Dr Barbara J. Wold and Dr Jeong Kyo Yoon for the sGFP cDNA, Anita Buchli for the Neuro-2a cells, Susanne Erb for the C2C12 cells, Dr Hanns M\u00f6hler and Laura Huopaniemi for supporting the TaqMan PCR, and Dr John Svaren and Dr Sondra Schlesinger for helpful discussions and comments on the manuscript. This work was supported by the Swiss National Science Foundation (grant no. 31-57\u2032125.99 to MUE, and fellowship no. 823A-053469 to CML), the National Institute of Mental Health (grant to ND), and by the Donald E. and Delia B. Baxter Foundation (fellowship to MUE).", "abstract": "Early growth response (EGR) transcription factors link initial cytoplasmic events to long-term alterations of cellular gene expression and are induced by various stimuli. To test their roles in cell physiology, we constructed adenoviral recombinants encoding NGFI-A binding protein 2 (NAB2, a repressor of EGR1, EGR2, and EGR3), EGR1, NAB-insensitive EGR1(I293F) (EGR1\u2217), EGR2, and the NAB-binding, repressive domain 1 (R1) of EGR1. These viruses regulated EGR-dependent expression of GFP and luciferase reporter genes in heterologous expression assays. Infection of a myoblast cell line with EGR1 and EGR1\u2217 adenovirus induced the endogenous gene for platelet-derived growth factor A chain (PDGF-A). In addition, in neuroblastoma cells, the two novel EGR1 target genes EGR3 and NAB2 were identified by using adenoviral transfer of EGR1 and EGR1\u2217. Our results demonstrate that recombinant adenovirus is useful to regulate heterologous and endogenous EGR target gene expression, and suggest that EGR transcription factors can autoregulate themselves.", "date": "2000-11-27", "date_type": "published", "publication": "Gene", "volume": "258", "number": "1-2", "publisher": "Elsevier", "pagerange": "63-69", "id_number": "CaltechAUTHORS:20200612-115646596", "issn": "0378-1119", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-115646596", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Swiss National Science Foundation (SNSF)", "grant_number": "31-57\u2032125.99" }, { "agency": "Swiss National Science Foundation (SNSF)", "grant_number": "823A-053469" }, { "agency": "National Institute of Mental Health (NIMH)" }, { "agency": "NIH" }, { "agency": "Donald E. and Delia B. Baxter Foundation" } ] }, "doi": "10.1016/s0378-1119(00)00445-5", "resource_type": "article", "pub_year": "2000", "author_list": "Ehrengruber, Markus U.; Muhlebach, Stephan G.; et el." }, { "id": "https://authors.library.caltech.edu/records/n9dge-qtw57", "eprint_id": 103879, "eprint_status": "archive", "datestamp": "2023-08-19 06:10:31", "lastmod": "2023-10-20 16:48:22", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kofuji-Paulo", "name": { "family": "Kofuji", "given": "Paulo" } }, { "id": "Ceelen-P", "name": { "family": "Ceelen", "given": "Paul" } }, { "id": "Zahs-K-R", "name": { "family": "Zahs", "given": "Kathleen R." } }, { "id": "Surbeck-L-W", "name": { "family": "Surbeck", "given": "Leslie W." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Newman-E-A", "name": { "family": "Newman", "given": "Eric A." } } ] }, "title": "Genetic Inactivation of an Inwardly Rectifying Potassium Channel (Kir4.1 Subunit) in Mice: Phenotypic Impact in Retina", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2000 Society for Neuroscience. \n\nReceived April 4, 2000; revised May 12, 2000; accepted May 17, 2000. \n\nThis work was supported by National Institute of Health Grants GM-29836, MH-49176, EY04077, EY10383, and EY12949. We thank S. Pease, M. Larabee, and T. Wu for expert technical help.\n\nPublished - 5733.full.pdf
", "abstract": "The inwardly rectifying potassium channel Kir4.1 has been suggested to underlie the principal K\u207a conductance of mammalian M\u00fcller cells and to participate in the generation of field potentials and regulation of extracellular K\u207a in the retina. To further assess the role of Kir4.1 in the retina, we generated a mouse line with targeted disruption of the Kir4.1 gene (Kir4.1 \u2212/\u2212). M\u00fcller cells from Kir4.1 \u2212/\u2212 mice were not labeled with an anti-Kir4.1 antibody, although they appeared morphologically normal when stained with an anti-glutamine synthetase antibody. In contrast, in M\u00fcller cells from wild-type littermate (Kir4.1 +/+) mice, Kir4.1 was present and localized to the proximal endfeet and perivascular processes. In situ whole-cell patch-clamp recordings showed a 10-fold increase in the input resistance and a large depolarization of Kir4.1 \u2212/\u2212 M\u00fcller cells compared with Kir4.1 +/+ cells. The slow PIII response of the light-evoked electroretinogram (ERG), which is generated by K\u207a fluxes through M\u00fcller cells, was totally absent in retinas from Kir4.1 \u2212/\u2212 mice. The b-wave of the ERG, in contrast, was spared in the null mice. Overall, these results indicate that Kir4.1 is the principal K\u207a channel subunit expressed in mouse M\u00fcller glial cells. The highly regulated localization and the functional properties of Kir4.1 in M\u00fcller cells suggest the involvement of this channel in the regulation of extracellular K\u207a in the mouse retina.", "date": "2000-08-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "20", "number": "15", "publisher": "Society for Neuroscience", "pagerange": "5733-5740", "id_number": "CaltechAUTHORS:20200612-132259782", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-132259782", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "EY04077" }, { "agency": "NIH", "grant_number": "EY10383" }, { "agency": "NIH", "grant_number": "EY12949" } ] }, "doi": "10.1523/jneurosci.20-15-05733.2000", "pmcid": "PMC2410027", "primary_object": { "basename": "5733.full.pdf", "url": "https://authors.library.caltech.edu/records/n9dge-qtw57/files/5733.full.pdf" }, "resource_type": "article", "pub_year": "2000", "author_list": "Kofuji, Paulo; Ceelen, Paul; et el." }, { "id": "https://authors.library.caltech.edu/records/frn9z-bgz33", "eprint_id": 55585, "eprint_status": "archive", "datestamp": "2023-08-19 06:08:49", "lastmod": "2023-10-20 22:24:40", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nadeau-H", "name": { "family": "Nadeau", "given": "H." } }, { "id": "Mckinney-S", "name": { "family": "Mckinney", "given": "S." } }, { "id": "Anderson-D-J", "name": { "family": "Anderson", "given": "D. J." }, "orcid": "0000-0001-6175-3872" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "ROMK1 (Kir1.1) Causes Apoptosis and Chronic Silencing of Hippocampal Neurons", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2000 The American Physiological Society.\n\nSubmitted 27 March 2000; accepted in final form 4 May 2000.\n\nWe thank B. Khakh, G. Greif, J. Pine, and C. Lindensmith for useful suggestions and discussions. This work was supported by Burroughs-Wellcome and by National Institute of Mental Health Grant MH-49176.", "abstract": "Lentiviral vectors were constructed to express the weakly rectifying kidney K^+ channel ROMK1 (Kir1.1), either fused to enhanced green fluorescent protein (EGFP) or as a bicistronic message (ROMK1-CITE-EGFP). The channel was stably expressed in cultured rat hippocampal neurons. Infected cells were maintained for 2\u20134 wk without decrease in expression level or evidence of viral toxicity, although 15.4 mM external KCl was required to prevent apoptosis of neurons expressing functional ROMK1. No other trophic agents tested could prevent cell death, which was probably caused by K+ loss. This cell death did not occur in glia, which were able to support ROMK1 expression indefinitely. Functional ROMK1, quantified as the nonnative inward current at \u2212144 mV in 5.4 mM external K+blockable by 500 \u03bcM Ba^2+, ranged from 1 to 40 pA/pF. Infected neurons exhibited a Ba^2+-induced depolarization of 7 \u00b1 2 mV relative to matched EGFP-infected controls, as well as a 30% decrease in input resistance and a shift in action potential threshold of 2.6 \u00b1 0.5 mV. This led to a shift in the relation between injected current and firing frequency, without changes in spike shape, size, or timing. This shift, which quantifies silencing as a function of ROMK1 expression, was predicted from Hodgkin-Huxley models. No cellular compensatory mechanisms in response to expression of ROMK1 were identified, making ROMK1 potentially useful for transgenic studies of silencing and neurodegeneration, although its lethality in normal K^+ has implications for the use of K^+ channels in gene therapy.", "date": "2000-08", "date_type": "published", "publication": "Journal of Neurophysiology", "volume": "84", "number": "2", "publisher": "American Physiological Society", "pagerange": "1062-1075", "id_number": "CaltechAUTHORS:20150306-095241138", "issn": "0022-3077", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150306-095241138", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Burroughs-Wellcome Fund" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "National Institute of Mental Health (NIMH)" } ] }, "doi": "10.1152/jn.2000.84.2.1062", "resource_type": "article", "pub_year": "2000", "author_list": "Nadeau, H.; Mckinney, S.; et el." }, { "id": "https://authors.library.caltech.edu/records/a25g2-g3g62", "eprint_id": 56190, "eprint_status": "archive", "datestamp": "2023-08-19 06:07:42", "lastmod": "2023-10-23 15:08:24", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Khakh-B-S", "name": { "family": "Khakh", "given": "Baljit S." } }, { "id": "Zhou-Xiaoping", "name": { "family": "Zhou", "given": "Xiaoping" } }, { "id": "Sydes-J", "name": { "family": "Sydes", "given": "Jason" } }, { "id": "Galligan-J-J", "name": { "family": "Galligan", "given": "James J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "State-dependent cross-inhibition between transmitter-gated cation channels", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2000 Macmillan Magazines Ltd.\n\nReceived 12 April; accepted 26 May 2000.\n\nThanks to H. Li for assistance with preparation of oocytes, and to other members of the group for comments. A Wellcome Trust (UK) International Prize Travelling Fellowship (to B.S.K.) and the NIH supported this work.\n\nSupplemental Material - fig1.pdf
Supplemental Material - fig2.pdf
Supplemental Material - fig3.pdf
Supplemental Material - fig4.pdf
Supplemental Material - table.doc
", "abstract": "Transmitter-gated cation channels are detectors of excitatory chemical signals at synapses in the nervous system. Here we show that structurally distinct \u03b13\u03b24 nicotinic and P2X_2 channels influence each other when co-activated. The activation of one channel type affects distinct kinetic and conductance states of the other, and co-activation results in non-additive responses owing to inhibition of both channel types. State-dependent inhibition of nicotinic channels is revealed most clearly with mutant P2X_2 channels, and inhibition is decreased at lower densities of channel expression. In synaptically coupled myenteric neurons, nicotinic fast excitatory postsynaptic currents are occluded during activation of endogenously co-expressed P2X channels. Our data provide a molecular basis and a synaptic context for cross-inhibition between transmitter-gated channels.", "date": "2000-07-27", "date_type": "published", "publication": "Nature", "volume": "406", "number": "6794", "publisher": "Nature Publishing Group", "pagerange": "405-410", "id_number": "CaltechAUTHORS:20150327-133158044", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150327-133158044", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "NIH" } ] }, "doi": "10.1038/35019066", "primary_object": { "basename": "fig2.pdf", "url": "https://authors.library.caltech.edu/records/a25g2-g3g62/files/fig2.pdf" }, "related_objects": [ { "basename": "fig3.pdf", "url": "https://authors.library.caltech.edu/records/a25g2-g3g62/files/fig3.pdf" }, { "basename": "fig4.pdf", "url": "https://authors.library.caltech.edu/records/a25g2-g3g62/files/fig4.pdf" }, { "basename": "table.doc", "url": "https://authors.library.caltech.edu/records/a25g2-g3g62/files/table.doc" }, { "basename": "fig1.pdf", "url": "https://authors.library.caltech.edu/records/a25g2-g3g62/files/fig1.pdf" } ], "resource_type": "article", "pub_year": "2000", "author_list": "Khakh, Baljit S.; Zhou, Xiaoping; et el." }, { "id": "https://authors.library.caltech.edu/records/6grps-m7651", "eprint_id": 5103, "eprint_status": "archive", "datestamp": "2023-08-21 21:23:53", "lastmod": "2023-10-16 18:07:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Maurer-J-A", "name": { "family": "Maurer", "given": "Joshua A." } }, { "id": "Elmore-D-E", "name": { "family": "Elmore", "given": "Donald E." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Comparing and contrasting Escherichia coli and Mycobacterium tuberculosis mechanosensitive channels (MscL) - New gain of function mutations in the loop region", "ispublished": "pub", "full_text_status": "public", "keywords": "LARGE-CONDUCTANCE; SINGLE RESIDUE; ION-CHANNEL; PROTEIN; ALIGNMENT", "note": "\u00a9 2000 by the American Society for Biochemistry and Molecular Biology. \n\nReceived for publication, April 11, 2000, and in revised form, May 5, 2000. http://dx.doi.org/10.1109/50.933286 \n\nWe are grateful to Prof. Douglas Rees, Dr. Randal Bass, Dr. Geoffrey Chang, Dr. Robert Spencer, and George Shapovalov for helpful discussions. Plasmids and wild type Tb-MscL proteins used in these studies were obtained from the Rees group. MALDI-TOF mass analyses were carried out at the Caltech Protein Microanalytical Laboratory under the direction of Gary M. Hathaway.\n\nPublished - MAUjbc00.pdf
", "abstract": "Sequence analysis of 35 putative MscL homologues was used to develop an optimal alignment for Escherichia coli and Mycobacterium tuberculosis MscL and to place these homologues into sequence subfamilies. By using this alignment, previously identified E. coli MscL mutants that displayed severe and very severe gain of function phenotypes were mapped onto the M. tuberculosis MscL sequence. Not all of the resulting M. tuberculosis mutants displayed a gain of function phenotype; for instance, normal phenotypes were noted for mutations at Ala20, the analogue of the highly sensitive Gly22 site in E. coli. A previously unnoticed intersubunit hydrogen bond in the extracellular loop region of the M. tuberculosis MscL crystal structure has been analyzed. Cross-linkable residues were substituted for the residues involved in the hydrogen bond, and cross-linking studies indicated that these sites are spatially close under physiological conditions. In general, mutation at these positions results in a gain of function phenotype, which provides strong evidence for the importance of the loop region in MscL channel function. No analogue to this interesting interaction could be found in E. coli MscL by sequence alignment. Taken together, these results indicate that caution should be exercised in using the M. tuberculosis MscL crystal structure to analyze previous functional studies of E. coli MscL.", "date": "2000-07-21", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "275", "number": "29", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "22238-22244", "id_number": "CaltechAUTHORS:MAUjbc00", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:MAUjbc00", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1074/jbc.M003056200", "primary_object": { "basename": "MAUjbc00.pdf", "url": "https://authors.library.caltech.edu/records/6grps-m7651/files/MAUjbc00.pdf" }, "resource_type": "article", "pub_year": "2000", "author_list": "Maurer, Joshua A.; Elmore, Donald E.; et el." }, { "id": "https://authors.library.caltech.edu/records/jnsmk-zwf16", "eprint_id": 91759, "eprint_status": "archive", "datestamp": "2023-08-21 21:18:49", "lastmod": "2023-10-19 23:43:08", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nadeau-H", "name": { "family": "Nadeau", "given": "H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Two-compartment model for whole-cell data analysis and transient compensation", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Whole-cell recording; Series resistance; Compensation; Capacitance; Compartmental model; Cable; Voltage-clamp", "note": "\u00a9 2000 Elsevier.\n\nReceived 12 December 1999, Revised 20 March 2000, Accepted 20 March 2000, Available online 7 August 2000.", "abstract": "Recording and analysis of neuronal patch-clamp data involve many assumptions about membrane properties and cell morphology. Some of these assumptions introduce large errors or oversimplifications into the results. In particular, dendritic branching with high intracellular resistance leads to difficulty with capacitance calculation and transient subtraction, and may significantly distort measured currents. A two-compartment model, presented in detail here, provides a simple method of reducing many of these problems for the relatively simple case of cultured neurons studied with whole-cell patch electrodes. Some passive membrane properties may be accurately calculated, and the results may be used to correct recorded currents for resulting series resistance, intracellular resistance, and capacitive transient errors. The model may be tailored to particular cell types or experimental conditions. Programs to implement the algorithms are available from http://www.its.caltech.edu/\u223cnadeau/Rscomp.html.", "date": "2000-06-30", "date_type": "published", "publication": "Journal of Neuroscience Methods", "volume": "99", "number": "1-2", "publisher": "Elsevier", "pagerange": "25-35", "id_number": "CaltechAUTHORS:20181213-110051972", "issn": "0165-0270", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20181213-110051972", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1016/s0165-0270(00)00210-7", "resource_type": "article", "pub_year": "2000", "author_list": "Nadeau, H. and Lester, H. A." }, { "id": "https://authors.library.caltech.edu/records/czkhs-m8v23", "eprint_id": 103161, "eprint_status": "archive", "datestamp": "2023-08-19 05:54:09", "lastmod": "2023-10-20 15:43:14", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dang-Hong", "name": { "family": "Dang", "given": "Hong" } }, { "id": "England-P-M", "name": { "family": "England", "given": "Pamela M." } }, { "id": "Farivar-S-S", "name": { "family": "Farivar", "given": "S. Sarah" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Probing the Role of a Conserved M1 Proline Residue in 5-Hydroxytryptamine\u2083 Receptor Gating", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 2000 The American Society for Pharmacology and Experimental Therapeutics. \n\nReceived December 9, 1999; accepted February 12, 2000. \n\nWe thank Justin Gallivan, Scott Silverman, and Wenge Zhong for sharing synthetic dCA amino acid analogs and Hairong Li for preparingX. laevis oocytes used in our experiments. We also thank Drs. D. Julius (UCSF) and E. Kirkness (TIGR) for the 5-HT\u2083 cDNA clones and Dr. P. Schultz for P3m. \n\nThis work was supported by National Institutes of Health Grants MH49176, NS34407, and NS10305.", "abstract": "A conserved proline residue is found in the first transmembrane domain (M1) of every subunit in the ligand-gated ion channel superfamily. The position of this proline between the N-terminal extracellular agonist binding and the second transmembrane (M2) channel lining domains in the primary sequence suggests its possible involvement in the gating of the receptor. Replacing this proline with alanine, glycine, or leucine in the 5-hydroxytryptamine (5-HT)_(3A) homomeric receptors expressed in Xenopus laevis oocytes resulted in the absence of 5-HT-induced whole-cell currents, although there were normal levels of specific surface [\u00b3H]granisetron ([\u00b3H]BRL-43694) binding sites. To determine what properties of the conserved proline are critical for the function of the channel, two imino acids and an \u03b1-hydroxy acid were incorporated at the proline position using the nonsense suppression method.trans-3-Methyl-proline, pipecolic acid, and leucic acid were able to replace the conserved proline to produce active channels with EC\u2085\u2080 values similar to that for the wild-type receptor. These trends are preserved in the heteromeric receptors consisting of 5-HT_(3A) and 5-HT_(3B) subunits in oocytes. The prominent common feature among these residues and proline is the lack of hydrogen bond donor activity, potentially resulting in a flexible secondary structure in the M1 region. Thus, lack of hydrogen bond donor activity may be a key element in channel gating and may explain the high degree of conservation of this M1 proline.", "date": "2000-06-01", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "57", "number": "6", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "1114-1122", "id_number": "CaltechAUTHORS:20200513-070503095", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200513-070503095", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "MH49176" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "NS10305" } ] }, "resource_type": "article", "pub_year": "2000", "author_list": "Dang, Hong; England, Pamela M.; et el." }, { "id": "https://authors.library.caltech.edu/records/1aqxz-y5j81", "eprint_id": 6577, "eprint_status": "archive", "datestamp": "2023-08-21 21:02:32", "lastmod": "2023-10-16 20:24:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Ming", "name": { "family": "Li", "given": "Ming" } }, { "id": "Farley-R-A", "name": { "family": "Farley", "given": "Robert A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "An Intermediate State of the {gamma}-Aminobutyric Acid Transporter GAT1 Revealed by Simultaneous Voltage Clamp and Fluorescence", "ispublished": "pub", "full_text_status": "public", "keywords": "voltage clamp; Xenopus oocyte; tetramethylrhodamine; conformational change", "note": "\u00a9 2000 The Rockefeller University Press \n\nSubmitted: 7 December 1999; Revised: 11 February 2000; Accepted: 22 February 2000; Published 27 March 2000. doi:10.1085/jgp.115.4.491 \n\nWe thank Mike Walsh for excellent technical assistance and Ehud Isacoff, Micah Siegel, and Yong-Xin Li for advice and reagents. \n\nThis work is supported by grants from the National Institutes of Health (NS-11756, DA-09121).\n\nPublished - LIMjgp00.pdf
", "abstract": "The rat {gamma}-aminobutyric acid transporter GAT1 expressed in Xenopus oocytes was labeled at Cys74, and at one or more other sites, by tetramethylrhodamine-5-maleimide, without significantly altering GAT1 function. Voltage-jump relaxation analysis showed that fluorescence increased slightly and monotonically with hyperpolarization; the fluorescence at -140 mV was ~0.8% greater than at +60 mV. The time course of the fluorescence relaxations was mostly described by a single exponential with voltage-dependent but history-independent time constants ranging from ~20 ms at +60 mV to ~150 ms at -140 mV. The fluorescence did not saturate at the most negative potentials tested, and the midpoint of the fluorescence\u2013voltage relation was at least 50 mV more negative than the midpoint of the charge\u2013voltage relation previously identified with Na+ binding to GAT1. The presence of {gamma}-aminobutyric acid did not noticeably affect the fluorescence waveforms. The fluorescence signal depended on Na+ concentration with a Hill coefficient approaching 2. Increasing Cl- concentration modestly increased and accelerated the fluorescence relaxations for hyperpolarizing jumps. The fluorescence change was blocked by the GAT1 inhibitor, NO-711. For the W68L mutant of GAT1, the fluorescence relaxations occurred only during jumps to high positive potentials, in agreement with previous suggestions that this mutant is trapped in one conformational state except at these potentials. These observations suggest that the fluorescence signals monitor a novel state of GAT1, intermediate between the E*out and Eout states of Hilgemann, D.W., and C.-C. Lu (1999. J. Gen. Physiol. 114:459\u2013476). Therefore, the study provides verification that conformational changes occur during GAT1 function.", "date": "2000-04", "date_type": "published", "publication": "Journal of General Physiology", "volume": "115", "number": "4", "publisher": "Rockefeller University Press", "pagerange": "491-508", "id_number": "CaltechAUTHORS:LIMjgp00", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LIMjgp00", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "DA-09121" } ] }, "doi": "10.1085/jgp.115.4.491", "pmcid": "PMC2233755", "primary_object": { "basename": "LIMjgp00.pdf", "url": "https://authors.library.caltech.edu/records/1aqxz-y5j81/files/LIMjgp00.pdf" }, "resource_type": "article", "pub_year": "2000", "author_list": "Li, Ming; Farley, Robert A.; et el." }, { "id": "https://authors.library.caltech.edu/records/a4r9k-2wf20", "eprint_id": 1458, "eprint_status": "archive", "datestamp": "2023-08-21 20:56:40", "lastmod": "2023-10-13 22:49:44", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Karschin-A", "name": { "family": "Karschin", "given": "Andreas" } } ] }, "title": "Gain of function mutants: Ion channels and G protein-coupled receptors", "ispublished": "pub", "full_text_status": "public", "keywords": "EPILEPSY; NEURODEGENERATION; WEAVER; GENETIC DISEASE; LONG-QT SYNDROME; EPITHELIAL NA+ CHANNEL; KNOCKDOWN-RESISTANCE KDR; FRONTAL-LOBE EPILEPSY; INHERITED CARDIAC-ARRHYTHMIA; RECTIFYING K+ CHANNELS; GATED SODIUM-CHANNEL; CONSTITUTIVELY ACTIVATING MUTATION; COCKROACHES BLATTELLA-GERMANICA; NEMATODE CAENORHABDITIS-ELEGANS", "note": "\"Reprinted, with permission, from the Annual Review of Neuroscience, Volume 23 copyright 2000 by Annual Reviews, www.annualreviews.org\" \n\nWe thank A Auerbach, NJM Birdsall, J Garcia-Anoveros, R Dingledine, AG Engel, F Lehmann-Horn, D Mackinnon, JL Noebels, RL Ruff, M Sanguinetti, W Shi, M Yuzaki, and our colleagues at Caltech and Gottingen for many comments. Preparation of this chapter was supported by the Alexander von Humboldt Foundation. Work in our laboratories on this topic is supported by National Institutes of Health grants GM29836, MH49176, and NS11756, by the California Tobacco-Related Disease Research Program, and by the Sidney Stern Foundation.\n\nPublished - LESarn00.pdf
", "abstract": "Many ion channels and receptors display striking phenotypes for gain-of-function mutations but milder phenotypes for null mutations. Gain of molecular function can have several mechanistic bases: selectivity changes, gating changes including constitutive activation and slowed inactivation, elimination of a subunit that enhances inactivation, decreased drug sensitivity, changes in regulation or trafficking of the channel, or induction of apoptosis. Decreased firing frequency can occur via increased function of K+ or Cl- channels. Channel mutants also cause gain-of-function syndromes at the cellular and circuit level; of these syndromes, the cardiac long-QT syndromes are explained in a more straightforward way than are the epilepsies. G protein-coupled receptors are also affected by activating mutations.", "date": "2000-03", "date_type": "published", "publication": "Annual Review of Neuroscience", "volume": "23", "publisher": "Annual Reviews", "pagerange": "89-125", "id_number": "CaltechAUTHORS:LESarn00", "issn": "0147-006X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LESarn00", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Alexander von Humboldt Foundation" }, { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "MH49176" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Sidney Stern Foundation" } ] }, "doi": "10.1146/annurev.neuro.23.1.89", "primary_object": { "basename": "LESarn00.pdf", "url": "https://authors.library.caltech.edu/records/a4r9k-2wf20/files/LESarn00.pdf" }, "resource_type": "article", "pub_year": "2000", "author_list": "Lester, Henry A. and Karschin, Andreas" }, { "id": "https://authors.library.caltech.edu/records/sfvp9-g0t27", "eprint_id": 7620, "eprint_status": "archive", "datestamp": "2023-08-21 20:54:19", "lastmod": "2023-10-16 21:00:24", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kovoor-A", "name": { "family": "Kovoor", "given": "Abraham" } }, { "id": "Chen-Ching-Kang", "name": { "family": "Chen", "given": "Ching-Kang" } }, { "id": "He-Wei", "name": { "family": "He", "given": "Wei" } }, { "id": "Wensel-T-G", "name": { "family": "Wensel", "given": "Theodore G." } }, { "id": "Simon-M-I", "name": { "family": "Simon", "given": "Melvin I." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Co-expression of Gbeta 5 Enhances the Function of Two Ggamma Subunit-like Domain-containing Regulators of G Protein Signaling Proteins", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 2000 by The American Society for Biochemistry and Molecular Biology, Inc. \n\n(Received for publication, August 5, 1999, and in revised form, October 21, 1999) \n\nThe costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.\n\nPublished - KOVjbc00.pdf
", "abstract": "Regulators of G protein signaling (RGS) stimulate the GTPase activity of G protein Galpha subunits and probably play additional roles. Some RGS proteins contain a Ggamma subunit-like (GGL) domain, which mediates a specific interaction with Gbeta 5. The role of such interactions in RGS function is unclear. RGS proteins can accelerate the kinetics of coupling of G protein-coupled receptors to G-protein-gated inwardly rectifying K+ (GIRK) channels. Therefore, we coupled m2-muscarinic acetylcholine receptors to GIRK channels in Xenopus oocytes to evaluate the effect of Gbeta 5 on RGS function. Co-expression of either RGS7 or RGS9 modestly accelerated GIRK channel kinetics. When Gbeta 5 was co-expressed with either RGS7 or RGS9, the acceleration of GIRK channel kinetics was strongly increased over that produced by RGS7 or RGS9 alone. RGS function was not enhanced by co-expression of Gbeta 1, and co-expression of Gbeta 5 alone had no effect on GIRK channel kinetics. Gbeta 5 did not modulate the function either of RGS4, an RGS protein that lacks a GGL domain, or of a functional RGS7 construct in which the GGL domain was omitted. Enhancement of RGS7 function by Gbeta 5 was not a consequence of an increase in the amount of plasma membrane or cytosolic RGS7 protein.", "date": "2000-02-04", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "275", "number": "5", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "3397-3402", "id_number": "CaltechAUTHORS:KOVjbc00", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:KOVjbc00", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "primary_object": { "basename": "KOVjbc00.pdf", "url": "https://authors.library.caltech.edu/records/sfvp9-g0t27/files/KOVjbc00.pdf" }, "resource_type": "article", "pub_year": "2000", "author_list": "Kovoor, Abraham; Chen, Ching-Kang; et el." }, { "id": "https://authors.library.caltech.edu/records/p76ek-b1033", "eprint_id": 103880, "eprint_status": "archive", "datestamp": "2023-08-21 20:52:26", "lastmod": "2023-10-20 16:48:30", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Ivanina-T", "name": { "family": "Ivanina", "given": "Tatiana" } }, { "id": "Neusch-C", "name": { "family": "Neusch", "given": "Clemens" } }, { "id": "Li-Yong-Xin", "name": { "family": "Li", "given": "Yong-Xin" } }, { "id": "Tong-Yanhe", "name": { "family": "Tong", "given": "Yanhe" } }, { "id": "Labarca-C-G", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Mosher-D-F", "name": { "family": "Mosher", "given": "Deane F." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Expression of GIRK (Kir3.1/Kir3.4) channels in mouse fibroblast cells with and without \u03b21 integrins", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Voltage clamp; Inward rectifier K channel; Fibroblast; Green fluorescent protein", "note": "\u00a9 2000 Federation of European Biochemical Societies. \n\nReceived 22 November 1999. \n\nWe thank Nathan Dascal for helpful discussion, Steve Potter for help with confocal microscopy, and Hai\u2010Rong Li for help with the oocytes. This work was supported by grants from the National Institutes of Health (GM\u201029836, HL21644), the US\u2010Israel Binational Science Foundation, and the Deutsche Forschungsgemeinschaft.", "abstract": "G protein\u2010activated K\u207a channel (GIRK) subunits possess a conserved extracellular integrin\u2010binding motif (RGD) and bind directly to \u03b21 integrins. We expressed GIRK1/GIRK4 channels labeled with green fluorescent protein in fibroblast cell lines expressing or lacking \u03b21 integrins. Neither plasma membrane localization nor agonist\u2010evoked GIRK currents were affected by the absence of \u03b21 integrins or by incubation with externally applied RGD\u2010containing peptide. Mutation of the aspartate (D) of RGD impaired currents, GIRK glycosylation, and membrane localization, but the interaction with \u03b21 integrins remained intact. Thus, \u03b21 integrins are not essential for functional GIRK expression; and the GIRK\u2010integrin interactions involve structural elements other than the RGD motif.", "date": "2000-01-28", "date_type": "published", "publication": "FEBS Letters", "volume": "466", "number": "2-3", "publisher": "Federation of European Biochemical Societies", "pagerange": "327-332", "id_number": "CaltechAUTHORS:20200612-132259910", "issn": "0014-5793", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-132259910", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM\u201029836" }, { "agency": "NIH", "grant_number": "HL21644" }, { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)" } ] }, "doi": "10.1016/s0014-5793(99)01738-x", "resource_type": "article", "pub_year": "2000", "author_list": "Ivanina, Tatiana; Neusch, Clemens; et el." }, { "id": "https://authors.library.caltech.edu/records/pxydn-wpc12", "eprint_id": 103881, "eprint_status": "archive", "datestamp": "2023-08-19 04:51:18", "lastmod": "2023-10-20 16:48:33", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Unger-M-A", "name": { "family": "Unger", "given": "M." } }, { "id": "Kartalov-E-P", "name": { "family": "Kartalov", "given": "E." } }, { "id": "Chiu-Chi-Sung", "name": { "family": "Chiu", "given": "C.-S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Quake-S-R", "name": { "family": "Quake", "given": "S. R." } } ] }, "title": "Single-Molecule Fluorescence Observed with Mercury Lamp Illumination", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1999 Future Science Group. \n\nReceived 11 February 1999; accepted 13 July 1999. \n\nThe S65T, V163A, I167T, S175G GFP mutant was kindly provided by Dr. Kurt Beam, Department of Anatomy and Neurobiology, Colorado State University, Fort Collins, CO, USA. This work was partially supported by the National Institutes of Health Grant Nos. NS-11756 and DA-9121.\n\nPublished - 99275rr01.pdf
", "abstract": "We demonstrate that it is possible to observe single fluorescent molecules using a standard fluorescence microscope with mercury lamp excitation and an inexpensive cooled charge-coupled device (CCD) camera. With this equipment, we have been able to observe single molecules of tetramethyl-rhodamine, rhodamine 6G, fluorescein isothiocyanate and green fluorescent protein. Immobilized molecules were observed both in air and in aqueous solution.", "date": "1999-11", "date_type": "published", "publication": "BioTechniques", "volume": "27", "number": "5", "publisher": "Informa Healthcare", "pagerange": "1008-1014", "id_number": "CaltechAUTHORS:20200612-132300221", "issn": "0736-6205", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200612-132300221", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "DA-9121" } ] }, "doi": "10.2144/99275rr01", "primary_object": { "basename": "99275rr01.pdf", "url": "https://authors.library.caltech.edu/records/pxydn-wpc12/files/99275rr01.pdf" }, "resource_type": "article", "pub_year": "1999", "author_list": "Unger, M.; Kartalov, E.; et el." }, { "id": "https://authors.library.caltech.edu/records/q7kas-w4j63", "eprint_id": 85264, "eprint_status": "archive", "datestamp": "2023-08-19 04:48:14", "lastmod": "2023-10-18 18:01:51", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "England-P-M", "name": { "family": "England", "given": "Pamela M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Mapping Disulfide Connectivity Using Backbone Ester Hydrolysis", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1999 American Chemical Society. \n\nReceived June 22, 1999; Revised Manuscript Received August 20, 1999; Publication Date (Web): October 5, 1999. \n\nThis work was supported by grants from the National Institutes of Health (NS 34407 and NS 11756).", "abstract": "The site-specific incorporation of \u03b1-hydroxy acids into proteins using nonsense suppression can provide a powerful probe of protein structure and function. The resulting backbone ester may be selectively hydrolyzed in the presence of the peptide backbone, providing an \"orthogonal\" chemistry that can be useful both as an analytical tool and as a structural probe. Here we describe in detail a substantial substituent effect on this hydrolysis reaction. Consistent with mechanistic expectations, the steric bulk of the amino acid immediately N-terminal of the hydroxy acid has a large effect on the hydrolysis rate. On the basis of these results, we also describe a simple protocol for identifying disulfide loops in soluble and membrane proteins, exemplified by the \u03b1 subunit of the muscle nicotinic acetylcholine receptor (nAChR). If a backbone ester is incorporated outside a disulfide loop, hydrolysis alone gives two fragments, but if the ester is incorporated within a disulfide loop, both hydrolysis and reduction are required for cleavage. This test could be useful in characterizing the disulfide topology of complex, membrane proteins.", "date": "1999-10-26", "date_type": "published", "publication": "Biochemistry", "volume": "38", "number": "43", "publisher": "American Chemical Society", "pagerange": "14409-14415", "id_number": "CaltechAUTHORS:20180313-080855191", "issn": "0006-2960", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180313-080855191", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS 34407" }, { "agency": "NIH", "grant_number": "NS 11756" } ] }, "doi": "10.1021/bi991424c", "resource_type": "article", "pub_year": "1999", "author_list": "England, Pamela M.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/pj9p8-4fc15", "eprint_id": 99490, "eprint_status": "archive", "datestamp": "2023-08-22 14:01:28", "lastmod": "2023-10-18 18:20:53", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Khakh-B-S", "name": { "family": "Khakh", "given": "Baljit S." } }, { "id": "Proctor-W-R", "name": { "family": "Proctor", "given": "William R." } }, { "id": "Dunwiddie-T-V", "name": { "family": "Dunwiddie", "given": "Thomas V." } }, { "id": "Labarca-C-G", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Allosteric Control of Gating and Kinetics at P2X\u2084 Receptor Channels", "ispublished": "pub", "full_text_status": "public", "keywords": "ATP; ivermectin; ion channel; allosteric; modulation; P2X; purinoceptor", "note": "\u00a9 1999 Society for Neuroscience. \n\nReceived April 5, 1999; revised June 1, 1999; accepted June 14, 1999. \n\nThis work was supported by a Wellcome Trust (UK) International Prize Fellowship (to B.S.K.), National Institutes of Health Grant NS-11756 and NS-29173, and grants from the Department of Veterans Affairs. Thanks to Dr. I Chessell (Glaxo Wellcome) for providing P2X cDNA clones and Dr. Phillipe Seguela for the P2X6 cDNA. We thank S. McKinney and H. Li for help with preparation of hippocampal cultures and Xenopus oocytes and W. B. Smith for guidance on use of the Biolistics gene gun.\n\nPublished - 7289.full.pdf
", "abstract": "The CNS abundantly expresses P2X receptor channels for ATP; of these the most widespread in the brain is the P2X\u2084 channel. We show that ivermectin (IVM) is a specific positive allosteric effector of heterologously expressed P2X\u2084 and possibly of heteromeric P2X\u2084/P2X\u2086channels, but not of P2X\u2082, P2X\u2083, P2X\u2082/P2X\u2083, or P2X\u2087 channels. In the submicromolar range (EC\u2085\u2080, \u223c250 nM) the action of IVM was rapid and reversible, resulting in increased amplitude and slowed deactivation of P2X\u2084 channel currents evoked by ATP. IVM also markedly increased the potency of ATP and that of the normally low-potency agonist \u03b1,\u03b2-methylene-ATP in a use- and voltage-independent manner without changing the ion selectivity of P2X\u2084 channels. Therefore, IVM evokes a potent pharmacological gain-of-function phenotype that is specific for P2X\u2084 channels. We also tested whether IVM could modulate endogenously expressed P2X channels in the adult trigeminal mesencephalic nucleus and hippocampal CA1 neurons. Surprisingly, IVM produced no significant effect on the fast ATP-evoked inward currents in either type of neuron, despite the fact that IVM modulated P2X\u2084 channels heterologously expressed in embryonic hippocampal neurons. These results suggest that homomeric P2X\u2084 channels are not the primary subtype of P2X receptor in the adult trigeminal mesencephalic nucleus and in hippocampal CA1 neurons.", "date": "1999-09-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "19", "number": "17", "publisher": "Society for Neuroscience", "pagerange": "7289-7299", "id_number": "CaltechAUTHORS:20191028-115507054", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191028-115507054", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-29173" }, { "agency": "Department of Veterans Affairs" } ] }, "doi": "10.1523/jneurosci.19-17-07289.1999", "pmcid": "PMC6782529", "primary_object": { "basename": "7289.full.pdf", "url": "https://authors.library.caltech.edu/records/pj9p8-4fc15/files/7289.full.pdf" }, "resource_type": "article", "pub_year": "1999", "author_list": "Khakh, Baljit S.; Proctor, William R.; et el." }, { "id": "https://authors.library.caltech.edu/records/e0sg6-bm536", "eprint_id": 104084, "eprint_status": "archive", "datestamp": "2023-08-19 04:35:49", "lastmod": "2023-10-20 19:04:06", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Khakh-B-S", "name": { "family": "Khakh", "given": "Baljit S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Dynamic Selectivity Filters in Ion Channels", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1999 Cell Press. Published by Elsevier Under an Elsevier user license. \n\nThis work was supported by a Wellcome Trust (UK) International Prize Travelling Fellowship and the NIH (NS-11756).", "abstract": "Membrane ion channels contain integral pores that precisely select their permeant ions. This selectivity anchors our concepts of transmembrane signaling in many tissues, including the nervous system. Excitable cells have a rich repertoire of dynamically regulated channels, and this richness and plasticity allows them to use channels beautifully to suit their needs. The selectivity of ion channel pores has generally been viewed as fixed. However, recent studies on disparate classes of ion channels challenge the generality of this idea and show that some ion channels can change their ion selectivity such that normally impermeant ions do in fact permeate under some circumstances. In no case is the mechanism fully understood, but the phenomenon represents both a new functional aspect of ion channels and a suggestion about novel ways in which channels may process information in the nervous system. This review seeks to highlight studies on ion channels that show selectivity changes, point to possible mechanisms, and draw on common themes. We consider P2X and proton-gated channels from the superfamily of transmitter-gated ion channels, Kv and cardiac sodium channels from the superfamily of voltage-gated ion channels, and cyclic nucleotide\u2013gated channels from the family of channels that are gated by intracellular messengers (Figure 1).", "date": "1999-08", "date_type": "published", "publication": "Neuron", "volume": "23", "number": "4", "publisher": "Cell Press", "pagerange": "653-658", "id_number": "CaltechAUTHORS:20200626-123754307", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200626-123754307", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Wellcome Trust" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1016/s0896-6273(01)80025-8", "resource_type": "article", "pub_year": "1999", "author_list": "Khakh, Baljit S. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/62emf-czv93", "eprint_id": 104081, "eprint_status": "archive", "datestamp": "2023-08-19 04:20:25", "lastmod": "2023-10-20 19:03:44", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Jun", "name": { "family": "Li", "given": "Jun" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Functional roles of aromatic residues in the ligand-binding domain of cyclic nucleotide-gated channels", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1999 by the American Society for Pharmacology and Experimental Therapeutics. \n\nReceived June 2, 1998; accepted December 30, 1998. \n\nThis research was supported by Grant NS11756 from the National Institutes of Health. \n\nWe thank J. Ho for participating in the early phase of this project. We thank Dr. V. Kumar for the coordinates of the Brcng structural model and Dr. Y. Su for sharing the coordinates of PKA regulatory subunit before publication. We thank Drs. S. Scott and J. Tanaka for exchanging ideas. Dr. William Zagotta provided many stimulating discussions throughout this project.", "abstract": "The ligand-binding domains of cyclic nucleotide-gated (CNG) channels show sequence homology to corresponding region(s) of the Escherichia coli catabolite gene-activator protein (CAP) and to the regulatory subunit of cAMP-dependent or cGMP-dependent protein kinases. The structure of CAP and that of a cAMP-dependent protein kinases regulatory subunit have been solved, prompting efforts to generate structural models for the binding domains in CNG channel. These models explicitly predicted that an aromatic residue in the CNG channel aligning with leucine 61 of CAP forms an interaction with the bound cyclic nucleotide. We tested this hypothesis by site-directed mutagenesis in a rat olfactory channel (rOCNC1) and a bovine rod photoreceptor channel (Brcng). We found that mutations at this site had only weak effects that were not specific to the aromatic or the hydrophobic nature of the substituted residue. This result weakens the hypothesis of a strong or specific interaction at this site. We also separately mutated most of the other aromatic residues in the binding domain to alanine; most of these mutations resulted in channels that either did not function or had only minor changes in sensitivity. However, replacing tyrosine 565 with alanine (Y565A) in rOCNC1 increased agonist sensitivity by approximately 10-fold and resulted in prominent spontaneous activities. Y565 presumably lies between two alpha helices in the binding domain; one of these, the C helix, probably rotates during channel activation. The position of Y565 at the \"hinge\" between the C helix and another portion of the binding domain, and the consequences of Y565 mutations, strongly suggest that this portion of the binding domain is involved in channel gating processes.", "date": "1999-05", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "55", "number": "5", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "873-882", "id_number": "CaltechAUTHORS:20200626-123655847", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200626-123655847", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" } ] }, "resource_type": "article", "pub_year": "1999", "author_list": "Li, Jun and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/hstda-1eq78", "eprint_id": 104082, "eprint_status": "archive", "datestamp": "2023-08-19 04:20:32", "lastmod": "2023-10-20 19:03:49", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Jun", "name": { "family": "Li", "given": "Jun" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Single-channel kinetics of the rat olfactory cyclic nucleotide-gated channel expressed in Xenopus oocytes", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1999 by the American Society for Pharmacology and Experimental Therapeutics. \n\nReceived September 24, 1998; accepted January 28, 1999. \n\nThis research was supported by a grant from the National Institutes of Health (NS-11756). \n\nWe thank Yinong Zhang for much technical assistance and helpful discussions, Hairong Li and Brad Henkle for preparing oocytes, and Ben Edelman (Harvard College) for help using the MIL programs. We thank Dr. William Zagotta for insightful discussions during this project.", "abstract": "Cyclic nucleotide-gated channels are nonselective cation channels activated by intracellular cAMP and/or cGMP. It is not known how the binding of agonists opens the channel, or how the presumed four binding sites, one on each subunit, interact to generate cooperativity. We expressed the rat olfactory cyclic nucleotide-gated channel alpha subunit in Xenopus oocytes and recorded the single-channel currents. The channel had a single conductance state, and flickers at -60 mV showed the same power spectrum for cAMP and cGMP. At steady state, the distribution patterns of open and closed times were relatively simple, containing one or two exponential components. The conductance properties and the dwell-time distributions were adequately described by models that invoke only one or two binding events to open the channel, followed by an additional binding event that prolongs the openings and helps to explain apparent cooperativity. In a comparison between cAMP and cGMP, we find that cGMP has clearly higher binding affinity than cAMP, but only modestly higher probability of inducing the conformational transition that opens the channel.", "date": "1999-05", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "55", "number": "5", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "883-893", "id_number": "CaltechAUTHORS:20200626-123656741", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200626-123656741", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" } ] }, "resource_type": "article", "pub_year": "1999", "author_list": "Li, Jun and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/q04da-5j298", "eprint_id": 56626, "eprint_status": "archive", "datestamp": "2023-08-19 04:11:40", "lastmod": "2023-10-23 15:38:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Khakh-B-S", "name": { "family": "Khakh", "given": "Baljit S." } }, { "id": "Bao-Xiaoyan-R", "name": { "family": "Bao", "given": "Xiaoyan R." } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Neuronal P2X transmitter-gated cation channels change their ion selectivity in seconds", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1999 Nature America Inc.\n\nReceived 30 November 1998, Accepted 20 January 1999.\n\nThanks to I. Chessell (GlaxoWellcome, UK) for providing P2X cDNA clones. The authors are grateful to H. Li, S. McKinney and J. Sydes for assistance with preparation of oocytes and neurons and for advice on the YO-PRO-1 experiments, and to other members of the group for comments. This work was supported by the National Institutes of Health (NS-11756), a Wellcome Trust (UK) Prize Travelling Fellowship to B.S.K. and a Caltech Summer Undergraduate Research Fellowship to X.R.B.", "abstract": "Fast synaptic transmission depends on the selective ionic permeability of transmitter-gated ion channels. Here we show changes in the ion selectivity of neuronal P2X transmitter-gated cation channels as a function of time (on the order of seconds) and previous ATP exposure. Heterologously expressed P2X2, P2X2/P2X3 and P2X4 channels as well as native neuronal P2X channels possess various combinations of mono- or biphasic responses and permeability changes, measured by NMDG+ and fluorescent dye. Furthermore, in P2X4 receptors, this ability to alter ion selectivity can be increased or decreased by altering an amino-acid residue thought to line the ion permeation pathway, identifying a region that governs this activity-dependent change.", "date": "1999-04", "date_type": "published", "publication": "Nature Neuroscience", "volume": "2", "number": "4", "publisher": "Nature Publishing Group", "pagerange": "322-330", "id_number": "CaltechAUTHORS:20150414-095506624", "issn": "1097-6256", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150414-095506624", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Wellcome Trust" }, { "agency": "Caltech Summer Undergraduate Research Fellowship (SURF)" } ] }, "doi": "10.1038/7233", "resource_type": "article", "pub_year": "1999", "author_list": "Khakh, Baljit S.; Bao, Xiaoyan R.; et el." }, { "id": "https://authors.library.caltech.edu/records/cy6xr-n0g25", "eprint_id": 103214, "eprint_status": "archive", "datestamp": "2023-08-19 03:56:20", "lastmod": "2023-10-20 15:47:13", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "England-P-M", "name": { "family": "England", "given": "Pamela M." } }, { "id": "Zhang-Yinong", "name": { "family": "Zhang", "given": "Yinong" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Backbone Mutations in Transmembrane Domains of a Ligand-Gated Ion Channel: Implications for the Mechanism of Gating", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1999 Cell Press. Under an Elsevier user license. \n\nWe thank Cesar Labarca for gathering some of the data on conventional mutants in the M1 domain; Hong Dang for sharing unpublished information concerning the \u03b1 7 receptor; Sonia Pollit and Peter Schultz for samples of leucic acid and pipecolic acid; and members of our laboratory for much discussion. This research was supported by grants from the NIH (grants NS11756 and NS34407) and by National Research Service Awards to P. M. E. and Y. Z.", "abstract": "An approach to identify backbone conformational changes underlying nicotinic acetylcholine receptor (nAChR) gating was developed. Specific backbone peptide bonds were replaced with an ester, which disrupts backbone hydrogen bonds at the site of mutation. At a conserved proline residue (\u03b1Pro221) in the first transmembrane (M1) domain, the amide-to-ester mutation provides receptors with near-normal sensitivity, although the natural amino acids tested other than Pro produce receptors that gate with a much larger EC\u2085\u2080 than normal. Therefore, a backbone hydrogen bond at this site may interfere with normal gating. In the \u03b1M2 domain, the amide-to-ester mutation yielded functional receptors at 15 positions, 3 of which provided receptors with >10-fold lower EC\u2085\u2080 than wild type. These results support a model for gating that includes significant changes of backbone conformation within the M2 domain.", "date": "1999-01-08", "date_type": "published", "publication": "Cell", "volume": "96", "number": "1", "publisher": "Elsevier", "pagerange": "89-98", "id_number": "CaltechAUTHORS:20200514-152725398", "issn": "0092-8674", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200514-152725398", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1016/s0092-8674(00)80962-9", "resource_type": "article", "pub_year": "1999", "author_list": "England, Pamela M.; Zhang, Yinong; et el." }, { "id": "https://authors.library.caltech.edu/records/c9hg4-xq390", "eprint_id": 12574, "eprint_status": "archive", "datestamp": "2023-08-22 13:21:45", "lastmod": "2023-10-17 19:17:09", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "McPhee-J-C", "name": { "family": "McPhee", "given": "Jancy C." } }, { "id": "Dang-Yan-L", "name": { "family": "Dang", "given": "Yan L." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Evidence for a Functional Interaction between Integrins and G Protein-activated Inward Rectifier K+ Channels", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1998 by The American Society for Biochemistry and Molecular Biology, Inc. \n\n(Received for publication, August 17, 1998, and in revised form, September 29, 1998) \n\nWe thank M. Jasek for help with CHO cells; T. Ivanina for help with the Western blots and the manuscript; C. Doupnik, P. Kofuji, A. R. Horwitz, and D. Desimone for discussions; and R. Hynes and F. Lesage for antibodies. \n\nThis work was supported by National Institutes of Health Grant GM-29836. The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.\n\nPublished - MCPjbc98.pdf
", "abstract": "Heteromultimeric G protein-activated inward rectifier K+ (GIRK) channels, abundant in heart and brain, help to determine the cellular membrane potential as well as the frequency and duration of electrical impulses. The sequence arginine-glycine-aspartate (RGD), located extracellularly between the first membrane-spanning region and the pore, is conserved among all identified GIRK subunits but is not found in the extracellular domain of any other cloned K+ channels. Many integrins, which, like channels, are integral membrane proteins, recognize this RGD sequence on other proteins, usually in the extracellular matrix. We therefore asked whether GIRK activity might be regulated by direct interaction with integrin. Here, we present evidence that mutation of the RGD site to RGE, particularly on the GIRK4 subunit, decreases or abolishes GIRK current. Furthermore, wild-type channels can be co-immunoprecipitated with integrin. The total cellular amount of expressed mutant GIRK channel protein is the same as the wild-type protein; however, the amount of mutant channel protein that localizes to the plasma membrane is decreased relative to wild-type, most likely accounting for the diminished GIRK current detected. GIRK channels appear to bind directly to integrin and to require this interaction for proper GIRK channel membrane localization and function.", "date": "1998-12-25", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "273", "number": "52", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "34696-34702", "id_number": "CaltechAUTHORS:MCPjbc98", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:MCPjbc98", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-29836" } ] }, "primary_object": { "basename": "MCPjbc98.pdf", "url": "https://authors.library.caltech.edu/records/c9hg4-xq390/files/MCPjbc98.pdf" }, "resource_type": "article", "pub_year": "1998", "author_list": "McPhee, Jancy C.; Dang, Yan L.; et el." }, { "id": "https://authors.library.caltech.edu/records/6zkjs-cc024", "eprint_id": 113550, "eprint_status": "archive", "datestamp": "2023-08-22 13:21:03", "lastmod": "2023-10-23 17:46:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Yong-Xin", "name": { "family": "Li", "given": "Yong-Xin" } }, { "id": "Zhang-Yinong", "name": { "family": "Zhang", "given": "Yinong" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Schuman-E-M", "name": { "family": "Schuman", "given": "Erin M." }, "orcid": "0000-0002-7053-1005" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Enhancement of Neurotransmitter Release Induced by Brain-Derived Neurotrophic Factor in Cultured Hippocampal Neurons", "ispublished": "pub", "full_text_status": "public", "keywords": "BDNF; calcium; glutamate receptors; hippocampal; neurotrophins; synaptic plasticity; neurotransmitter release; General Neuroscience", "note": "\u00a9 1998 Society for Neuroscience. \n\nReceived July 29, 1998; revised Sept. 14, 1998; accepted Sept. 17, 1998. \n\nThis work was supported by the Silvio Conte Center for Neuroscience Research at Caltech sponsored by the National Institute of Mental Health and by Amgen, Thousand Oaks, CA.\n\nPublished - 10231.full.pdf
", "abstract": "Brain-derived neurotrophic factor (BDNF), like other neurotrophins, has long-term effects on neuronal survival and differentiation; furthermore, recent work has shown that BDNF also can induce rapid changes in synaptic efficacy. We have investigated the mechanism(s) of these synaptic effects on cultured embryonic hippocampal neurons. In the presence of the GABAA receptor antagonist, picrotoxin, the application of BDNF (100 ng/ml) for 1\u20135 min increased the amplitude of evoked synaptic currents by 48 \u00b1 9% in 10 of 15 pairs of neurons and increased the frequency of EPSC bursts to 205 \u00b1 20% of the control levels. There was no detectable effect of BDNF on various measures of electrical excitability, including the resting membrane potential, input resistance, action potential threshold, and action potential amplitude. In addition, BDNF did not change the postsynaptic currents induced by the exogenous application of glutamate. BDNF did increase the frequency of miniature EPSCs (mEPSCs) (268.0 \u00b1 46.8% of control frequency), however, without affecting the mEPSC amplitude. The effect of BDNF on mEPSC frequency was blocked by the tyrosine kinase inhibitor K252a and also by the removal of extracellular calcium ([Ca\u00b2\u207a]\u2092). Fura-2 recordings showed that BDNF elicited an increase in intracellular calcium concentration ([Ca\u00b2\u207a]\ua700). This effect was dependent on [Ca\u00b2\u207a]\u2092; it was blocked by K252a and by thapsigargin, but not by caffeine. The results demonstrate that BDNF enhances glutamatergic synaptic transmission at a presynaptic locus and that this effect is accompanied by a rise in [Ca\u00b2\u207a]\ua700 that requires the release of Ca\u00b2\u207a from IP3-gated stores.", "date": "1998-12-15", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "18", "number": "24", "publisher": "Society for Neuroscience", "pagerange": "10231-10240", "id_number": "CaltechAUTHORS:20220223-204480000", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220223-204480000", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Silvio Conte Center for Neuroscience Research" }, { "agency": "Amgen" } ] }, "doi": "10.1523/jneurosci.18-24-10231.1998", "pmcid": "PMC6793341", "primary_object": { "basename": "10231.full.pdf", "url": "https://authors.library.caltech.edu/records/6zkjs-cc024/files/10231.full.pdf" }, "resource_type": "article", "pub_year": "1998", "author_list": "Li, Yong-Xin; Zhang, Yinong; et el." }, { "id": "https://authors.library.caltech.edu/records/9sm9j-xpy98", "eprint_id": 903, "eprint_status": "archive", "datestamp": "2023-08-22 13:12:50", "lastmod": "2023-10-13 22:02:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Zhong-Wenge", "name": { "family": "Zhong", "given": "Wenge" } }, { "id": "Gallivan-J-P", "name": { "family": "Gallivan", "given": "Justin P." } }, { "id": "Zhang-Yinong", "name": { "family": "Zhang", "given": "Yinong" } }, { "id": "Li-Lintong", "name": { "family": "Li", "given": "Lintong" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "From ab initio quantum mechanics to molecular neurobiology: A cation-pi binding site in the nicotinic receptor", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotinic acetylcholine receptor, cation-pi interaction, unnatural amino acids, acetylcholine-receptor, amino-acids, directed mutagenesis, M2 domain, residues, recognition, resolution, aromatics, mutations, agonists", "note": "\u00a9 1998 by the National Academy of Sciences. \n\nCommunicated by John Abelson, California Institute of Technology, Pasadena, CA, August 6, 1998 (received for review June 3, 1998). \n\nWe thank M. W. Nowak, A. P. West, Jr., C. Labarca, H. Dang, Y. Tong, and Y. Li for helpful discussions. This work was supported by the National Institutes of Health (NS-34407 and NS-11756) and the California Tobacco-Related Disease Research Program. J.P.G. is grateful to the California Institute of Technology and to Eastman Kodak for fellowship support. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - ZHPpnas98.pdf
", "abstract": "The nicotinic acetylcholine receptor is the prototype ligand-gated ion channel. A number of aromatic amino acids have been identified as contributing to the agonist binding site, suggesting that cation-pi interactions may be involved in binding the quaternary ammonium group of the agonist, acetylcholine. Here we show a compelling correlation between: (i) ab initio quantum mechanical predictions of cation-pi binding abilities and (ii) EC50 values for acetylcholine at the receptor for a series of tryptophan derivatives that were incorporated into the receptor by using the in vivo nonsense-suppression method for unnatural amino acid incorporation. Such a correlation is seen at one, and only one, of the aromatic residues-tryptophan-149 of the alpha subunit. This finding indicates that, on binding, the cationic, quaternary ammonium group of acetylcholine makes van der Waals contact with the indole side chain of alpha tryptophan-149, providing the most precise structural information to date on this receptor. Consistent with this model, a tethered quaternary ammonium group emanating from position alpha 149 produces a constitutively active receptor.", "date": "1998-10-13", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "95", "number": "21", "publisher": "National Academy of Sciences", "pagerange": "12088-12093", "id_number": "CaltechAUTHORS:ZHPpnas98", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:ZHPpnas98", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Caltech" }, { "agency": "Eastman Kodak Company" } ] }, "doi": "10.1073/pnas.95.21.12088", "pmcid": "PMC22789", "primary_object": { "basename": "ZHPpnas98.pdf", "url": "https://authors.library.caltech.edu/records/9sm9j-xpy98/files/ZHPpnas98.pdf" }, "resource_type": "article", "pub_year": "1998", "author_list": "Zhong, Wenge; Gallivan, Justin P.; et el." }, { "id": "https://authors.library.caltech.edu/records/78r7f-95d37", "eprint_id": 113552, "eprint_status": "archive", "datestamp": "2023-08-22 13:12:16", "lastmod": "2023-10-23 17:46:27", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Cao-Yongwei", "name": { "family": "Cao", "given": "Yongwei" } }, { "id": "Li-Ming", "name": { "family": "Li", "given": "Ming" } }, { "id": "Mager-Sela", "name": { "family": "Mager", "given": "Sela" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Amino Acid Residues that Control pH Modulation of Transport-Associated Current in Mammalian Serotonin Transporters", "ispublished": "pub", "full_text_status": "public", "keywords": "5-HT; Xenopus oocyte; protons; channels; electrophysiology; sodium; General Neuroscience", "note": "\u00a9 1998 Society for Neuroscience. \n\nReceived June 23, 1998; accepted July 14, 1998. \n\nThis work was supported by Grant DA-09121 from the National Institute on Drug Abuse and Grant NS-11756 from the National Institute of Neurological Disorders and Stroke, by a National Institutes of Health National Research Service Award to Y.C., and by a National Alliance for Research on Schizophrenia and Depression fellowship to S.M. We thank B. J. Hoffman and G. Rudnick for providing cDNAs.\n\nPublished - 7739.full.pdf
", "abstract": "The rat and human serotonin transporters (rSERT and hSERT, respectively) were expressed in Xenopus oocytes and studied using site-directed mutagenesis, electrophysiological recordings, and [\u00b3H]5-HT uptake measurements. rSERT, but not hSERT, displayed increased transport-associated current at low pH. Chimeras and point mutations showed that, of the 52 nonidentical residues, a single residue at position 490 (threonine in rSERT and lysine in hSERT) governs this difference. Furthermore, potentiation required the glutamate residue at position 493. Cysteine substitution and alkylation experiments showed that residue 493 is extracellular. Cysteine at 493 increased, whereas aspartate decreased, the net charge movement per transported 5-HT molecule. The mutations at this region did not significantly affect other aspects of SERT function, including agonist-independent leakage current, voltage-dependent transient current, and H\u207a current. This region may therefore be part of an external gate required for rSERT function. The data and analyses show that, in the absence of detailed structural information, a gate\u2013lumen\u2013gate scheme is useful for interpreting results from mutations that alter functional properties of neurotransmitter transporters.", "date": "1998-10-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "18", "number": "19", "publisher": "Society for Neuroscience", "pagerange": "7739-7749", "id_number": "CaltechAUTHORS:20220223-204499000", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220223-204499000", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "National Alliance for Research on Schizophrenia and Depression" } ] }, "doi": "10.1523/jneurosci.18-19-07739.1998", "pmcid": "PMC6793029", "primary_object": { "basename": "7739.full.pdf", "url": "https://authors.library.caltech.edu/records/78r7f-95d37/files/7739.full.pdf" }, "resource_type": "article", "pub_year": "1998", "author_list": "Cao, Yongwei; Li, Ming; et el." }, { "id": "https://authors.library.caltech.edu/records/fgk10-rz292", "eprint_id": 86271, "eprint_status": "archive", "datestamp": "2023-08-22 13:08:35", "lastmod": "2023-10-18 19:25:20", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "The Crystal Structure of a Potassium Channel \u2014 A New Era in the Chemistry of Biological Signaling", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Ion channels; Membrane proteins; Protein structures; Signal transduction; Structure elucidation", "note": "\u00a9 1998 Wiley\u2010VCH Verlag GmbH, Weinheim, Fed. Rep. of Germany. \n\nIssue Online 17 December 1998; Version of Record online: 17 December 1998. \n\nWe thank Professors Doug Rees (California Institute of Technology), Chris Miller (Brandeis), and Rod MacKinnon (Rockefeller) for helpful discussions. Our own work on K^+ channels and related structures is supported by the NIH (NS-34407 and GM 29836).", "abstract": "The similarity to crown ethers is apparent when the arrangement of the oxygen atoms of the carbonyl groups of the protein backbone in the structure of the potassium channel (see schematic drawing of a section of the structure) found in the bacterium Streptomyces lividans is considered. This particular part of the channel pore acts as the selectivity filter, with the permeability of the channel for K+ being as much as 10\u2009000 times greater than for the Na+ ion. In fact, in this area of the structure two K+ ions are located, a feature that enables high flux through the channel.", "date": "1998-09-18", "date_type": "published", "publication": "Angewandte Chemie International Edition", "volume": "37", "number": "17", "publisher": "Wiley", "pagerange": "2329-2331", "id_number": "CaltechAUTHORS:20180508-083019710", "issn": "1433-7851", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180508-083019710", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "GM 29836" } ] }, "doi": "10.1002/(SICI)1521-3773(19980918)37:17<2329::AID-ANIE2329>3.0.CO;2-M", "resource_type": "article", "pub_year": "1998", "author_list": "Dougherty, Dennis A. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/0e4a0-ejg41", "eprint_id": 905, "eprint_status": "archive", "datestamp": "2023-08-22 13:06:38", "lastmod": "2023-10-23 17:48:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Yong-Xin", "name": { "family": "Li", "given": "Yong-Xin" } }, { "id": "Xu-Youfeng", "name": { "family": "Xu", "given": "Youfeng" } }, { "id": "Ju-Donghong", "name": { "family": "Ju", "given": "Donghong" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Schuman-E-M", "name": { "family": "Schuman", "given": "Erin M." }, "orcid": "0000-0002-7053-1005" } ] }, "title": "Expression of a dominant negative TrkB receptor, T1, reveals a requirement for presynaptic signaling in BDNF-induced synaptic potentiation in cultured hippocampal neurons", "ispublished": "pub", "full_text_status": "public", "keywords": "LONG-TERM POTENTIATION, NEUROTROPHIC FACTOR, SILENT SYNAPSES, TYROSINE KINASE, PLASTICITY, LTP, TRANSMISSION, ENHANCEMENT, MICE", "note": "\u00a9 1998 National Academy of Sciences. \n\nContributed by Norman Davidson, July 10, 1998. \n\nWe thank Dr. Andy Welcher (Amgen) for the generous gift of BDNF and TrkB antibody. We thank Sheri McKinney for tissue culture work. This work was supported by National Institutes of Health Grant MH-49176. E.M.S. is a Pew Biomedical Scholar, Beckman Young Investigator, Merck Scholar, and an Assistant Investigator of the Howard Hughes Medical Institute. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - LIYpnas98.pdf
", "abstract": "We have developed a method to analyze the relative contributions of pre- and postsynaptic actions of a particular gene product in neurons in culture and potentially in slices using adenovirus-mediated gene transfer. A recombinant virus directed the expression of both a GFP reporter protein and TrkB.T1, a C-terminal truncated dominant negative TrkB neurotrophin receptor. When expressed in the presynaptic cell at synapses between embryonic hippocampal neurons in culture, the dominant negative TrkB.T1 inhibited two forms of synaptic potentiation induced by the neurotrophin brain-derived neurotrophic factor (BDNF): (i) greater evoked synaptic transmission and (ii) higher frequency of spontaneous miniature synaptic currents. These inhibition effects are not seen if the transgene is expressed only in the postsynaptic cell. We conclude that BDNF-TrkB signal transduction in the presynaptic terminal leads to both types of potentiation and is therefore the primary cause of synaptic enhancement by BDNF in these neurons.", "date": "1998-09-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "95", "number": "18", "publisher": "National Academy of Sciences", "pagerange": "10884-10889", "id_number": "CaltechAUTHORS:LIYpnas98", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LIYpnas98", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "Pew Charitable Trust" }, { "agency": "Merck" }, { "agency": "Howard Hughes Medical Institute (HHMI)" } ] }, "pmcid": "PMC27990", "primary_object": { "basename": "LIYpnas98.pdf", "url": "https://authors.library.caltech.edu/records/0e4a0-ejg41/files/LIYpnas98.pdf" }, "resource_type": "article", "pub_year": "1998", "author_list": "Li, Yong-Xin; Xu, Youfeng; et el." }, { "id": "https://authors.library.caltech.edu/records/jjpdt-fw429", "eprint_id": 103176, "eprint_status": "archive", "datestamp": "2023-08-19 03:18:23", "lastmod": "2023-10-20 15:44:14", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Silverman-S-K", "name": { "family": "Silverman", "given": "Scott K." }, "orcid": "0000-0001-8166-3460" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Asymmetrical Contributions of Subunit Pore Regions to Ion Selectivity in an Inward Rectifier K\u207a Channel", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1998 The Biophysical Society. Published by Elsevier Inc. \n\nReceived 18 February 1998, Accepted 2 June 1998.\n\nWe thank Drs. Paulo Kofuji and Craig Doupnik for discussions. \n\nThis work was supported by grants from the National Institutes of Health (NS34407 and GM29836).", "abstract": "We have investigated aspects of ion selectivity in K\u207a channels by functional expression of wild-type and mutant heteromultimeric G protein-coupled inward-rectifier K\u207a (GIRK) channels in Xenopus oocytes. Within the K\u207a channel pore (P) region signature sequence, a large number of point mutations in GIRK1 and GIRK4 subunits have been made at a key tyrosine residue\u2014the \"signature\" tyrosine of the GYG. Studies of mutant GIRK1/GIRK4 heteromultimers reveal that the GIRK1 and GIRK4 subunits contribute asymmetrically to K\u207a selectivity. The signature tyrosine of GIRK1 can be mutated to many different residues while retaining selectivity; in contrast, the analogous position in GIRK4 must be tyrosine for maximum selectivity. Other residues of the P region also contribute to selectivity, and studies with GIRK1/GIRK4 chimeras reveal that an intact, heteromultimeric P region is necessary and sufficient for optimal K\u207a selectivity. We propose that the GIRK1 and GIRK4 P regions play roles similar to the two P regions of an emerging family of K\u207a channels whose subunits each have two P regions connected in tandem. We find different consequences between similar mutations in inward-rectifier and voltage-gated K\u207a channels, which suggests that the pore structures and selectivity mechanisms in the two classes of channel may not be identical. We confirm that GIRK4 subunits alone can form functional channels in oocytes, but we find that these channels are measurably permeable to Na\u207a and Ca\u00b2\u207a.", "date": "1998-09", "date_type": "published", "publication": "Biophysical Journal", "volume": "75", "number": "3", "publisher": "Biophysical Society", "pagerange": "1330-1339", "id_number": "CaltechAUTHORS:20200513-104033054", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200513-104033054", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "GM29836" } ] }, "doi": "10.1016/s0006-3495(98)74051-2", "pmcid": "PMC1299807", "resource_type": "article", "pub_year": "1998", "author_list": "Silverman, Scott K.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/gna9v-7hj90", "eprint_id": 103239, "eprint_status": "archive", "datestamp": "2023-08-19 02:41:59", "lastmod": "2023-10-20 15:49:16", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Miller-Jeffrey-C", "name": { "family": "Miller", "given": "Jeffrey C." } }, { "id": "Silverman-S-K", "name": { "family": "Silverman", "given": "Scott K." } }, { "id": "England-P-M", "name": { "family": "England", "given": "Pamela M." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Flash Decaging of Tyrosine Sidechains in an Ion Channel", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1998 Cell Press. Published by Elsevier Inc. \n\nReceived 17 December 1997, Revised 9 February 1998. \n\nThis work was supported by grants from the National Institutes of Health (NS-11756, NS-34407). We thank Mark W. Nowak, Patrick C. Kearney, and Rory Sayres for help and Justin Gallivan for continuing advice. J. C. M. held Caltech undergraduate research fellowships; P. M. E. held an NRSA.\n\nSupplemental Material - 1-s2.0-S0896627300810016-mmc1.gif
", "abstract": "A nonsense codon suppression technique was employed to incorporate ortho-nitrobenzyl tyrosine, \"caged tyrosine,\" in place of tyrosine at any of three positions (93, 127, or 198) in the \u03b1 subunit of the muscle nicotinic ACh receptor (nAChR) expressed in Xenopus oocytes. The ortho-nitrobenzyl group was then removed by 1 ms flashes at 300\u2013350 nm to yield tyrosine itself while macroscopic currents were recorded during steady ACh exposure. Responses to multiple flashes showed (1) that each flash decages up to 17% of the tyrosines and (2) that two tyrosines must be decaged per receptor for a response. The conductance relaxations showed multiple kinetic components; rate constants (<0.1 s\u207b\u00b9 to 10\u00b3 s\u207b\u00b9) depended on pH and the site of incorporation, and relative amplitudes depended on the number of prior flashes. This method, which is potentially quite general, (1) provides a time-resolved assay for the behavior of a protein when a mutant sidechain is abruptly changed to the wild-type residue and (2) will also allow for selective decaging of sidechains that are candidates for covalent modification (such as phosphorylation) in specific proteins in intact cells.", "date": "1998-04", "date_type": "published", "publication": "Neuron", "volume": "20", "number": "4", "publisher": "Cell Press", "pagerange": "619-624", "id_number": "CaltechAUTHORS:20200515-132054818", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200515-132054818", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "Caltech" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1016/s0896-6273(00)81001-6", "primary_object": { "basename": "1-s2.0-S0896627300810016-mmc1.gif", "url": "https://authors.library.caltech.edu/records/gna9v-7hj90/files/1-s2.0-S0896627300810016-mmc1.gif" }, "resource_type": "article", "pub_year": "1998", "author_list": "Miller, Jeffrey C.; Silverman, Scott K.; et el." }, { "id": "https://authors.library.caltech.edu/records/svfdx-a6t17", "eprint_id": 6585, "eprint_status": "archive", "datestamp": "2023-08-22 12:36:10", "lastmod": "2023-10-16 20:25:04", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "New Views of Multi-Ion Channels", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1998 by The Rockefeller University Press. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. \n\nCommentary.\n\nPublished - LESjgp98.pdf
", "abstract": "Thus, most site-directed mutagenesis data render it untenable to consider that two or more roughly equivalent high affinity sites govern selectivity in multi-ion pores. The papers by Dang and McCleskey and Kiss et al. respond to this challenge by showing that a model with a single high affinity site, flanked by two binding sites of lower affinity close to the pore entrances, can generate much of the classical multi-ion behavior. The sites need not interact, and the two flanking sites could arise from one of several mechanisms: a featureless charged vestibule, a dehydration step, or a specific weak binding site. \n\nThe multi-ion pore remains a cornerstone of permeation theory, but the new theory features only a single high affinity site and no mutual repulsion. The high flux rate occurs because ions pause at the flanking sites and reequilibrate thermally, gaining enough energy to move over the next barrier.", "date": "1998-02", "date_type": "published", "publication": "Journal of General Physiology", "volume": "111", "number": "2", "publisher": "Rockefeller University Press", "pagerange": "181-183", "id_number": "CaltechAUTHORS:LESjgp98", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LESjgp98", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1085/jgp.111.2.181", "pmcid": "PMC2222761", "primary_object": { "basename": "LESjgp98.pdf", "url": "https://authors.library.caltech.edu/records/svfdx-a6t17/files/LESjgp98.pdf" }, "resource_type": "article", "pub_year": "1998", "author_list": "Lester, Henry A. and Dougherty, Dennis A." }, { "id": "https://authors.library.caltech.edu/records/fpveh-4z566", "eprint_id": 6139, "eprint_status": "archive", "datestamp": "2023-08-22 11:57:33", "lastmod": "2023-10-16 20:09:28", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "West-A-P-Jr", "name": { "family": "West", "given": "Anthony P., Jr." } }, { "id": "Bjorkman-P-J", "name": { "family": "Bjorkman", "given": "Pamela J." }, "orcid": "0000-0002-2277-3990" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Expression and Circular Dichroism Studies of the Extracellular Domain of the alpha Subunit of the Nicotinic Acetylcholine Receptor", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1997 The American Society for Biochemistry and Molecular Biology, Inc. \n\n(Received for publication, April 21, 1997, and in revised form, July 23, 1997) \n\nWe thank David Penny for help with the Cell Pharm expression, Shelley Diamond for assistance with flow cytometry, Steve Mayo for assistance with the CD experiments, and Jon Lindstrom for providing mAb 210. \n\nThis work was supported by National Institutes of Health Grants NS-11756 and NS-34407, the California Tobacco-related Disease Research Program, and the Howard Hughes Medical Institute.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.\n\nPublished - WESjbc97.pdf
", "abstract": "To provide material suitable for structural studies of the nicotinic acetylcholine receptor, we have expressed and purified the NH2-terminal extracellular domain of the mouse muscle alpha subunit. Several constructs were initially investigated using Xenopus oocytes as a convenient small scale expression system. A fusion protein (alpha210GPI) consisting of the 210 NH2-terminal amino acids of the alpha subunit and a glycosylphosphatidylinositol anchorage sequence conferred surface alpha-bungarotoxin binding in oocytes. Coexpression of alpha210GPI with an analogous construct made from the delta subunit showed no evidence of heterodimer formation. The alpha210GPI protein was chosen for large scale expression in transfected Chinese hamster ovary cells. The alpha210GPI protein was cleaved from these cells and purified on an immunoaffinity column. Gel and column chromatography show that the purified protein is processed as expected and exists as a monomer. The purified protein also retains the two distinct, conformation-specific binding sites expected for the correctly folded alpha subunit. Circular dichroism studies of alpha210GPI suggest that this region of the receptor includes considerable beta-sheet secondary structure, with a small proportion of alpha-helix.", "date": "1997-10-10", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "272", "number": "41", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "25468-25473", "id_number": "CaltechAUTHORS:WESjbc97", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:WESjbc97", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Howard Hughes Medical Institute (HHMI)" } ] }, "doi": "10.1074/jbc.272.41.25468", "primary_object": { "basename": "WESjbc97.pdf", "url": "https://authors.library.caltech.edu/records/fpveh-4z566/files/WESjbc97.pdf" }, "resource_type": "article", "pub_year": "1997", "author_list": "West, Anthony P., Jr.; Bjorkman, Pamela J.; et el." }, { "id": "https://authors.library.caltech.edu/records/nq7ec-c6d79", "eprint_id": 103246, "eprint_status": "archive", "datestamp": "2023-08-22 11:56:18", "lastmod": "2023-10-20 15:49:51", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Gallivan-J-P", "name": { "family": "Gallivan", "given": "Justin P." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Site-specific incorporation of biotinylated amino acids to identify surface-exposed residues in integral membrane proteins", "ispublished": "pub", "full_text_status": "restricted", "keywords": "biotin/streptavidin; in vivo suppression; surface exposure; transmembrane topology; unnatural amino acids", "note": "\u00a9 1997 Published by Elsevier Ltd. \n\nReceived 24 July 1997, Accepted 27 August 1997. \n\nThis work was supported by the National Institutes of Health (NS-34407 and NS-11756). J.P.G. is grateful to Caltech and to the Eastman Kodak Corporation for support in the form of graduate fellowships. We thank Scott K. Silverman for providing a sample of \u03b1-NVOC-lysine fluorenylmethyl ester.", "abstract": "Background: A key structural issue for all integral membrane proteins is the exposure of individual residues to the intracellular or extracellular media. This issue involves the basic transmembrane topology as well as more subtle variations in surface accessibility. Direct methods to evaluate the degree of exposure for residues in functional proteins expressed in living cells would be highly valuable. We sought to develop a new experimental method to determine highly surface-exposed residues, and thus transmembrane topology, of membrane proteins expressed in Xenopus oocytes. \n\nResults: We have used the in vivo nonsense suppression technique to incorporate biotinylated unnatural amino acids into functional ion channels expressed in Xenopus oocytes. Binding of \u00b9\u00b2\u2075I-streptavidin to biotinylated receptors was used to determine the surface exposure of individual amino acids. In particular, we studied the main immunogenic region of the nicotinic acetylcholine receptor. The biotin-containing amino acid biocytin was efficiently incorporated into five sites in the main immunogenic region and extracellular streptavidin bound to one residue in particular, \u03b170. The position of \u03b170 as highly exposed on the receptor surface was thus established. \n\nConclusions: The in vivo nonsense suppression technique has been extended to provide the first in a potential series of methods to identify exposed residues and to assess their relative exposure in functional proteins expressed in Xenopus oocytes.", "date": "1997-10", "date_type": "published", "publication": "Chemistry and Biology", "volume": "4", "number": "10", "publisher": "Elsevier", "pagerange": "739-749", "id_number": "CaltechAUTHORS:20200515-142830762", "issn": "1074-5521", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200515-142830762", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Caltech" }, { "agency": "Eastman Kodak Corporation" } ] }, "doi": "10.1016/s1074-5521(97)90312-4", "resource_type": "article", "pub_year": "1997", "author_list": "Gallivan, Justin P.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/030t7-f8b69", "eprint_id": 906, "eprint_status": "archive", "datestamp": "2023-08-22 11:55:14", "lastmod": "2023-10-13 22:02:15", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "England-P-M", "name": { "family": "England", "given": "Pamela M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Site-specific, photochemical proteolysis applied to ion channels in vivo", "ispublished": "pub", "full_text_status": "public", "keywords": "nicotinic acetylcholine receptor, (2-nitrophenyl)glycine, K+ channel, signature disulfide loop, site-specific, nitrobenzyl-induced photochemical proteolysis, conserved disulfide loop, unnatural amino-acids, potassium channel, binding-site, chemical aminoacylation, photolabile precursors, caged compounds, alpha-subunit, M2 domain", "note": "\u00a9 1997 by the National Academy of Sciences. \n\nCommunicated by Peter B. Dervan, California Institute of Technology, Pasadena, CA, July 11, 1997 (received for review April 20, 1997). \n\nWe thank P. Reinhart, P. Kearney, M. Nowak, and S. Silverman for suggestions and helpful discussions and B. Henkle and H. Li for assistance with oocytes. This work was supported by the National Institutes of Health (NS-34407, NS-11756, and a National Research Service Award to P.M.E.) and the University of California Tobacco-Related Disease Research Program. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - ENGpnas97.pdf
", "abstract": "A method for site-specific, nitrobenzyl-induced photochemical proteolysis of diverse proteins expressed in living cells has been developed based on the chemistry of the unnatural amino acid (2-nitrophenyl)glycine (Npg). Using the in vivo nonsense codon suppression method for incorporating unnatural amino acids into proteins expressed in Xenopus oocytes, Npg has been incorporated into two ion channels: the Drosophila Shaker B K+ channel and the nicotinic acetylcholine receptor. Functional studies in vivo show that irradiation of proteins containing an Npg residue does lead to peptide backbone cleavage at the site of the novel residue. Using this method, evidence is obtained for an essential functional role of the \"signature\" Cys128-Cys142 disulfide loop of the nAChR alpha subunit.", "date": "1997-09-30", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "94", "number": "20", "publisher": "National Academy of Sciences", "pagerange": "11025-11030", "id_number": "CaltechAUTHORS:ENGpnas97", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:ENGpnas97", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "California Tobacco-Related Disease Research Program" } ] }, "doi": "10.1073/pnas.94.20.11025", "pmcid": "PMC23572", "primary_object": { "basename": "ENGpnas97.pdf", "url": "https://authors.library.caltech.edu/records/030t7-f8b69/files/ENGpnas97.pdf" }, "resource_type": "article", "pub_year": "1997", "author_list": "England, Pamela M.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/g2669-h3v47", "eprint_id": 915, "eprint_status": "archive", "datestamp": "2023-08-22 11:54:13", "lastmod": "2023-10-23 19:38:25", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Doupnik-C-A", "name": { "family": "Doupnik", "given": "Craig A." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Kofuji-Paulo", "name": { "family": "Kofuji", "given": "Paulo" } } ] }, "title": "RGS proteins reconstitute the rapid gating kinetics of G\u03b2\u03b3-activated inwardly rectifying K^+\u2009channels", "ispublished": "pub", "full_text_status": "public", "keywords": "POTASSIUM CHANNEL, ALPHA-SUBUNITS, I-KACH, RECEPTOR, INHIBITION, FAMILY, CELLS, GENE, DESENSITIZATION, IDENTIFICATION", "note": "\u00a9 1997 by the National Academy of Sciences. \n\nContributed by Norman Davidson, July 17, 1997. \n\nWe thank Drs. Kirk Druey and John Kehrl for providing RGS1\u20133 cDNAs, Brad Henkle and Hai-Rong Li for preparing oocytes, and Dr. Mark Jasek for helping with the mammalian cell transfection procedures. This work was supported by fellowships from the American Heart Association (C.A.D. and P.K.) and the Guenther Foundation (P.K.), and by grants from the National Institutes of Health and the National Institute of Mental Health. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - DOUpnas97.pdf
", "abstract": "G protein gated inward rectifier K+ (GIRK) channels mediate hyperpolarizing postsynaptic potentials in the nervous system and in the heart during activation of G alpha(i/o) coupled receptors. In neurons and cardiac atrial cells the time course for receptor-mediated GIRK current deactivation is 20-40 times faster than that observed in heterologous systems expressing cloned receptors and GIRK channels, suggesting that an additional component(s) is required to confer the rapid kinetic properties of the native transduction pathway. We report here that heterologous expression of \"regulators of G protein signaling\" (RGS proteins), along with cloned G protein-coupled receptors and GIRK channels, reconstitutes the temporal properties of the native receptor --> GIRK signal transduction pathway. GIRK current waveforms evoked by agonist activation of muscarinic m(2) receptors or serotonin 1A receptors were dramatically accelerated by coexpression of either RGS1, RGS3, or RGS4, but not RGS2. For the brain-expressed RGS4 isoform, neither the current amplitude nor the steady-state agonist dose-response relationship was significantly affected by RGS expression, although the agonist-independent \"basal\" GIRK current was suppressed by approximate to 40%. Because GIRK activation and deactivation kinetics are the limiting rates for the onset and termination of \"slow\" postsynaptic inhibitory currents in neurons and atrial cells, RGS proteins may play crucial roles in the timing of information transfer within the brain and to peripheral tissues.", "date": "1997-09-16", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "94", "number": "19", "publisher": "National Academy of Sciences", "pagerange": "10461-10466", "id_number": "CaltechAUTHORS:DOUpnas97", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:DOUpnas97", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "American Heart Association" }, { "agency": "Guenther Foundation" }, { "agency": "NIH" }, { "agency": "National Institute of Mental Health (NIMH)" } ] }, "pmcid": "PMC23385", "primary_object": { "basename": "DOUpnas97.pdf", "url": "https://authors.library.caltech.edu/records/g2669-h3v47/files/DOUpnas97.pdf" }, "resource_type": "article", "pub_year": "1997", "author_list": "Doupnik, Craig A.; Davidson, Norman; et el." }, { "id": "https://authors.library.caltech.edu/records/vj1en-bjk20", "eprint_id": 918, "eprint_status": "archive", "datestamp": "2023-08-22 11:46:21", "lastmod": "2023-10-23 19:41:23", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Ehrengruber-M-U", "name": { "family": "Ehrengruber", "given": "Markus U." } }, { "id": "Doupnik-C-A", "name": { "family": "Doupnik", "given": "Craig A." } }, { "id": "Xu-Youfeng", "name": { "family": "Xu", "given": "Youfeng" } }, { "id": "Garvey-J-S", "name": { "family": "Garvey", "given": "Justine" } }, { "id": "Jasek-M-C", "name": { "family": "Jasek", "given": "Mark C." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Activation of heteromeric G protein-gated inward rectifier K+ channels overexpressed by adenovirus gene transfer inhibits the excitability of hippocampal neurons", "ispublished": "pub", "full_text_status": "public", "keywords": "functional expression, mediated transfer, rat hippocampus, pyramidal cells, receptors, serotonin, cloning, family, electrophysiology, rectification", "note": "\u00a9 1997 National Academy of Sciences. \n\nContributed by Norman Davidson, April 16, 1997. \n\nWe thank S. L. McKinney for the primary cell cultures, B. W. Henkle for oocyte preparation, C. Chavkin for help with GIRK antibodies, P. Kofuji for construction of the Shaker plasmid and helpful comments, C. Lin for construction of Ad5HT1AR, S. J. Stary for advice on the adenovirus technique, and L. Byerly for instruction in electrophysiology. This work was supported by the National Institute of Mental Health, National Institute of General Medical Sciences, Human Frontier Science Program, the Swiss National Science Foundation (Fellowships 81BE-40054 and 823A-042966 to M.U.E.), and the American Heart Association (fellowship to C.A.D.). \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - EHRpnas97.pdf
", "abstract": "G protein-gated inward rectifier K+ channel subunits 1-4(GIRK1-4) have been cloned from neuronal and atrial tissue and function as heterotetramers. To examine the inhibition of neuronal excitation by GIRKs, we overexpressed GIRKs in cultured hippocampal neurons from 18 day rat embryos, which normally lack or show low amounts of GIRK protein and currents. Adenoviral recombinants containing the cDNAs for GIRK1, GIRK2, GIRK4, and the serotonin 1A receptor were constructed. Typical GIRK currents could be activated by endogenous GABA(B), serotonin 5-HT1A, and adenosine A1 receptors in neurons coinfected with GIRK1+2 or GIRK1+4. Under current clamp, GIRK activation increased the cell membrane conductance by 1- to 2-fold, hyperpolarized the cell by 11-14 mV,and inhibited action potential firing by increasing, the threshold current for firing by 2- to 3-fold. These effects were not found in non- and mock-infected neurons, and were similar to the effects of muscarinic stim ulation of native GIRK currents in atrial myocytes. Two inhibitory effects of GIRK activation, hyperpolarization and diminution of depolarizing pulses, were simulated from the experimental data. These inhibitory effects are physiologically important in the voltage range between the resting membrane potential and the potential where voltage-gated Na+ and K+ currents are activated; that is where GIRK currents are outward.", "date": "1997-06-24", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "94", "number": "13", "publisher": "National Academy of Sciences", "pagerange": "7070-7075", "id_number": "CaltechAUTHORS:EHRpnas97", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:EHRpnas97", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Institute of Mental Health (NIMH)" }, { "agency": "NIH" }, { "agency": "National Institute of General Medical Sciences" }, { "agency": "Human Frontier Science Program" }, { "agency": "Swiss National Science Foundation (SNSF)", "grant_number": "81BE-40054" }, { "agency": "Swiss National Science Foundation (SNSF)", "grant_number": "823A-042966" }, { "agency": "American Heart Association" } ] }, "pmcid": "PMC21286", "primary_object": { "basename": "EHRpnas97.pdf", "url": "https://authors.library.caltech.edu/records/vj1en-bjk20/files/EHRpnas97.pdf" }, "resource_type": "article", "pub_year": "1997", "author_list": "Ehrengruber, Markus U.; Doupnik, Craig A.; et el." }, { "id": "https://authors.library.caltech.edu/records/7r36m-a2f93", "eprint_id": 99563, "eprint_status": "archive", "datestamp": "2023-08-22 11:41:30", "lastmod": "2023-10-18 18:33:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Quick-M-W", "name": { "family": "Quick", "given": "Michael W." } }, { "id": "Corey-J-L", "name": { "family": "Corey", "given": "Janis L." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Second Messengers, Trafficking-Related Proteins, and Amino Acid Residues that Contribute to the Functional Regulation of the Rat Brain GABA Transporter GAT1", "ispublished": "pub", "full_text_status": "public", "keywords": "neurotransmitter transporters; intracellular trafficking; leucine heptad repeats; synaptic vesicle proteins; second\nmessengers; protein regulation", "note": "\u00a9 1997 Society for Neuroscience. \n\nReceived Aug. 9, 1996; revised Jan. 23, 1997; accepted Feb. 13, 1997. \n\nThis research was supported by United States Public Health Service Grants NS-11756 (H.A.L.), DA-09121 (H.A.L.), DA-10509 (M.W.Q.), National Research Service Award fellowships (M.W.Q. and J.L.C.), and the W. M. Keck Foundation 931360 (M.W.Q).\n\nPublished - 2967.full.pdf
", "abstract": "Recent evidence indicates that several members of the Na\u207a-coupled transporter family are regulated, and this regulation in part occurs by redistribution of transporters between intracellular locations and the plasma membrane. We elucidate components of this process for both wild-type and mutant GABA transporters (GAT1) expressed in Xenopus oocytes using a combination of uptake assays, immunoblots, and electrophysiological measurements of membrane capacitance, transport-associated currents, and GAT1-specific charge movements. At low GAT1 expression levels, activators of protein kinase C (PKC) induce redistribution of GAT1 from intracellular vesicles to the plasma membrane; at higher GAT1 expression levels, activators of PKC fail to induce this redistribution. However, coinjection of total rat brain mRNA with GAT1 permits PKC-mediated modulation at high transporter expression levels. This effect of brain mRNA on modulation is mimicked by coinjection of syntaxin 1a mRNA and is eliminated by injecting synaptophysin or syntaxin antisense oligonucleotides. Additionally, botulinum toxins, which inactivate proteins involved in vesicle release and recycling, reduce basal GAT1 expression and prevent PKC-induced translocation. Mutant GAT1 proteins, in which most or all of a leucine heptad repeat sequence was removed, display altered basal distribution and lack susceptibility to modulation by PKC, delineating one region of GAT1 necessary for its targeting. Thus, functional regulation of GAT1 in oocytes occurs via components common to transporters and to trafficking in both neural and non-neural cells, and suggests a relationship between factors that control neurotransmitter secretion and the components necessary for neurotransmitter uptake.", "date": "1997-05-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "17", "number": "9", "publisher": "Society for Neuroscience", "pagerange": "2967-2979", "id_number": "CaltechAUTHORS:20191030-140944005", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191030-140944005", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "DA-10509" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "W. M. Keck Foundation", "grant_number": "931360" } ] }, "doi": "10.1523/jneurosci.17-09-02967.1997", "pmcid": "PMC6573650", "primary_object": { "basename": "2967.full.pdf", "url": "https://authors.library.caltech.edu/records/7r36m-a2f93/files/2967.full.pdf" }, "resource_type": "article", "pub_year": "1997", "author_list": "Quick, Michael W.; Corey, Janis L.; et el." }, { "id": "https://authors.library.caltech.edu/records/9de29-s1152", "eprint_id": 4317, "eprint_status": "archive", "datestamp": "2023-08-22 11:39:27", "lastmod": "2023-10-16 17:42:30", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Li-Jun", "name": { "family": "Li", "given": "Jun" } }, { "id": "Zagotta-W-N", "name": { "family": "Zagotta", "given": "William N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Cyclic nucleotide-gated channels: structural basis of ligand efficacy and allosteric modulation", "ispublished": "pub", "full_text_status": "public", "keywords": "GMP-SENSITIVE CONDUCTANCE; DEPENDENT PROTEIN-KINASE; ROD OUTER SEGMENTS; RETINAL RODS; ACTIVATED CHANNELS; CATION CHANNEL; BINDING DOMAIN; ION CHANNELS; SUBUNIT INTERACTIONS; MOLECULAR MECHANISM", "note": "\u00a9 1997 Cambridge University Press. Reprinted with permission. \n\nThe authors thank members of the Zagotta laboratory for stimulating discussions. Sela Mager, Mark Nowak, Yinong Zhang and Jing Liu read the manuscript critically. The preparation of this review is supported by research grants from the National Institute of Health: EY-10329, MH-49176, GM-29836, and by predoctoral training grant GM-08501. W.N.Z. is an Investigator of the Howard Hughes Medical Institute.\n\nPublished - LIJqrb97.pdf
", "abstract": "Most working proteins, including metabolic enzymes, transcription regulators, and membrane receptors, transporters, and ion channels, share the property of allosteric coupling. The term 'allosteric' means that these proteins mediate indirect interactions between sites that are physically separated on the protein. In the example of ligand-gated ion channels, the binding of a suitable ligand elicits local conformational changes at the binding site, which are coupled to further conformational changes in regions distant from the binding site. The physical motions finally arrive at the site of biological activity: the ion-permeating pore. The conformational changes that lead from the ligand binding to the actual opening of the pore comprise 'gating'. In 1956, del Castillo and Katz suggested that the competition between different ligands at nicotinic acetylcholine receptors (nAChRs) could be explained by formation of an intermediate, ligand-bound, yet inactive state of the receptor, which separates the active state of the receptor from the initial binding of the ligand (del Castillo & Katz, 1957). This 'binding-then-gating', two-step model went beyond the then-prevailing drug-receptor model that assumes a single bimolecular binding reaction, and paralleled Stephenson's conceptual dichotomy of 'affinity' and 'efficacy' (Stephenson, 1956). In 1965 Monod, Wyman and Changeux presented a simple allosteric model (the MWC model) (Monod et al. 1965) that explained the cooperative binding of oxygen to haemoglobin; it was adopted as an important paradigm for ligand-gated channels soon after its initial formulation (Changeux et al. 1967; Karlin, 1967; Colquhoun, 1973).", "date": "1997-05", "date_type": "published", "publication": "Quarterly Reviews of Biophysics", "volume": "30", "number": "2", "publisher": "Cambridge University Press", "pagerange": "177-193", "id_number": "CaltechAUTHORS:LIJqrb97", "issn": "0033-5835", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LIJqrb97", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "EY-10329" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM-08501" }, { "agency": "Howard Hughes Medical Institute (HHMI)" } ] }, "primary_object": { "basename": "LIJqrb97.pdf", "url": "https://authors.library.caltech.edu/records/9de29-s1152/files/LIJqrb97.pdf" }, "resource_type": "article", "pub_year": "1997", "author_list": "Li, Jun; Zagotta, William N.; et el." }, { "id": "https://authors.library.caltech.edu/records/0b7pp-9ts82", "eprint_id": 99564, "eprint_status": "archive", "datestamp": "2023-08-19 01:14:34", "lastmod": "2023-10-18 18:33:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Cao-Yongwei", "name": { "family": "Cao", "given": "Yongwei" } }, { "id": "Mager-S", "name": { "family": "Mager", "given": "Sela" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "H\u207a Permeation and pH Regulation at a Mammalian Serotonin Transporter", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1997 Society for Neuroscience. \n\nReceived Oct. 15, 1996; revised Jan. 3, 1997; accepted Jan. 7, 1997. \n\nThis work was supported by grants from the National Institute on Drug Abuse (DA-09121) and the National Institute of Neurological Diseases and Stroke (NS-11756) and by a National Institutes of Health National Research Service Award to Y.C. We thank F. Lin for participating in some of the experiments, M. Sonders for discussing his data with us, and N. Davidson for comments.\n\nPublished - 2257.full.pdf
", "abstract": "The rat serotonin transporter expressed in Xenopusoocytes displays an inward current in the absence of 5-HT when external pH is lowered to 6.5 or below. The new current differs from the leakage current described previously in two ways. (1) It is \u223c10-fold larger at pH 5 than the leakage current at pH 7.5 and reaches 1000 H\u207a/sec per transporter at extremes of voltage and pH with no signs of saturation. (2) It is selective for H\u207a by reversal potential measurements. Similar H\u207a-induced currents are also observed in several other ion-coupled transporters, including the GABA transporter, the dopamine transporter, and the Na\u207a/glucose transporter. The high conductance and high selectivity of the H\u207a-induced current suggest that protons may be conducted via a hydrogen-bonded chain (a \"proton-wire mechanism\") formed at least partially by side chains within the transporter. In addition, pH affects other conducting states of rat serotonin transporter. Acidic pH potentiates the 5-HT-induced, transport-associated current and inhibits the hyperpolarization-activated transient current. The dose\u2013response relationships for these two effects suggest that two H\u207a binding sites, with pK_a values close to 5.1 and close to 6.3, govern the potentiation of the 5-HT-induced current and the inhibition of the transient current, respectively. These results are important for developing structure-function models that explain permeation properties of neurotransmitter transporters.", "date": "1997-04-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "17", "number": "7", "publisher": "Society for Neuroscience", "pagerange": "2257-2266", "id_number": "CaltechAUTHORS:20191030-144551486", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191030-144551486", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "DA-09121" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "National Institute on Drug Abuse" }, { "agency": "National Institute of Neurological Disorders and Stroke (NINDS)" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1523/jneurosci.17-07-02257.1997", "pmcid": "PMC6573487", "primary_object": { "basename": "2257.full.pdf", "url": "https://authors.library.caltech.edu/records/0b7pp-9ts82/files/2257.full.pdf" }, "resource_type": "article", "pub_year": "1997", "author_list": "Cao, Yongwei; Mager, Sela; et el." }, { "id": "https://authors.library.caltech.edu/records/25ssr-zb989", "eprint_id": 65857, "eprint_status": "archive", "datestamp": "2023-08-19 01:09:21", "lastmod": "2023-10-18 16:52:15", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Bradley-J", "name": { "family": "Bradley", "given": "Jonathan" } }, { "id": "Zhang-Yinong", "name": { "family": "Zhang", "given": "Yinong" } }, { "id": "Bakin-R", "name": { "family": "Bakin", "given": "Robert" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Ronnett-G-V", "name": { "family": "Ronnett", "given": "Gabriele V." } }, { "id": "Zinn-K", "name": { "family": "Zinn", "given": "Kai" }, "orcid": "0000-0002-6706-5605" } ] }, "title": "Functional Expression of the Heteromeric \"Olfactory\" Cyclic Nucleotide-Gated Channel in the Hippocampus: A Potential Effector of Synaptic Plasticity in Brain Neurons", "ispublished": "pub", "full_text_status": "public", "keywords": "cyclic nucleotide-gated channels cAMP cGMP olfaction sensory transduction hippocampus synaptic plasticity", "note": "\u00a9 1997 Society for Neuroscience. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). \n\nReceived Nov. 11, 1996; revised Dec. 23, 1996; accepted Dec. 30, 1996. \n\nThis work was supported by a grant from the National Institutes of Mental Health to K.Z., and by grants from the National Institute of Deafness and Communication Disorders and the W. M. Keck Foundation to G.R. J.B. was supported by a National Institutes of Health (NIH) graduate training grant and Awards for Research College Scientists Foundation. Y.Z. was supported by a National Research Service Award postdoctoral fellowship from NIH. We thank Norman Davidson, Mary Kennedy, Jun Li, Ming-Ji Fann, Chris Schoenherr, Yasuhito Uezono, David Anderson, Cori Bargmann, and members of the Zinn, Ronnett, Lester, Davidson, Anderson, and Kennedy groups for helpful discussions; Susan Ou and the Caltech Monoclonal Antibody Facility for generation of hybridomas; and Sheri McKinney for hippocampal cultures.\n\nPublished - 1993.full.pdf
", "abstract": "Cyclic nucleotide-gated (cng) channels are important components of signaling systems mediating sensory transduction. In vertebrate photoreceptors, light activates a signaling cascade that causes a decrease in intracellular cGMP concentrations, closing retinal cng channels. Signal transduction in olfactory receptor neurons is believed to proceed via G-protein-mediated elevation of intracellular cAMP in response to odorant binding by 7-helix receptors. cAMP opens the olfactory cng channel, which is highly permeable to Ca^(2+). Here we demonstrate by in situ hybridization and immunohistochemistry with subunit-specific antibodies that both subunits of the heteromeric rat olfactory cng channel are also widely expressed in the brain. Expression of the retinal rod cng channel, however, can be detected only in the eye. In the adult hippocampus, the olfactory cng channel is expressed on cell bodies and processes of CA1 and CA3 neurons. In cultured embryonic hippocampal neurons, the channel is localized to a subset of growth cones and processes. We recorded conductances with the electrophysiological characteristics of the heteromeric olfactory cng channel in excised inside-out patches from these cultured neurons. We also show that Ca^(2+) influx into hippocampal neurons in response to cyclic nucleotide elevation can be detected using fura-2 imaging. Cyclic nucleotide elevation has been implicated in several mechanisms of synaptic plasticity in the hippocampus, and these mechanisms also require elevation of intracellular Ca^(2+). Our results suggest that the \"olfactory\" cng channel could regulate synaptic efficacy in brain neurons by modulating Ca^(2+) levels in response to changes in cyclic nucleotide concentrations.", "date": "1997-03-15", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "17", "number": "6", "publisher": "Society for Neuroscience", "pagerange": "1993-2005", "id_number": "CaltechAUTHORS:20160401-135631881", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160401-135631881", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Institute of Mental Health (NIMH)" }, { "agency": "National Institute on Deafness and Other Communication Disorders" }, { "agency": "W. M. Keck Foundation" }, { "agency": "NIH" }, { "agency": "Research College Scientists Foundation" } ] }, "doi": "10.1523/JNEUROSCI.17-06-01993.1997", "pmcid": "PMC6793760", "primary_object": { "basename": "1993.full.pdf", "url": "https://authors.library.caltech.edu/records/25ssr-zb989/files/1993.full.pdf" }, "resource_type": "article", "pub_year": "1997", "author_list": "Bradley, Jonathan; Zhang, Yinong; et el." }, { "id": "https://authors.library.caltech.edu/records/36jff-scs18", "eprint_id": 914, "eprint_status": "archive", "datestamp": "2023-08-22 11:25:09", "lastmod": "2023-10-13 22:02:33", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Silverman-S-K", "name": { "family": "Silverman", "given": "Scott K." } }, { "id": "Kofuji-P", "name": { "family": "Kofuji", "given": "Paulo" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "A regenerative link in the ionic fluxes through the weaver potassium channel underlies the pathophysiology of the mutation", "ispublished": "pub", "full_text_status": "public", "keywords": "RECTIFYING K+ CHANNELS, XENOPUS-LAEVIS OOCYTES, MUTANT MOUSE, CALCIUM, CLONING, DIFFERENTIATION, EXPRESSION, BRAIN", "note": "\u00a9 1996 by the National Academy of Sciences. \n\nContributed by Norman Davidson, October 16, 1996. \n\nWe thank N. Dascal for comments. This work was supported by grants from the National Institute of Mental Health (MH-49176), the National Institute of General Medical Sciences (GM-29836), and the National Institute of Neurological Disorders and Stroke (NS-34407). P.K. held fellowships from the Guenther Foundation and the American Heart Association. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - SILpnas96.pdf
", "abstract": "The homozygous weaver mouse displays neuronal degeneration in several brain regions. Previous experiments in heterologous expression systems showed that the G protein-gated inward rectifier K+ channel (GIRK2) bearing the weaver pore-region GYG-to-SYG mutation (i) is not activated by G(beta gamma) subunits, but instead shows constitutive activation, and (ii) is no longer a K+-selective channel but conducts Na+ as well. The present experiments on weaverGIRK2 (wv-GIRK2) expressed in Xenopus oocytes show that the level of constitutive activation depends on Intracellular Na+ concentration. In particular, manipulations that decrease intracellular Na+ produce a component of Na+-permeable current activated via a G protein pathway. Therefore, constitutive activation may not arise because the weaver mutation directly alters the gating transitions of the channel protein. Instead, there may be a regenerative cycle of Na+ influx through the wvGIRK2 channel, leading to additional Na+ activation. We also show that the wvGIRK2 channel is permeable to Ca2+, providing an additional mechanism for the degeneration that characterizes the weaver phenotype. We further demonstrate that the GIRK4 channel bearing the analogous weaver mutation has properties similar to those of the wvGIRK2 channel, providing a glimpse of the selective pressures that have maintained the GYG sequence in nearly all known K+ channels.", "date": "1996-12-24", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "93", "number": "26", "publisher": "National Academy of Sciences", "pagerange": "15429-15434", "id_number": "CaltechAUTHORS:SILpnas96", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:SILpnas96", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "NS-34407" }, { "agency": "Guenther Foundation" }, { "agency": "American Heart Association" } ] }, "doi": "10.1073/pnas.93.26.15429", "pmcid": "PMC26421", "primary_object": { "basename": "SILpnas96.pdf", "url": "https://authors.library.caltech.edu/records/36jff-scs18/files/SILpnas96.pdf" }, "resource_type": "article", "pub_year": "1996", "author_list": "Silverman, Scott K.; Kofuji, Paulo; et el." }, { "id": "https://authors.library.caltech.edu/records/e6csj-gev35", "eprint_id": 4668, "eprint_status": "archive", "datestamp": "2023-08-22 11:24:27", "lastmod": "2023-10-16 17:52:21", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Quick-M-W", "name": { "family": "Quick", "given": "Michael W." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Simon-M-I", "name": { "family": "Simon", "given": "Melvin I." } }, { "id": "Aragay-A-M", "name": { "family": "Aragay", "given": "Anna M." } } ] }, "title": "Desensitization of Inositol 1,4,5-Trisphosphate/Ca2+-induced Cl- Currents by Prolonged Activation of G Proteins in Xenopus Oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1996 by The American Society for Biochemistry and Molecular Biology, Inc. \n\n(Received for publication, June 26, 1996, and in revised form, September 12, 1996) \n\nWe thank Brad Henkle for oocyte preparation and Lorna Brundage for critical reading of the manuscript. We are also grateful to Dr. T. Wilkie for providing the Galpha 15Q121L cDNA. \n\nThis work was supported in part by W. M. Keck Foundation Grant 931360 (to M. W. Q.) and National Institute of Health Grants CA13148 (to M. W. Q.) and MH-49176 (to M. I. S. and H. A. L.). The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.\n\nPublished - QUIjbc96.pdf
", "abstract": "Expression of G protein alpha subunits of the Gq family with various G protein-coupled receptors induces activation of an inositol 1,4,5-trisphosphate (IP3)/Ca2+-mediated Cl- conductance in Xenopus oocytes. Our present data show that two members of this family, the human Galpha 16 subunit and the murine homologue Galpha 15, can induce both activation and inhibition of these agonist-induced currents. Although extremely low amounts (10-50 pg) of injected Galpha 16 subunit cRNA cause modest (~2-fold) enhancement of ligand-induced Cl- currents in oocytes co-injected with thyrotropin-releasing hormone (TRH) receptor cRNA 48 h postinjection, larger Galpha 16 and Galpha 15 cRNA injections cause >10-fold inhibition of TRH or 5HT2c receptor responses. The inhibition is analyzed in this study. The inhibited currents are recovered if various Gbeta gamma subunit combinations are also expressed with the Galpha subunits. The constitutively active mutant, Galpha 16Q212L, also causes a strong attenuation of the ligand-induced Cl- currents, but this inhibition is not recovered by co-expression of Gbeta gamma subunits. These results indicate that the free Galpha subunit is responsible for the inhibitory signal. Although expression of TRH receptor alone produces maximum responses approximately 48 h after injection, co-expression of TRH receptor with Galpha 16 results in enhanced responses 6-12 h postinjection, followed by complete attenuation at 36 h. Furthermore, injection of Galpha 16 cRNA alone at comparable levels gives rise to spontaneous Cl- currents within 6-12 h postinjection, suggesting that the early spontaneous activation underlies the later suppression. Expression of other G protein alpha subunits of the Gq family, at cRNA levels considerably higher than effective for Galpha 16, produces both analogous spontaneous Cl- currents and, later, inhibition of ligand-induced Cl- currents. Experiments with direct injection of IP3 and of Ca2+ suggest that this inhibition is consistent with the down-regulation of IP3 receptors. These data indicate that both enhancement and inhibition of signaling through G protein-coupled receptors can be mediated by the expression level and/or activity of an individual G protein.", "date": "1996-12-13", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "271", "number": "50", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "32021-32027", "id_number": "CaltechAUTHORS:QUIjbc96", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:QUIjbc96", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "W. M. Keck Foundation", "grant_number": "931360" }, { "agency": "NIH", "grant_number": "CA13148" }, { "agency": "NIH", "grant_number": "MH-49176" } ] }, "primary_object": { "basename": "QUIjbc96.pdf", "url": "https://authors.library.caltech.edu/records/e6csj-gev35/files/QUIjbc96.pdf" }, "resource_type": "article", "pub_year": "1996", "author_list": "Quick, Michael W.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/2dx9z-pd019", "eprint_id": 103252, "eprint_status": "archive", "datestamp": "2023-08-19 00:39:46", "lastmod": "2023-10-20 15:50:12", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kearney-P-C", "name": { "family": "Kearney", "given": "Patrick C." } }, { "id": "Zhang-Haiyun", "name": { "family": "Zhang", "given": "Haiyun" } }, { "id": "Zhong-Wenge", "name": { "family": "Zhong", "given": "Wenge" } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Determinants of Nicotinic Receptor Gating in Natural and Unnatural Side Chain Structures at the M2 9\u2032 Position", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1996 Cell Press. Published by Elsevier Inc. \n\nReceived 13 September 1996, Revised 16 October 1996. \n\nWe thank Brad Henkle for help with oocytes and H. Dang, P. England, C. Labarca, M. Nowak, M. E. Saks, J. Sampson, S. Silverman, and Y. Zhang for comments. This work was supported by the National Institutes of Health (NS34407 and NS11756), by the California Tobacco-Related Disease Research Program, and by the Beckman Institute at Caltech. \n\nThe costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 USC Section 1734 solely to indicate this fact.", "abstract": "A nonsense suppression method was employed to incorporate a total of four natural and six unnatural residues at the 9\u2032 position of the M2 region in the \u03b2, \u03b3, and \u03b4 subunits of muscle nicotinic receptors. In 33 pairwise comparisons of functional properties as influenced by structural features including side chain length, branching, and substitution of oxygen for methylene carbons, it is concluded that increased polarity in the side chains at the 9\u2032 position consistently increases the sensitivity to acetylcholine. In addition, the stereochemistry of the side chain can have marked influences on the EC\u2085\u2080, primarily because of changes in the single-channel open time. For the case of isoleucine versus allo-isoleucine in the \u03b4 subunit, these changes are themselves modified by mutations at the 9\u2032 position in other subunits. The data suggest an especially strong interaction between the \u03b2 and \u03b4 subunits in the pore region, leading in turn to a suggested arrangement of subunits within the pentamer.", "date": "1996-12", "date_type": "published", "publication": "Neuron", "volume": "17", "number": "6", "publisher": "Cell Press", "pagerange": "1221-1229", "id_number": "CaltechAUTHORS:20200518-070216780", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200518-070216780", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Caltech Beckman Institute" } ] }, "doi": "10.1016/s0896-6273(00)80252-4", "resource_type": "article", "pub_year": "1996", "author_list": "Kearney, Patrick C.; Zhang, Haiyun; et el." }, { "id": "https://authors.library.caltech.edu/records/a8s9m-2tf70", "eprint_id": 29176, "eprint_status": "archive", "datestamp": "2023-08-19 00:36:07", "lastmod": "2023-10-24 18:25:07", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Silverman-S-K", "name": { "family": "Silverman", "given": "Scott K." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Subunit Stoichiometry of a Heteromultimeric G protein-coupled Inward-rectifier K^+ Channel", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1996 by The American Society for Biochemistry and Molecular Biology, Inc.\nReceived June 5, 1996. \nRevision received July 23, 1996. \n\nWe thank Paulo Kofuji for many helpful\ndiscussions.\nThis work was supported by National Institutes of Health Grants\nMH49176, GM29836, NS11756, and NS34407. The costs of publication\nof this article were defrayed in part by the payment of page charges.\nThis article must therefore be hereby marked \"advertisement\" in accordance\nwith 18 U.S.C. Section 1734 solely to indicate this fact.\n\nPublished - SILjbc96.pdf
", "abstract": "We investigated the stoichiometry of the heteromultimeric G protein-coupled inward-recitfier K^+ channel (GIRK) formed from GIRK1 and GIRK4 subunits. Multimeric GIRK constructs with several concatenated channel subunits were expressed in Xenopus oocytes. Coexpression of various trimeric constructs with different monomers clearly showed that the functional channel has stoichiometry (GIRK1)_2(GIRK4)_2. Efforts to establish a preferred arrangement of subunits around the channel pore suggest that more than one arrangement may be viable.", "date": "1996-11-29", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "271", "number": "48", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "30524-30528", "id_number": "CaltechAUTHORS:20120207-144820693", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120207-144820693", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "MH49176" }, { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" } ] }, "doi": "10.1074/jbc.271.48.30524", "primary_object": { "basename": "SILjbc96.pdf", "url": "https://authors.library.caltech.edu/records/a8s9m-2tf70/files/SILjbc96.pdf" }, "resource_type": "article", "pub_year": "1996", "author_list": "Silverman, Scott K.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/m5ngc-8cv63", "eprint_id": 104083, "eprint_status": "archive", "datestamp": "2023-08-19 00:32:28", "lastmod": "2023-10-20 19:03:57", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kearney-P-C", "name": { "family": "Kearney", "given": "Patrick C." } }, { "id": "Nowak-M-W", "name": { "family": "Nowak", "given": "Mark W." } }, { "id": "Zhong-Wenge", "name": { "family": "Zhong", "given": "Wenge" } }, { "id": "Silverman-S-K", "name": { "family": "Silverman", "given": "Scott K." }, "orcid": "0000-0001-8166-3460" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "Dose-response relations for unnatural amino acids at the agonist binding site of the nicotinic acetylcholine receptor: tests with novel side chains and with several agonists", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1996 by the American Society for Pharmacology and Experimental Therapeutics. \n\nReceived June 10, 1996; Accepted August 3, 1996. \n\nThis work was supported by the National Institutes of Health (Grants NS34407 and NS11756) and by the Beckman Institute at Caltech. \n\nWe thank Dr. Margaret E. Saks and Dr. Jeffrey R. Sampson for providing T7 polymerase and helpful advice.", "abstract": "Structure-function relations in the nicotinic acetylcholine receptor are probed using a recently developed method based on chemical synthesis of nonsense suppressor tRNAs with unnatural amino acid residues, site-directed incorporation at nonsense codons in Xenopus laevis oocytes, and electrophysiological measurements. A broad range of unnatural amino acids, as many as 14 at a given site, are incorporated at three sites, alpha 93, alpha 190, and alpha 198, all of which are tyrosine in the wild-type receptor and are thought to contribute to the agonist binding site. Confirming and expanding upon earlier studies using conventional mutagenesis, the three tyrosines are shown to be in substantially different structural microenvironments. In particular, a crucial role is established for the hydroxyl group of alpha Tyr93, whereas a variety of substituents are functional at the analogous position of alpha Tyr198. Interestingly, consideration of three different agonists (acetylcholine, nicotine, and tetramethylammonium) does not discriminate between these two best-characterized binding site residues. In addition, double-mutation studies establish the independent effects of mutations at the pore region (second transmembrane region) and at the agonist binding site, and this observation leads to a novel strategy for adjusting EC\u2085\u2080 values. These results establish the broad generality and great potential of the unnatural amino acid methodology for illuminating subtle structural distinctions in neuroreceptors and related integral membrane proteins.", "date": "1996-11", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "50", "number": "5", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "1401-1412", "id_number": "CaltechAUTHORS:20200626-123657699", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200626-123657699", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "Caltech Beckman Institute" } ] }, "resource_type": "article", "pub_year": "1996", "author_list": "Kearney, Patrick C.; Nowak, Mark W.; et el." }, { "id": "https://authors.library.caltech.edu/records/1crnh-hza75", "eprint_id": 4271, "eprint_status": "archive", "datestamp": "2023-08-22 11:14:48", "lastmod": "2023-10-16 17:41:11", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Saks-M-E", "name": { "family": "Saks", "given": "Margaret E." } }, { "id": "Sampson-J-R", "name": { "family": "Sampson", "given": "Jeffrey R." } }, { "id": "Nowak-M-W", "name": { "family": "Nowak", "given": "Mark W." } }, { "id": "Kearney-P-C", "name": { "family": "Kearney", "given": "Patrick C." } }, { "id": "Du-Fangyong", "name": { "family": "Du", "given": "Fangyong" } }, { "id": "Abelson-J-N", "name": { "family": "Abelson", "given": "John N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" } ] }, "title": "An engineered Tetrahymena tRNA(Gln) for in vivo incorporation of unnatural amino acids into proteins by nonsense suppression", "ispublished": "pub", "full_text_status": "public", "keywords": "TRANSFER RNA-SYNTHETASE; NICOTINIC ACETYLCHOLINE-RECEPTOR; GATED ION CHANNELS; M2 DOMAIN; RECOGNITION; ANTICODON; MUTATIONS; IDENTITY; GENES; ISOPENTENYLATION", "note": "\u00a9 1996 by The American Society for Biochemistry and Molecular Biology, Inc. \n\nReceived for publication, April 1, 1996, and in revised form, June 11, 1996. \n\nThis research was supported by National Institutes of Health Grants NS11756 (to H.A.L.), NS34407 (to D.A.D.), and GM48560 (to J.N.A.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.\n\nPublished - SAKjbc96.pdf
", "abstract": "A new tRNA, THG73, has been designed and evaluated as a vehicle for incorporating unnatural amino acids site-specifically into proteins expressed in vivo using the stop codon suppression technique. The construct is a modification of tRNAGln(CUA) from Tetrahymena thermophila, which naturally recognizes the stop codon UAG. Using electrophysiological studies of mutations at several sites of the nicotinic acetylcholine receptor, it is established that THG73 represents a major improvement over previous nonsense suppressors both in terms of efficiency and fidelity of unnatural amino acid incorporation. Compared with a previous tRNA used for in vivo suppression, THG73 is as much as 100-fold less likely to be acylated by endogenous synthetases of the Xenopus oocyte. This effectively eliminates a major concern of the in vivo suppression methodology, the undesirable incorporation of natural amino acids at the suppression site. In addition, THG73 is 4-10-fold more efficient at incorporating unnatural amino acids in the oocyte system. Taken together, these two advances should greatly expand the range of applicability of the in vivo nonsense suppression methodology.", "date": "1996-09-20", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "271", "number": "38", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "23169-23175", "id_number": "CaltechAUTHORS:SAKjbc96", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:SAKjbc96", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "NS34407" }, { "agency": "NIH", "grant_number": "GM48560" } ] }, "primary_object": { "basename": "SAKjbc96.pdf", "url": "https://authors.library.caltech.edu/records/1crnh-hza75/files/SAKjbc96.pdf" }, "resource_type": "article", "pub_year": "1996", "author_list": "Saks, Margaret E.; Sampson, Jeffrey R.; et el." }, { "id": "https://authors.library.caltech.edu/records/kqt24-rw065", "eprint_id": 99571, "eprint_status": "archive", "datestamp": "2023-08-22 11:13:25", "lastmod": "2023-10-18 18:33:59", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Mager-Sela", "name": { "family": "Mager", "given": "Sela" } }, { "id": "Kleinberger-Doron-Nurit", "name": { "family": "Kleinberger-Doron", "given": "Nurit" } }, { "id": "Keshet-Gilmor-I", "name": { "family": "Keshet", "given": "Gilmor I." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Kanner-Baruch-I", "name": { "family": "Kanner", "given": "Baruch I." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Ion Binding and Permeation at the GABA Transporter GAT1", "ispublished": "pub", "full_text_status": "public", "keywords": "sodium; chloride; GAT1; GABA; transporter; Xenopus oocyte", "note": "\u00a9 1996 Society for Neuroscience. \n\nReceived April 26, 1996; revised June 14, 1996; accepted June 18, 1996. \n\nThis work was supported by grants from the National Institute of Neurological Diseases and Stroke and the U.S./Israel Binational Science Foundation, and by fellowships from the Lester Deutsch Foundation and the Muscular Dystrophy Association (S.M.). We thank G. Rudnick for help with the [3H]tiagabine binding assay, C. Armstrong and A. Finkelstein for discussion of charge movements and membrane dielectrics, and J. Li and M. Nowak for comments on this manuscript.\n\nPublished - 5405.full.pdf
", "abstract": "This study addresses the binding of ions and the permeation of substrates during function of the GABA transporter GAT1. GAT1 was expressed in Xenopus oocytes and studied electrophysiologically as well as with [\u00b3H]GABA flux; GAT1 was also expressed in mammalian cells and studied with [\u00b3H]GABA and [\u00b3H]tiagabine binding. Voltage jumps, Na\u207a and Cl\u207b concentration jumps, and exposure to high-affinity blockers (NO-05-711 and SKF-100330A) all produce capacitive charge movements. Occlusive interactions among these three types of perturbations show that they all measure the same population of charges. The concentration dependences of the charge movements reveal (1) that two Na\u207a ions interact with the transporter even in the absence of GABA, and (2) that Cl\u207b facilitates the binding of Na\u207a. Comparison between the charge movements and the transport-associated current shows that this initial Na\u207a-transporter interaction limits the overall transport rate when [GABA] is saturating. However, two classes of manipulation\u2014treatment with high-affinity uptake blockers and the W68L mutation\u2014\"lock\" Na\u207a onto the transporter by slowing or preventing the subsequent events that release the substrates to the intracellular medium. The Na\u207a substitutes Li\u207a and Cs\u207a do not support charge movements, but they can permeate the transporter in an uncoupled manner. Our results (1) support the hypothesis that efficient removal of synaptic transmitter by the GABA transporter GAT1 depends on the previous binding of Na\u207a and Cl\u207b, and (2) indicate the important role of the conserved putative transmembrane domain 1 in interactions with the permeant substrates.", "date": "1996-09-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "16", "number": "17", "publisher": "Society for Neuroscience", "pagerange": "5405-5414", "id_number": "CaltechAUTHORS:20191030-155045434", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191030-155045434", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Institute of Neurological Disorders and Stroke (NINDS)" }, { "agency": "NIH" }, { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "Lester Deutsch Foundation" }, { "agency": "Muscular Dystrophy Association" } ] }, "doi": "10.1523/jneurosci.16-17-05405.1996", "pmcid": "PMC6578888", "primary_object": { "basename": "5405.full.pdf", "url": "https://authors.library.caltech.edu/records/kqt24-rw065/files/5405.full.pdf" }, "resource_type": "article", "pub_year": "1996", "author_list": "Mager, Sela; Kleinberger-Doron, Nurit; et el." }, { "id": "https://authors.library.caltech.edu/records/atfxh-7fk74", "eprint_id": 58161, "eprint_status": "archive", "datestamp": "2023-08-20 07:29:40", "lastmod": "2023-10-23 19:07:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Schreibmayer-W", "name": { "family": "Schreibmayer", "given": "Wolfgang" } }, { "id": "Dessauer-C-W", "name": { "family": "Dessauer", "given": "Carmen W." } }, { "id": "Voroblov-E", "name": { "family": "Voroblov", "given": "Dmitry" } }, { "id": "Gilman-A-G", "name": { "family": "Gilman", "given": "Alfred G." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } } ] }, "title": "Inhibition of an inwardly rectifying K^+ channel by G-protein \u0251-subunits", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1996 Nature Publishing Group. Received 18 October 1995; accepted 5 February 1996.\n\nWe thank E. Peralta and J. P. Adelman for the cDNAs of m2 receptor and rcKATP (Kir3.4), respectively, B. Posner for providing G1i;-c685, and M. Malca for oocyte injections.\nAntisense ODNs to Xenopus G proteins were prepared by W. S. Marshall and co-workers of Amgen Boulder Inc. This work was supported by the Human Frontiers Scientific Program, Israel-USA Binational Science Foundation, Austrian National Bank, Spezialforschungbereich: Biomembranes and Atherosklerosis (Austria), National Institutes of Health, National Institute of General Medical Sciences, and the National Institute of Mental Health.", "abstract": "CHOLINERGIC muscarinic, serotonergic, opioid and several other G-protein-coupled neurotransmitter receptors activate inwardly rectifying K^+ channels of the GIRK family, slowing the heartbeat and decreasing the excitability of neuronal cells. Inhibitory modulation of GIRKs by G-protein-coupled receptors may have important implications in cardiac and brain physiology. Previously G_\u03b1 and G_(\u03b2\u03b3) subunits of heterotrimeric G proteins have both been implicated in channel opening, but recent studies attribute this role primarily to the G_(\u03b2\u03b3) dimer that activates GIRKs in a membrane-delimited fashion, probably by direct binding to the channel protein. We report here that free GTP\u03b3S-activated G_(\u03b1il), but not G_(\u03b1i2) or G_(\u03b1i3), potently inhibits G_(\u03b21\u03b32)-induced GIRK activity in excised membrane patches of Xenopus oocytes expressing GIRK1. High-affinity but partial inhibition is produced by G_(\u03b1s)-GTP\u03b3S. G_(\u03b1il)-GTP\u03b3S also inhibits G_(\u03b2l\u03b32)-activated GIRK in atrial myocytes. Antagonistic interactions between G_\u03b1 and G_(\u03b2\u03b3) may be among the mechanisms determining specificity of G protein coupling to GIRKs.", "date": "1996-04-18", "date_type": "published", "publication": "Nature", "volume": "380", "number": "6575", "publisher": "Nature Publishing Group", "pagerange": "624-627", "id_number": "CaltechAUTHORS:20150610-111824247", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150610-111824247", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Human Frontier Science Program" }, { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "Austrian National Bank" }, { "agency": "Spezialforschungbereich Biomembranes and Atherosklerosis (Austria)" }, { "agency": "NIH" }, { "agency": "National Institute of General Medical Sciences" }, { "agency": "National Institute of Mental Health (NIMH)" } ] }, "doi": "10.1038/380624a0", "resource_type": "article", "pub_year": "1996", "author_list": "Schreibmayer, Wolfgang; Dessauer, Carmen W.; et el." }, { "id": "https://authors.library.caltech.edu/records/wfcdz-8zp09", "eprint_id": 29184, "eprint_status": "archive", "datestamp": "2023-08-20 07:19:33", "lastmod": "2023-10-24 18:25:20", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Figl-A", "name": { "family": "Figl", "given": "Antonio" } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } } ] }, "title": "Voltage-Jump Relaxation Kinetics for Wild-type and Chimeric \u03b2 Subunits of Neuronal Nicotinic Receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1996 Rockefeller University Press. Original version received 14 August 1995 and accepted version received 13 November 1995. We thank Yinong Zhang, Michael Quick, Mark Nowak, and Craig Doupnik for helpful suggestions, Purnima\nDeshpande for expert technical help, and Jeremy Gollub, Heather Davis, and Brad Henkle for oocyte preparation and maintenance. This work was supported by grants from the National Institutes of Health (NS-t 1756) and from the California Tobacco-Related Disease Research Program (1 RT-0365, 1RT-0286).\n\nPublished - FIGjgp96.pdf
", "abstract": "We have studied the voltage-jump relaxation currents for a series of neuronal nicotinic acetylcholine receptors resulting from the coexpression of wild-type and chimeric \u03b24/\u03b22 subunits with \u03b13 subunits in Xenopus oocytes. With acetylcholine as the agonist, the wild-type \u03b13\u03b24 receptors displayed five- to eightfold slower voltage-jump relaxations than did the wild-type \u03b13\u03b22 receptors. In both cases, the relaxations could best be described by two exponential components of approximately equal amplitudes over a wide range of [ACh]'s. Relaxation rate constants increased with [ACh] and saturated at 20- to 30-fold lower concentrations for the \u03b13\u03b22 receptor than for the \u03b13\u03b24 receptor, as observed previously for the peak steady state conductance. Furthermore, the chimeric \u03b24/\u03b22 subunits showed a transition in the concentration dependence of the rate constants in the region between residues 94 and 109, analogous to our previous observation with steady state conductances. However, our experiments with a series of \u03b2-subunit chimeras did not localize residues that govern the absolute value of the kinetic parameters. Hill coefficients for the relaxations also differed from those previously measured for steady state responses. The data reinforce previous conclusions that the region between residues 94 and 109 on the \u03b2 subunit plays a role in binding agonist but also show that other regions of the receptor control gating kinetics subsequent to the binding step.", "date": "1996-03", "date_type": "published", "publication": "Journal of General Physiology", "volume": "107", "number": "3", "publisher": "Rockefeller University Press", "pagerange": "369-379", "id_number": "CaltechAUTHORS:20120208-075012754", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120208-075012754", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "1RT-0365" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "1RT-0286" } ] }, "doi": "10.1085/jgp.107.3.369", "pmcid": "PMC2216994", "primary_object": { "basename": "FIGjgp96.pdf", "url": "https://authors.library.caltech.edu/records/wfcdz-8zp09/files/FIGjgp96.pdf" }, "resource_type": "article", "pub_year": "1996", "author_list": "Figl, Antonio; Labarca, Cesar; et el." }, { "id": "https://authors.library.caltech.edu/records/bvaxk-gca10", "eprint_id": 24130, "eprint_status": "archive", "datestamp": "2023-08-20 07:11:57", "lastmod": "2023-10-23 20:19:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Su-Alyce", "name": { "family": "Su", "given": "Alyce" } }, { "id": "Mager-S", "name": { "family": "Mager", "given": "Sela" } }, { "id": "Mayo-S-L", "name": { "family": "Mayo", "given": "Stephen L." }, "orcid": "0000-0002-9785-5018" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "A multi-substrate single-file model for ion-coupled transporters", "ispublished": "pub", "full_text_status": "public", "keywords": "GABA Plasma Membrane Transport Proteins, Carrier Proteins, Biophysics, Serotonin, Monosaccharide Transport Proteins, Molecular Structure, Ion Transport, Electrochemistry, Nerve Tissue Proteins, Models: Biological, Animals, Serotonin Plasma Membrane Transport Proteins, Membrane Glycoproteins, Binding Sites, Membrane Transport Proteins, Organic Anion Transporters, Membrane Potentials, Membrane Proteins, gamma-Aminobutyric Acid, Computer Simulation, Sodium, Sodium-Glucose Transporter 1, Biophysical Phenomena", "note": "\u00a9 1996 The Biophysical Society; Published by Elsevier Inc.\nReceived for publication 25 July 1995 and in final form 21 October 1995.\nAvailable online 2 January 2009. \nWe thank Eric Bax, Bassil Dahiyat, Norman Davidson, and Jun Li for\nsuggestions. Scott Fraser suggested the diffusion pump analogy.\nThis work was supported by grants from the National Institute of Neurological\nDiseases and Stroke and the National Institute on Drug Abuse. SLM\nacknowledges support from the Rita Allen Foundation, the David and\nLucille Packard Foundation, and the Searle Scholars Program.\n\nPublished - Su_1996_Biophys_J_A_multi-substrate_single-file_model_for.pdf
", "abstract": "Ion-coupled transporters are simulated by a model that differs from contemporary alternating-access schemes. Beginning with concepts derived from multi-ion pores, the model assumes that substrates (both inorganic ions and small organic molecules) hop a) between the solutions and binding sites and b) between binding sites within a single-file pore. No two substrates can simultaneously occupy the same site. Rate constants for hopping can be increased both a) when substrates in two sites attract each other into a vacant site between them and b) when substrates in adjacent sites repel each other. Hopping rate constants for charged substrates are also modified by the membrane field. For a three-site model, simulated annealing yields parameters to fit steady-state measurements of flux coupling, transport-associated currents, and charge movements for the GABA transporter GAT1. The model then accounts for some GAT1 kinetic data as well. The model also yields parameters that describe the available data for the rat 5-HT transporter and for the rabbit Na(+)-glucose transporter. The simulations show that coupled fluxes and other aspects of ion transport can be explained by a model that includes local substrate-substrate interactions but no explicit global conformational changes.", "date": "1996-02", "date_type": "published", "publication": "Biophysical Journal", "volume": "70", "number": "2", "publisher": "Biophysical Society", "pagerange": "762-777", "id_number": "CaltechAUTHORS:20110620-160434844", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20110620-160434844", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "National Institute of Neurological Disorders and Stroke (NINDS)" }, { "agency": "National Institute on Drug Abuse" }, { "agency": "Rita Allen Foundation" }, { "agency": "David and Lucille Packard Foundation" }, { "agency": "Searle Scholars Program" } ] }, "doi": "10.1016/S0006-3495(96)79616-9", "pmcid": "PMC1224976", "primary_object": { "basename": "Su_1996_Biophys_J_A_multi-substrate_single-file_model_for.pdf", "url": "https://authors.library.caltech.edu/records/bvaxk-gca10/files/Su_1996_Biophys_J_A_multi-substrate_single-file_model_for.pdf" }, "resource_type": "article", "pub_year": "1996", "author_list": "Su, Alyce; Mager, Sela; et el." }, { "id": "https://authors.library.caltech.edu/records/b2ndf-dwc15", "eprint_id": 58163, "eprint_status": "archive", "datestamp": "2023-08-20 06:12:24", "lastmod": "2023-10-23 19:07:21", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Nowak-M-W", "name": { "family": "Nowak", "given": "Mark W." } }, { "id": "Zhang-Halyun", "name": { "family": "Zhang", "given": "Halyun" } }, { "id": "Tang-Lixin", "name": { "family": "Tang", "given": "Lixin" } }, { "id": "Deshpande-P", "name": { "family": "Deshpande", "given": "Purnima" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Channel gating governed symmetrically by conserved leucine residues in the M2 domain of nicotinic receptors", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1995 Nature Publishing Group. Received 9 March; accepted 31 May 1995.\n\nWe thank A. Auerbach, C. Chavkin, N. Davidson, D. Dougherty, A. Figl, P. Kearney, P. Koluji and Y. Zhang tor advice. This work was supported by grants tram the NIH and from the California Tobacco-Related Disease Research Project and by an NRSA to Mark W. Nowak.", "abstract": "IN nicotinic acetylcholine receptors (nAChR), as well as glycine, GABA_A (\u03b3-aminobutyric acid), serotonin (5-HT_3), and GluCl glut-amate receptors, a leucine residue at the approximate midpoint of the M2 transmembrane domain (the 9\u2032 position) is conserved across most known subunits. Structural data for the nAChR suggest that the Leu 9\u2032 residues occupy a 'kink' in each of the five M2 helices and point into the closed channel; in the opening step, the M2 helices rotate so that Leu 9\u2032 side chains no longer occlude the conduction pathway. Mutation of Leu 9\u2032 to one of several other residues slows desensitization and increases sensitivity to agonist. We have exploited the \u03b1_2\u03b2\u03b3\u03b4 stoichiometry of muscle nAChR to express receptors with m_s^* = 0 to 5 Leu 9\u2032Ser mutated subunits. Strikingly, each Leu 9\u2032Ser mutation shifts the dose-response relation for ACh to the left by \u02dc10-fold; a nAChR with m_s^* = 4 is 10^4-fold more sensitive than the wild type. The results suggest that each of the five Leu 9\u2032 residues participates independently and symmetrically in a key step in the structural transition between the closed and open states.", "date": "1995-08-10", "date_type": "published", "publication": "Nature", "volume": "376", "number": "6540", "publisher": "Nature Publishing Group", "pagerange": "514-516", "id_number": "CaltechAUTHORS:20150610-113831280", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150610-113831280", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "California Tobacco-Related Disease Research Project" }, { "agency": "National Research Service Award" } ] }, "doi": "10.1038/376514a0", "resource_type": "article", "pub_year": "1995", "author_list": "Labarca, Cesar; Nowak, Mark W.; et el." }, { "id": "https://authors.library.caltech.edu/records/64htz-w3g22", "eprint_id": 12188, "eprint_status": "archive", "datestamp": "2023-08-22 10:32:47", "lastmod": "2023-10-17 16:35:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Doupnik-C-A", "name": { "family": "Doupnik", "given": "Craig A." } }, { "id": "Ivanina-T", "name": { "family": "Ivanina", "given": "Tatiana" } }, { "id": "Bausch-S", "name": { "family": "Bausch", "given": "Suzanne" } }, { "id": "Wang-Weizhen", "name": { "family": "Wang", "given": "Weizhen" } }, { "id": "Lin-Catherine-P", "name": { "family": "Lin", "given": "Catherine" } }, { "id": "Garvey-J-S", "name": { "family": "Garvey", "given": "Justine" } }, { "id": "Chavkin-C", "name": { "family": "Chavkin", "given": "Charles" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Inhibition of function in Xenopus oocytes of the inwardly rectifying G-protein-activated atrial K channel (GIRK1) by overexpression of a membrane-attached form of the C-terminal tail", "ispublished": "pub", "full_text_status": "public", "keywords": "POTASSIUM CHANNEL; EXPRESSION; CLONING; PEPTIDE", "note": "\u00a9 1995 by the National Academy of Sciences. \n\nContributed by Norman Davidson, April 7, 1995. \n\nWe thank B. Henkle for oocyte preparations and Dr. T.A. Patterson for assistance in the affinity purification of the anti-GIRK1 antibodies. C.A.D. has been supported by an American Heart Association fellowship. S.B. is supported by a National Research Service Award training grant. Support by research grants from the National Institute of Mental Health, the National Institute of General Medical Sciences, the National Institute on Drug Abuse, the U.S.-Israel Binational Science Foundation, and the International Human Frontier Scientific Programme is also gratefully acknowledged. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - DASpnas95.pdf
", "abstract": "Coexpression in Xenopus oocytes of the inwardly rectifying guanine nucleotide binding (G)-protein-gated K channel GIRK1 with a myristoylated modification of the (putative) cytosolic C-terminal tail [GIRK1 aa 183-501 fused in-frame to aa 1-15 of p60src and denoted src+ (183-501)] leads to a high degree of inhibition of the inward G-protein-gated K+ current. The nonmyristoylated segment, src- (183-501), is not active. Although some interference with assembly is not precluded, the evidence indicates that the main mechanism of inhibition is interference with functional activation of the channel by G proteins. In part, the tail functions as a blocking particle similar to a \"Shaker ball\"; it may also function by competing for the available supply of free G beta gamma liberated by hormone activation of a seven-helix receptor. The non-G-protein-gated weak inward rectifier ROMK1 is less effectively inhibited, and a Shaker K channel was not inhibited. Immunological assays show the presence of a high concentration of src+ (183-501) in the plasma membrane and the absence of any membrane forms for the nonmyristoylated segment.", "date": "1995-07-18", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "92", "number": "15", "publisher": "National Academy of Sciences", "pagerange": "6758-6762", "id_number": "CaltechAUTHORS:DASpnas95", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:DASpnas95", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "American Heart Association" }, { "agency": "National Research Service" }, { "agency": "National Institute of Mental Health (NIMH)" }, { "agency": "National Institute of General Medical Sciences" }, { "agency": "National Institute on Drug Abuse" }, { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "Human Frontier Science Program" } ] }, "doi": "10.1073/pnas.92.15.6758", "pmcid": "PMC41408", "primary_object": { "basename": "DASpnas95.pdf", "url": "https://authors.library.caltech.edu/records/64htz-w3g22/files/DASpnas95.pdf" }, "resource_type": "article", "pub_year": "1995", "author_list": "Dascal, Nathan; Doupnik, Craig A.; et el." }, { "id": "https://authors.library.caltech.edu/records/aq7nh-36581", "eprint_id": 9187, "eprint_status": "archive", "datestamp": "2023-08-22 10:31:40", "lastmod": "2023-10-16 21:57:33", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kofuji-Paulo", "name": { "family": "Kofuji", "given": "Paulo" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Evidence that neuronal G-protein-gated inwardly rectifying K+ channels are activated by G\u03b2\u03b3 subunits and function as heteromultimers", "ispublished": "pub", "full_text_status": "public", "keywords": "DORSAL RAPHE NEURONS; POTASSIUM CHANNELS; RECEPTORS; EXPRESSION; BINDING; HEART; HIPPOCAMPUS; SEROTONIN; CURRENTS; INVITRO", "note": "\u00a9 1995 by the National Academy of Sciences. \n\nContributed by Norman Davidson, April 10, 1995. \n\nWe thank Brad Henkle for preparing the oocytes, Craig Doupnik and Yinong Zhang for advice, and Dr. Michel Lazdunski for sharing a preprint of ref. 22 while it was in press. This work was supported by ICAgen, Inc., and by National Institutes of Health Grants GM29836 and MH49176. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - KOFpnas95.pdf
", "abstract": "Guanine nucleotide-binding proteins (G proteins) activate K+ conductances in cardiac atrial cells to slow heart rate and in neurons to decrease excitability. cDNAs encoding three isoforms of a G-protein-coupled, inwardly rectifying K+ channel (GIRK) have recently been cloned from cardiac (GIRK1/Kir 3.1) and brain cDNA libraries (GIRK2/Kir 3.2 and GIRK3/Kir 3.3). Here we report that GIRK2 but not GIRK3 can be activated by G protein subunits G\u03b21 and G2 in Xenopus oocytes. Furthermore, when either GIRK3 or GIRK2 was coexpressed with GIRK1 and activated either by muscarinic receptors or by G\u03b2 subunits, G-protein-mediated inward currents were increased by 5- to 40-fold. The single-channel conductance for GIRK1 plus GIRK2 coexpression was intermediate between those for GIRK1 alone and for GIRK2 alone, and voltage-jump kinetics for the coexpressed channels displayed new kinetic properties. On the other hand, coexpression of GIRK3 with GIRK2 suppressed the GIRK2 alone response. These studies suggest that formation of heteromultimers involving the several GIRKs is an important mechanism for generating diversity in expression level and function of neurotransmitter-coupled, inward rectifier K+ channels.", "date": "1995-07-03", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "92", "number": "14", "publisher": "National Academy of Sciences", "pagerange": "6542-6546", "id_number": "CaltechAUTHORS:KOFpnas95", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:KOFpnas95", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "MH49176" }, { "agency": "ICAgen, Inc." } ] }, "doi": "10.1073/pnas.92.14.6542", "pmcid": "PMC41554", "primary_object": { "basename": "KOFpnas95.pdf", "url": "https://authors.library.caltech.edu/records/aq7nh-36581/files/KOFpnas95.pdf" }, "resource_type": "article", "pub_year": "1995", "author_list": "Kofuji, Paulo; Davidson, Norman; et el." }, { "id": "https://authors.library.caltech.edu/records/5dd26-qqj47", "eprint_id": 29337, "eprint_status": "archive", "datestamp": "2023-08-20 06:02:36", "lastmod": "2023-10-24 22:06:55", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Doupnik-C-A", "name": { "family": "Doupnik", "given": "Craig A." } }, { "id": "Lim-Nancy-F", "name": { "family": "Lim", "given": "Nancy F." } }, { "id": "Kofuji-P", "name": { "family": "Kofuji", "given": "Paulo" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Intrinsic Gating Properties of a Cloned G Protein-activated Inward Rectifier K^+ Channel", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1995 Rockefeller University Press. Original version received 2 December 1994 and accepted version received 29 March 1995. Published July 1, 1995. We thank Heather Davis and Brad Henkle for preparation of oocytes. This work was supported by fellowships from the American Heart Association (C. A. Doupnik)\nand the National Institutes of Health (NIH) (N. F. Lim) and by NIH grants GM29836 and MH49176.\n\nPublished - DOUjgp95.pdf
", "abstract": "The voltage-, time-, and K^+-dependent properties of a G protein-activated inwardly rectifying K^+ channel (GIRK1/KGA/Kir3.1) cloned from rat atrium were studied in Xenopus oocytes under two-electrode voltage clamp. During maintained G protein activation and in the presence of high external K^+ (V_K = 0 mV), voltage jumps from V_K to negative membrane potentials activated inward GIRK1 K^+ currents with three distinct time-resolved current components. GIRK1 current activation consisted of an instantaneous component that was followed by two components with time constants T_f~50 ms and T_s~400 ms. These activation time constants were weakly voltage dependent, increasing approximately twofold with maximal hyperpolarization from V_K. Voltage-dependent GIRK1 availability, revealed by tail currents at -80 mV after long prepulses, was greatest at potentials negative to V_K and declined to a plateau of approximately half the maximal level at positive voltages. Voltage-dependent GIRK1 availability shifted with V_K and was half maximal at V_K -20 mV; the equivalent gating charge was ~1.6 e^-. The voltage-dependent gating parameters of GIRK1 did not significantly differ for G protein activation by three heterologously expressed signaling pathways: m2 muscarinic receptors, serotonin 1A receptors, or G protein \u03b21y2 subunits. Voltage dependence was also unaffected by agonist concentration. These results indicate that the voltage-dependent gating properties of GIRK1 are not due to extrinsic factors such as agonist-receptor interactions and G protein-channel coupling, but instead are analogous to the intrinsic gating behaviors of other inwardly rectifying K^+ channels.", "date": "1995-07", "date_type": "published", "publication": "Journal of General Physiology", "volume": "106", "number": "1", "publisher": "Rockefeller University Press", "pagerange": "1-23", "id_number": "CaltechAUTHORS:20120216-103326772", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120216-103326772", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "American Heart Association" }, { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "MH49176" } ] }, "doi": "10.1085/jgp.106.1.1", "pmcid": "PMC2229253", "primary_object": { "basename": "DOUjgp95.pdf", "url": "https://authors.library.caltech.edu/records/5dd26-qqj47/files/DOUjgp95.pdf" }, "resource_type": "article", "pub_year": "1995", "author_list": "Doupnik, Craig A.; Lim, Nancy F.; et el." }, { "id": "https://authors.library.caltech.edu/records/2e1ct-wc395", "eprint_id": 10849, "eprint_status": "archive", "datestamp": "2023-08-22 10:27:26", "lastmod": "2023-10-16 23:08:20", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "B. N." } }, { "id": "Figl-A", "name": { "family": "Figl", "given": "A." } }, { "id": "Quick-M-W", "name": { "family": "Quick", "given": "M. W." } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "C." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Regions of beta 2 and beta 4 responsible for differences between the steady state dose-response relationships of the alpha 3 beta 2 and alpha 3 beta 4 neuronal nicotinic receptors", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1995 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nOriginal version received 25 May 1994 and accepted version received 10 February 1995. \n\nWe thank Jeremy Gollub and Pumima Deshpande for technical assistance in oocyte injection and construction of some chimeras. \n\nThis work was supported by grants from the National Institute of Health (NS-11756 California Tobacco-Related Disease Program (1RT-0365, 1RT-0286).\n\nPublished - COHjgp95.pdf
", "abstract": "We constructed chimeras of the rat beta 2 and beta 4 neuronal nicotinic subunits to locate the regions that contribute to differences between the acetylcholine (ACh) dose-response relationships of the alpha 3 beta 2 and alpha 3 beta 4 receptors. Expressed in Xenopus oocytes, the alpha 3 beta 2 receptor displays an EC50 for ACh approximately 20-fold less than the EC50 of the alpha 3 beta 4 receptor. The apparent Hill slope (n(app)) of alpha 3 beta 2 is near one whereas the alpha 3 beta 4 receptor displays an n(app) near two. Substitutions within the first 120 residues convert the EC50 for ACh from one wild-type value to the other. Exchanging just beta 2:104-120 for the corresponding region of beta 4 shifts the EC50 of ACh dose-response relationship in the expected direction but does not completely convert the EC50 of the dose- response relationship from one wild-type value to the other. However, substitutions in the beta 2:104-120 region do account for the relative sensitivity of the alpha 3 beta 2 receptor to cytisine, tetramethylammonium, and ACh. The expression of beta 4-like (strong) cooperativity requires an extensive region of beta 4 (beta 4:1-301). Relatively short beta 2 substitutions (beta 2:104-120) can reduce cooperativity to beta 2-like values. The results suggest that amino acids within the first 120 residues of beta 2 and the corresponding region of beta 4 contribute to an agonist binding site that bridges the alpha and beta subunits in neuronal nicotinic receptors.", "date": "1995-06", "date_type": "published", "publication": "Journal of General Physiology", "volume": "105", "number": "6", "publisher": "Rockefeller University Press", "pagerange": "745-764", "id_number": "CaltechAUTHORS:COHjgp95", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:COHjgp95", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "1RT-0365" }, { "agency": "California Tobacco-Related Disease Research Program", "grant_number": "1RT-0286" } ] }, "doi": "10.1085/jgp.107.3.369", "pmcid": "PMC2216958", "primary_object": { "basename": "COHjgp95.pdf", "url": "https://authors.library.caltech.edu/records/2e1ct-wc395/files/COHjgp95.pdf" }, "resource_type": "article", "pub_year": "1995", "author_list": "Cohen, B. N.; Figl, A.; et el." }, { "id": "https://authors.library.caltech.edu/records/ap0d7-0hy67", "eprint_id": 53893, "eprint_status": "archive", "datestamp": "2023-08-20 05:45:19", "lastmod": "2023-10-19 22:21:07", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nowak-M-W", "name": { "family": "Nowak", "given": "Mark W." } }, { "id": "Kearney-P-C", "name": { "family": "Kearney", "given": "Patrick C." } }, { "id": "Sampson-J-R", "name": { "family": "Sampson", "given": "Jeffrey R." } }, { "id": "Saks-M-E", "name": { "family": "Saks", "given": "Margaret E." } }, { "id": "Labarca-C-G", "name": { "family": "Labarca", "given": "Cesar G." } }, { "id": "Silverman-S-K", "name": { "family": "Silverman", "given": "Scott K." } }, { "id": "Zhong-Wenge", "name": { "family": "Zhong", "given": "Wenge" } }, { "id": "Thorson-J", "name": { "family": "Thorson", "given": "Jon" } }, { "id": "Abelson-J-N", "name": { "family": "Abelson", "given": "John N." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Schultz-P-G", "name": { "family": "Schultz", "given": "Peter G." } }, { "id": "Dougherty-D-A", "name": { "family": "Dougherty", "given": "Dennis A." }, "orcid": "0000-0003-1464-2461" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Nicotinic Receptor Binding Site Probed with Unnatural Amino Acid Incorporation in Intact Cells", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1995 American Association for the Advancement of Science.\n\nNovember 1994; accepted 23 January 1995.\n\nWe thank V. Cornish, P. Deshpande, O. Uhlenbeck, and Y. Zhang for suggestions; E. Chapman for help with synthesis; and J. Jankowski for help with the measurements. Sponsored by grants from NIH, the Office of Naval Research, the Howard Hughes Medical Institute, the University of California Tobacco-Related Disease Research Project, and the Beckman Institute at Caltech.", "abstract": "The nonsense codon suppression method for unnatural amino acid incorporation has been applied to intact cells and combined with electrophysiological analysis to probe structure-function relations in the nicotinic acetylcholine receptor. Functional receptors were expressed in Xenopus oocytes when tyrosine and phenylalanine derivatives were incorporated at positions 93, 190, and 198 in the binding site of the \u03b1 subunit. Subtle changes in the structure of an individual side chain produced readily detectable changes in the function of this large channel protein. At each position, distinct features of side chain structure dominated the dose-response relation, probably by governing the agonist-receptor binding.", "date": "1995-04-21", "date_type": "published", "publication": "Science", "volume": "268", "number": "5209", "publisher": "American Association for the Advancement of Science", "pagerange": "439-442", "id_number": "CaltechAUTHORS:20150120-140412390", "issn": "0036-8075", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150120-140412390", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "Office of Naval Research (ONR)" }, { "agency": "Howard Hughes Medical Institute (HHMI)" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Caltech Beckman Institute" } ] }, "doi": "10.1126/science.7716551", "resource_type": "article", "pub_year": "1995", "author_list": "Nowak, Mark W.; Kearney, Patrick C.; et el." }, { "id": "https://authors.library.caltech.edu/records/gvqyk-spn15", "eprint_id": 6578, "eprint_status": "archive", "datestamp": "2023-08-22 10:20:14", "lastmod": "2023-10-16 20:24:50", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lim-Nancy-Fidler", "name": { "family": "Lim", "given": "Nancy Fidler" } }, { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "A G protein-gated K channel is activated via beta 2-adrenergic receptors and G beta gamma subunits in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1995 by The Rockefeller University Press \n\nOriginal version received 26 July 1994 and accepted version received 31 October 1994. \n\nWe thank Dr. A. Gilman for G\u03b1sQ227L cDNA, Dr. E. Peralta for the m2R cDNA, and Drs. E. Reuveny, P. Siesinger, A.J. Connolly, and S. R. Coughlin for G protein \u03b21 and \u03b32 subunit cDNAs and J. Gollub and H. Davis for help with oocytes and Y. Zhang for participating in some of the experiments. We thank C. Chavkin, C. Doupnik, L. DiMagno, N. McCarty, S. McDonough, M. Quick, and Y. Uezono for advice. \n\nThis work was supported by grants from the National Institutes of Health (including postdoctoral fellowships to N. Lim), from the US-Israel Binational Science Foundation, and from ICAgen, Inc.\n\nPublished - LIMjgp95.pdf
", "abstract": "In many tissues, inwardly rectifying K channels are coupled to seven- helix receptors via the Gi/Go family of heterotrimeric G proteins. This activation proceeds at least partially via G beta gamma subunits. These experiments test the hypothesis that G beta gamma subunits activate the channel even if released from other classes of heterotrimeric G proteins. The G protein-gated K channel from rat atrium, KGA/GIRK1, was expressed in Xenopus oocytes with various receptors and G proteins. The beta 2-adrenergic receptor (beta 2AR), a Gs-linked receptor, activated large KGA currents when the alpha subunit, G alpha s, was also overexpressed. Although G alpha s augmented the coupling between beta 2AR and KGA, G alpha s also inhibited the basal, agonist-independent activity of KGA. KGA currents stimulated via beta 2AR activated, deactivated, and desensitized more slowly than currents stimulated via Gi/Go-linked receptors. There was partial occlusion between currents stimulated via beta 2AR and the m2 muscarinic receptor (a Gi/Go-linked receptor), indicating some convergence in the mechanism of activation by these two receptors. Although stimulation of beta 2AR also activates adenylyl cyclase and protein kinase A, activation of KGA via beta 2AR is not mediated by this second messenger pathway, because direct elevation of intracellular cAMP levels had no effect on KGA currents. Experiments with other coexpressed G protein alpha and beta gamma subunits showed that (a) a constitutively active G alpha s mutant did not suppress basal KGA currents and was only partially as effective as wild type G alpha s in coupling beta 2AR to KGA, and (b) beta gamma subunits increased basal KGA currents. These results reinforce present concepts that beta gamma subunits activate KGA, and also suggest that beta gamma subunits may provide a link between KGA and receptors not previously known to couple to inward rectifiers.", "date": "1995-03", "date_type": "published", "publication": "Journal of General Physiology", "volume": "105", "number": "3", "publisher": "Rockefeller University Press", "pagerange": "421-439", "id_number": "CaltechAUTHORS:LIMjgp95", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LIMjgp95", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH Postdoctoral Fellowship" }, { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "ICAgen, Inc." } ] }, "doi": "10.1085/jgp.105.3.421", "pmcid": "PMC2216943", "primary_object": { "basename": "LIMjgp95.pdf", "url": "https://authors.library.caltech.edu/records/gvqyk-spn15/files/LIMjgp95.pdf" }, "resource_type": "article", "pub_year": "1995", "author_list": "Lim, Nancy Fidler; Dascal, Nathan; et el." }, { "id": "https://authors.library.caltech.edu/records/cjaee-c9b24", "eprint_id": 10182, "eprint_status": "archive", "datestamp": "2023-08-22 10:09:23", "lastmod": "2023-10-16 22:43:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Quick-M-W", "name": { "family": "Quick", "given": "Michael W." } }, { "id": "Simon-M-I", "name": { "family": "Simon", "given": "Melvin I." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Aragay-A-M", "name": { "family": "Aragay", "given": "Anna M." } } ] }, "title": "Differential coupling of G protein alpha subunits to seven-helix receptors expressed in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1994 by the American Society for Biochemistry and Molecular Biology. \n\n(Received for publication, May 13, 1994, and in revised form, September 12, 1994) \n\nWe thank Heather Davis and Jeremy Gollub for oocyte preparation and Dr. Janis Corey for comments on previous versions of the manuscript. CFTR DNA was a gift of Dr. John Riordan (Hospital for Sick Children, Toronto). \n\nThis work was supported in part by National Institutes of Health Grants GM-29836 and MH-49716 and by a postdoctoral fellowship (to M.W.Q). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. \n\nThe nucleotide sequence(s) reported in this paper has been submitted to the GenBank(TM)/EMBL Data Bank with accession number(s) L05540.\n\nPublished - QUIjbc94.pdf
", "abstract": "Xenopus oocytes were used to examine the coupling of the serotonin 1c (5HT1c) and thyrotropin-releasing hormone (TRH) receptors to both endogenous and heterologously expressed G protein alpha subunits. Expression of either G protein-coupled receptor resulted in agonist- induced, Ca(2+)-activated Cl- currents that were measured using a two- electrode voltage clamp. 5HT-induced Cl- currents were reduced 80% by incubating the injected oocytes with pertussis toxin (PTX) and inhibited 50-65% by injection of antisense oligonucleotides to the PTX- sensitive Go alpha subunit. TRH-induced Cl- currents were reduced only 20% by PTX treatment but were inhibited 60% by injection of antisense oligonucleotides to the PTX-insensitive Gq alpha subunit. Injection of antisense oligonucleotides to a novel Xenopus phospholipase C-beta inhibited the 5HT1c (and Go)-induced Cl- current with little effect on the TRH (and Gq)-induced current. These results suggest that receptor- activated Go and Gq interact with different effectors, most likely different isoforms of phospholipase C-beta. Co-expression of each receptor with seven different mammalian G protein alpha subunit cRNAs (Goa, Gob, Gq, G11, Gs, Golf, and Gt) was also examined. Co-expression of either receptor with the first four of these G alpha subunits resulted in a maximum 4-6-fold increase in Cl- currents; the increase depended on the amount of G alpha subunit cRNA injected. This increase was blocked by PTX for G alpha oa and G alpha ob co-expression but not for G alpha q or G alpha 11 co-expression. Co-expression of either receptor with Gs, Golf, or Gt had no effect on Ca(2+)-activated Cl- currents; furthermore, co-expression with Gs or Golf also failed to reveal 5HT- or TRH-induced changes in adenylyl cyclase as assessed by activation of the cystic fibrosis transmembrane conductance regulator Cl- channel. These results indicate that in oocytes, the 5HT1c and TRH receptors do the following: 1) preferentially couple to PTX-sensitive (Go) and PTX-insensitive (Gq) G proteins and that these G proteins act on different effectors, 2) couple within the same cell type to several different heterologously expressed G protein alpha subunits to activate the oocyte's endogenous Cl- current, and 3) fail to couple to G protein alpha subunits that activate cAMP or phosphodiesterase.", "date": "1994-12-02", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "269", "number": "48", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "30164-30172", "id_number": "CaltechAUTHORS:QUIjbc94", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:QUIjbc94", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "MH-49716" }, { "agency": "NIH Postdoctoral Fellowship" } ] }, "primary_object": { "basename": "QUIjbc94.pdf", "url": "https://authors.library.caltech.edu/records/cjaee-c9b24/files/QUIjbc94.pdf" }, "resource_type": "article", "pub_year": "1994", "author_list": "Quick, Michael W.; Simon, Melvin I.; et el." }, { "id": "https://authors.library.caltech.edu/records/wea8w-cgv25", "eprint_id": 893, "eprint_status": "archive", "datestamp": "2023-08-22 10:03:02", "lastmod": "2023-10-23 16:54:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Bradley-J", "name": { "family": "Bradley", "given": "Jonathan" } }, { "id": "Li-Jun", "name": { "family": "Li", "given": "Jun" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Zinn-K", "name": { "family": "Zinn", "given": "Kai" }, "orcid": "0000-0002-6706-5605" } ] }, "title": "Heteromeric olfactory cyclic nucleotide-gated channels: a subunit that confers increased sensitivity to cAMP", "ispublished": "pub", "full_text_status": "public", "keywords": "OLFACTION, CGMP, RECEPTOR-CELLS, ACTIVATED CHANNEL, ODORANT DETECTION, EPITHELIUM, TRANSDUCTION, EXPRESSION, NEURONS, IDENTIFICATION, PHOTORECEPTORS, AMPLIFICATION", "note": "Copyright \u00a9 1994 by the National Academy of Sciences. \n\nContributed by Norman Davidson, May 20, 1994. \n\nWe thank T.-Y. Chen and K.-W. Yau for helpful discussions, HEK 293 cells, and an rOCNC1 clone; W. Zagotta for providing the sequences of the CN1 and CN2 primers; Genentech for the pCIS plasmid; M. Quick and Y. Uezono of Caltech for experimental contributions to the early phase of this project; and L. Buck and E. Liman for communicating data before publication. J.B. was supported by a National Institutes of Health graduate training grant and an Achievement Award for College Scientists. This work was supported by grants from the National Institute of Mental Health and the National Institutes of Health to K.Z., H.A.L., and N.D.\n\nPublished - BRApnas94.pdf
", "abstract": "Olfactory receptor neurons respond to odorant stimulation with a rapid increase in intracellular cAMP that opens cyclic nucleotide-gated (cng) cation channels. cng channels in rat olfactory neurons are activated by cAMP in the low micromolar range and are outwardly rectifying. The cloned rat olfactory cng channel (rOCNC1), however, is much less sensitive to cAMP and exhibits very weak rectification. Here we describe the cloning and characterization of a second rat cng channel subunit, denoted rOCNC2. rOCNC2 does not form functional channels when expressed alone. When rOCNC1 and rOCNC2 are coexpressed, however, an outwardly rectifying cation conductance with cAMP sensitivity near that of the native channel is observed. In situ hybridization with probes specific for the two subunits shows that they are coexpressed in olfactory receptor neurons. These data indicate that the native olfactory cng channel is likely to be a heterooligomer of the rOCNC1 and rOCNC2 subunits.", "date": "1994-09-13", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "91", "number": "19", "publisher": "National Academy of Sciences", "pagerange": "8890-8894", "id_number": "CaltechAUTHORS:BRApnas94", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:BRApnas94", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "National Institute of Mental Health (NIMH)" }, { "agency": "Achievement Award for College Scientists" } ] }, "pmcid": "PMC44712", "primary_object": { "basename": "BRApnas94.pdf", "url": "https://authors.library.caltech.edu/records/wea8w-cgv25/files/BRApnas94.pdf" }, "resource_type": "article", "pub_year": "1994", "author_list": "Bradley, Jonathan; Li, Jun; et el." }, { "id": "https://authors.library.caltech.edu/records/8vrh5-tyb81", "eprint_id": 76393, "eprint_status": "archive", "datestamp": "2023-10-09 23:49:09", "lastmod": "2023-10-24 18:31:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Karschin-C", "name": { "family": "Karschin", "given": "Christine" } }, { "id": "Schreibmayer-W", "name": { "family": "Schreibmayer", "given": "Wolfgang" } }, { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry" }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Karschin-A", "name": { "family": "Karschin", "given": "Andreas" } } ] }, "title": "Distribution and localization of a G protein-coupled inwardly rectifying K\u207a channel in the rat", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Potassium channel; Inward rectifier; G-protein gating; mRNA; Hybridization (in situ)", "note": "\u00a9 1994 Federation of European Biochemical Societies. \n\nManuscript received: 07 April 1994. Manuscript revised: 31 May 1994. \n\nWe wish to thank Dr. Walter St\u00fchmer for hisgenerous support. The study was supported in part by grants from the US National Institutes of Health (GM-29836, MH-49176), the US-Israel Binational Science Foundation, the Austrian Research Foundation and the Human Frontiers Organization.", "abstract": "The cellular distribution of the mRNA of the inwardly rectifying K^+ channel KGA (GIRK1) was investigated in rat tissue by in situ hybridization. KGA was originally cloned from the heart and represents the first G protein\u2010activated K^+ channel identified. It is expressed in peripheral tissue solely in the atrium, but not in the ventricle, skeletal muscle, lung and kidney. In the central nervous system KGA is most prominently expressed in the Ammon's horn and dentate gyrus of the hippocampus, neocortical layers II\u2013VI, cerebellar granular layer, olfactory bulb, anterior pituitary, thalamic nuclei and several distinct nuclei of the lower brainstem. The abundant expression of KGA in many CNS neurons support its important role as a major target channel for G protein mediated receptor function.", "date": "1994-07-11", "date_type": "published", "publication": "FEBS Letters", "volume": "348", "number": "2", "publisher": "Wiley", "pagerange": "139-144", "id_number": "CaltechAUTHORS:20170408-205029469", "issn": "0014-5793", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170408-205029469", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "MH-49176" }, { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "FWF Der Wissenschaftsfonds" }, { "agency": "Human Frontier Science Program" } ] }, "doi": "10.1016/0014-5793(94)00590-7", "resource_type": "article", "pub_year": "1994", "author_list": "Karschin, Christine; Schreibmayer, Wolfgang; et el." }, { "id": "https://authors.library.caltech.edu/records/x3nkn-7e556", "eprint_id": 11882, "eprint_status": "archive", "datestamp": "2023-08-22 09:54:05", "lastmod": "2023-10-17 15:57:38", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Corey-J-L", "name": { "family": "Corey", "given": "Janis L." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Brecha-N", "name": { "family": "Brecha", "given": "Nicholas" } }, { "id": "Quick-M-W", "name": { "family": "Quick", "given": "Michael W." } } ] }, "title": "Protein kinase C modulates the activity of a cloned gamma-aminobutyric acid transporter expressed in Xenopus oocytes via regulated subcellular redistribution of the transporter", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1994 by the American Society for Biochemistry and Molecular Biology. \n\n(Received for publication, December 29, 1993, and in revised form, March 18, 1994) \n\nWe thank M. Stallcup and J. Guastella for valuable discussions and suggestions, and J. Gollub and Heather Davis for oocyte preparation. \n\nThis research was supported by United States Public Health Service Grant NS-11756 and National Research Service Award fellowships (to J.L.C. and M.W.Q.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.\n\nPublished - CORjbc94.pdf
", "abstract": "We report that activators and inhibitors of protein kinase C (PKC) and protein phosphatases regulate the activity of a cloned rat brain gamma- aminobutyric acid (GABA) transporter (GAT1) expressed in Xenopus oocytes. Four compounds known to activate PKC increased GABA uptake 2- 3.5-fold over basal control levels. Inhibition of PKC by bisindolylmaleimide reduced basal GABA uptake 80% and blocked the phorbol 12-myristate 13-acetate (PMA)-induced stimulation of transport. Okadaic acid, a protein phosphatase inhibitor, stimulated transport 2.5- fold; a 4-fold increase in GABA uptake occurred when oocytes were treated with cyclosporin A, a specific inhibitor of protein phosphatase 2B. Modulation resulted in changes to Vmax but not to Km and was influenced by the functional expression level of the transporter protein; as expression level increased, the ability to up-regulate transporter activity decreased. Down-regulation of transporter activity was independent of expression level. Modulation did not occur through phosphorylation of the three consensus PKC sites predicted by the primary protein sequence since their removal had no effect on the susceptibility of the transporter to modulation by PMA or bisindolylmaleimide. Subcellular fractionation of oocyte membranes demonstrated that under basal level conditions, the majority of GAT1 was targeted to a cytoplasmic compartment corresponding to the trans- Golgi or low density vesicles. Stimulation of PKC with PMA resulted in a translocation of transporters from this compartment to the plasma membrane. At higher expression levels of GAT1 protein, a larger portion of GAT1 was found on the plasma membrane during basal level conditions and treatment with bisindolylmaleimide resulted in removal of these transporters from the plasma membrane. At expression levels demonstrated to be resistant to modulation by PMA, PMA-treatment still resulted in translocation of transporters from the cytoplasm to the plasma membrane. Thus, the inability of PMA to increase uptake at high expression of the GAT1 protein is due to saturation at a step subsequent to translocation. These findings 1) demonstrate the presence of a novel regulated secretory pathway in oocytes and 2) suggest a modulatory mechanism for neurotransmitter transporters that could have significant effects upon synaptic function.", "date": "1994-05-20", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "269", "number": "20", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "14759-14767", "id_number": "CaltechAUTHORS:CORjbc94", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:CORjbc94", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH Postdoctoral Fellowship" } ] }, "primary_object": { "basename": "CORjbc94.pdf", "url": "https://authors.library.caltech.edu/records/x3nkn-7e556/files/CORjbc94.pdf" }, "resource_type": "article", "pub_year": "1994", "author_list": "Corey, Janis L.; Davidson, Norman; et el." }, { "id": "https://authors.library.caltech.edu/records/k4w86-8m759", "eprint_id": 29415, "eprint_status": "archive", "datestamp": "2023-08-20 04:06:52", "lastmod": "2023-10-24 22:10:30", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Mager-S", "name": { "family": "Mager", "given": "Sela" } }, { "id": "Quick-M-W", "name": { "family": "Quick", "given": "Michael W." } }, { "id": "Corey-J-L", "name": { "family": "Corey", "given": "Janis L." } } ] }, "title": "Permeation Properties of Neurotransmitter Transporters", "ispublished": "pub", "full_text_status": "restricted", "keywords": "synaptic transmission, electrophysiology, GABA, serotonin", "note": "\u00a9 1994 Annual Reviews.\n\nWe thank D Atwell, B Kanner, G Rudnick, and E Schwartz for helpful\ncomments on the manuscript and N Davidson for stimulating discussions.\nOur research is supported by fellowships from the NIH (J Corey, M Quick),\nthe Muscular Dystrophy Association (S Mager), and the Lester Deutsch\nFoundation (S Mager) and by a grant from the NIH (NS-11756).", "abstract": "The neurotransmitter transporters belong to three known families of intrinsic\nmembrane proteins (Figure 1). The energy for transport, which is often\nagainst the neurotransmitter concentration gradient, is derived from the\ncotransport (and in some cases the counter transport) of inorganic ions.\nIon-transporting ATPases establish the concentration gradients for these ions.", "date": "1994-04", "date_type": "published", "publication": "Annual Review of Pharmacology and Toxicology", "volume": "34", "publisher": "Annual Reviews", "pagerange": "219-249", "id_number": "CaltechAUTHORS:20120222-112704352", "issn": "0362-1642", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120222-112704352", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Lester Deutsch Foundation" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1146/annurev.pharmtox.34.1.219", "resource_type": "article", "pub_year": "1994", "author_list": "Lester, Henry A.; Mager, Sela; et el." }, { "id": "https://authors.library.caltech.edu/records/d7t6y-s1x82", "eprint_id": 52541, "eprint_status": "archive", "datestamp": "2023-08-20 03:56:01", "lastmod": "2023-10-18 20:57:37", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Corey-J-L", "name": { "family": "Corey", "given": "Janis L." } }, { "id": "Quick-M-W", "name": { "family": "Quick", "given": "Michael W." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Guastella-J", "name": { "family": "Guastella", "given": "John" } } ] }, "title": "A cocaine-sensitive Drosophila serotonin transporter: Cloning, expression, and electrophysiological characterization", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1993 National Academy of Sciences.\n\nContributed by Norman Davidson, October 1, 1993.\n\nWe thank Bruce Hamilton for providing us with the Drosophila\nhead library, Jonathan Bradley for Drosophila embryo RNA, Susan R. Halsell for providing Drosophila for RNA preparation, and E. Davidson for use of a DNA sequenator. This research was supported by grants from the U.S. Public Health Service (Grant NS-11756 and National Research Service Award Fellowships to J.L.C., M.W.Q., and J.G.).\n\nPublished - PNAS-1994-Corey-1188-92.pdf
", "abstract": "A cocaine-sensitive, high-affinity Drosophila serotonin (5-hydroxytryptamine; 5HT) transporter cDNA, denoted dSERT1, was isolated and characterized in oocytes. dSERT1 shows little transport of other monoamines and is Na^+ and Cl^- dependent. Sequence analysis indicates 12 putative transmembrane domains and strong homologies (\u224850%) among dSERT1 and mammalian 5HT, norepinephrine, and dopamine transporters. Interestingly, the pharmacological properties of dSERT1, including sensitivity to antidepressants, are more similar to those of mammalian catecholamine transporters than to mammalian 5HT transporters. Two-electrode voltage-clamp analysis demonstrated 5HT-induced, voltage-dependent currents. Cloning and characterization of dSERT1 adds significantly to our knowledge of the diversity of 5HT transporters with regard to primary sequence, pharmacological profile, and permeation properties.", "date": "1994-02-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "91", "number": "3", "publisher": "National Academy of Sciences", "pagerange": "1188-1192", "id_number": "CaltechAUTHORS:20141210-095231086", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20141210-095231086", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH Postdoctoral Fellowship" } ] }, "pmcid": "PMC521479", "primary_object": { "basename": "PNAS-1994-Corey-1188-92.pdf", "url": "https://authors.library.caltech.edu/records/d7t6y-s1x82/files/PNAS-1994-Corey-1188-92.pdf" }, "resource_type": "article", "pub_year": "1994", "author_list": "Corey, Janis L.; Quick, Michael W.; et el." }, { "id": "https://authors.library.caltech.edu/records/ef303-m9a68", "eprint_id": 12194, "eprint_status": "archive", "datestamp": "2023-08-22 09:37:09", "lastmod": "2023-10-17 16:35:33", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Schreibmayer-W", "name": { "family": "Schreibmayer", "given": "Wolfgang" } }, { "id": "Lim-Nancy-F", "name": { "family": "Lim", "given": "Nancy F." } }, { "id": "Wang-Weizhen", "name": { "family": "Wang", "given": "Weizhen" } }, { "id": "Chavkin-C", "name": { "family": "Chavkin", "given": "Charles" } }, { "id": "DiMagno-L", "name": { "family": "DiMagno", "given": "Lisa" } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Kieffer-B-L", "name": { "family": "Kieffer", "given": "Brigitte L." } }, { "id": "Gaveriaux-Ruff-C", "name": { "family": "Gaveriaux-Ruff", "given": "Claire" } }, { "id": "Trollinger-D", "name": { "family": "Trollinger", "given": "David" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Atrial G protein-activated K+ channel: expression cloning and molecular properties", "ispublished": "pub", "full_text_status": "public", "keywords": "GTP-BINDING PROTEINS; DELTA-OPIOID RECEPTOR; ION CHANNELS; MUSCARINIC RECEPTORS; GAMMA-SUBUNIT; CELLS; SEROTONIN; HEART; HIPPOCAMPUS; MEMBRANE", "note": "\u00a9 1993 by the National Academy of Sciences. \n\nContributed by Norman Davidson, July 28, 1993. \n\nWe thank Jun Li for preparing some of the cDNAs and P. Hartig, M. I. Simon, and E. Peralta for supplying the cDNAs of 5HT1A receptor, Gi2 \u03b1 subunit, and m2 receptor, respectively. This work was supported by Public Health Service Grants GM29836, MH49176, and DA04123 (C.C.), by the U.S.-Israel Binational Science Foundation, and by the Austrian Research Foundation. \n\nThe sequences reported in this paper have been deposited in the GenBank database (accession nos. U01071 for KGA and U01141 for KGB1). \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - DASpnas93b.pdf
", "abstract": "Activity of several ion channels is controlled by heterotrimeric GTP-binding proteins (G proteins) via a membrane-delimited pathway that does not involve cytoplasmic intermediates. The best studied example is the K+ channel activated by muscarinic agonists in the atrium, which plays a crucial role in regulating the heartbeat. To enable studies of the molecular mechanisms of activation, this channel, denoted KGA, was cloned from a rat atrium cDNA library by functional coupling to coexpressed serotonin type 1A receptors in Xenopus oocytes. KGA displays regions of sequence homology to other inwardly rectifying channels as well as unique regions that may govern G-protein interaction. The expressed KGA channel is activated by serotonin 1A, muscarinic m2, and delta-opioid receptors via G proteins. KGA is activated by guanosine 5'-[gamma-thio]triphosphate in excised patches, confirming activation by a membrane-delimited pathway, and displays a conductance equal to that of the endogenous channel in atrial cells. The hypothesis that similar channels play a role in neuronal inhibition is supported by the cloning of a nearly identical channel (KGB1) from a rat brain cDNA library.", "date": "1993-11-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "90", "number": "21", "publisher": "National Academy of Sciences", "pagerange": "10235-10239", "id_number": "CaltechAUTHORS:DASpnas93b", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:DASpnas93b", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "MH49176" }, { "agency": "NIH", "grant_number": "DA04123" }, { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "FWF Der Wissenschaftsfonds" } ] }, "doi": "10.1073/pnas.90.21.10235", "pmcid": "PMC47749", "primary_object": { "basename": "DASpnas93b.pdf", "url": "https://authors.library.caltech.edu/records/ef303-m9a68/files/DASpnas93b.pdf" }, "resource_type": "article", "pub_year": "1993", "author_list": "Dascal, Nathan; Schreibmayer, Wolfgang; et el." }, { "id": "https://authors.library.caltech.edu/records/mjzmn-yx925", "eprint_id": 29633, "eprint_status": "archive", "datestamp": "2023-08-20 03:13:36", "lastmod": "2023-10-24 22:20:49", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Slepak-V-Z", "name": { "family": "Slepak", "given": "Vladlen Z." } }, { "id": "Quick-M-W", "name": { "family": "Quick", "given": "Michael W." } }, { "id": "Aragay-A-M", "name": { "family": "Aragay", "given": "Anna M." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Simon-M-I", "name": { "family": "Simon", "given": "Melvin I." } } ] }, "title": "Random Mutagenesis of G protein \u0251 Subunit G_o\u0251. Mutations altering nucleotide binding", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1993 by The American Society for Biochemistry and Molecular Biology, Inc. Received for publication, May 29, 1993, and in revised form, July 7, 1993. \nThe costs of publication of this article were defrayed in part by the\npayment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to\nindicate this fact.\n\nPublished - SLEjbc93b.pdf
", "abstract": "Nucleotide binding properties of the G protein \u0251 subunit G_o\u0251 were probed by mutational analysis in recombinant Escherichia coli. Thousands of random mutations generated by polymerase chain reaction were screened by in situ [^(35)S]GTPyS (guanosine 5'-(3-O-thio)-triphosphate) binding on the colony lifts following transformation of bacteria with modified G_o\u0251 cDNA. Clones that did not bind the nucleotide under these conditions were characterized by DNA sequence analysis, and the nucleotide binding properties were further studied in crude bacterial extracts. A number of novel mutations reducing the affinity of G_o\u0251 for GTPyS or Mg^(2+) were identified. Some of the mutations substitute amino acid residues homologous to those known to interact with guanine nucleotides in p21^(ras) proteins. Other mutations show that previously unstudied residues also participate in the nucleotide binding. Several mutants lost GTPyS binding but retained the capacity to interact with the \u03b2y subunit complex as determined by pertussis toxin-mediated ADP-ribosylation. One of these, mutant S47C, was functionally expressed in Xenopus laevis oocytes along with the G protein-coupled thyrotropin-releasing hormone (TRH) receptor. Whereas wild-type G_o\u0251 increased TRH-promoted chloride currents, S47C significantly decreased the hormone-induced Cl^- response, suggesting that this mutation resulted in a dominant negative phenotype.", "date": "1993-10-15", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "268", "number": "29", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "21889-21894", "id_number": "CaltechAUTHORS:20120307-154341054", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120307-154341054", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "primary_object": { "basename": "SLEjbc93b.pdf", "url": "https://authors.library.caltech.edu/records/mjzmn-yx925/files/SLEjbc93b.pdf" }, "resource_type": "article", "pub_year": "1993", "author_list": "Slepak, Vladlen Z.; Quick, Michael W.; et el." }, { "id": "https://authors.library.caltech.edu/records/e7jk6-x6t37", "eprint_id": 58129, "eprint_status": "archive", "datestamp": "2023-08-20 03:03:00", "lastmod": "2023-10-23 19:05:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } } ] }, "title": "The response to vagusstoff", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1993 Nature Publishing Group.", "abstract": "The molecular cloning of all the major components in the transduction pathway described in Otto Loewi's famous experiments on isolated frog hearts' is now complete with the publication of a letter by Kubo et al. on page 802 of this issue. Loewi cannulated two hearts and connected\nthe effluent from heart A to the input of heart B. He then stimulated the vagus nerve to heart A; in agreement with\nprevious experiments, heart A ceased beating for several minutes.", "date": "1993-08-26", "date_type": "published", "publication": "Nature", "volume": "364", "number": "6440", "publisher": "Nature Publishing Group", "pagerange": "758-759", "id_number": "CaltechAUTHORS:20150609-125940793", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150609-125940793", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1038/364758a0", "resource_type": "article", "pub_year": "1993", "author_list": "Lester, Henry A. and Dascal, Nathan" }, { "id": "https://authors.library.caltech.edu/records/7nsa0-14487", "eprint_id": 10662, "eprint_status": "archive", "datestamp": "2023-08-22 09:28:33", "lastmod": "2023-10-16 23:01:36", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Lim-Nancy-F", "name": { "family": "Lim", "given": "Nancy F." } }, { "id": "Schreibmayer-W", "name": { "family": "Schreibmayer", "given": "Wolfgang" } }, { "id": "Wang-Weizhen", "name": { "family": "Wang", "given": "Weizhen" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Expression of an atrial G-protein-activated potassium channel in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "keywords": "MUSCARINIC K+ CHANNELS; GTP-BINDING PROTEINS; BETA-GAMMA-SUBUNITS; MESSENGER-RNA; ACETYLCHOLINE ACTIVATION; ALPHA-SUBUNITS; ION CHANNELS; RECEPTORS; CELLS; SEROTONIN", "note": "\u00a9 1993 by the National Academy of Sciences. \n\nContributed by Norman Davidson, April 9, 1993. \n\nWe thank P. Hartig and M.I. Simon for providing the cDNAs of SHT1A-R and Gi2\u03b1. This work was supported by U.S. Public Health Service Grants GM-29836 and MH-49176, by U.S.-Israel Binational Science Foundation, a National Research Service Award, and Austrian Research Foundation. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - DASpnas93.pdf
", "abstract": "Injection of rat atrial RNA into Xenopus oocytes resulted in the expression of a guanine nucleotide binding (G) protein-activated K+ channel. Current through the channel could be activated by acetylcholine or, if RNA encoding a neuronal 5HT1A receptor was coinjected with atrial RNA, by serotonin (5HT). A 5HT-evoked current (I5HT) was observed in oocytes injected with ventricle RNA fractions (of 2.5-5.5 kb) and 5HT1A receptor RNA. I5HT displayed strong inward rectification with very little conductance above the K+ equilibrium potential, was highly selective for K+ over Na+, and was blocked by 5-300 \u00b5M Ba2+. I5HT was suppressed by intracellular injection of the nonhydrolyzable analog of GDP, guanosine 5'-[\u00df-thio]diphosphate, but not by treatment with pertussis toxin (PTX), suggesting coupling of the receptor to the G-protein-activated K+ channel via a PTX-insensitive G protein, possibly endogenously present in the oocyte. Coexpression of the subunit of a PTX-sensitive G protein, Gi2, rendered I5HT sensitive to PTX inhibition. Native oocytes displayed a constitutively active inwardly rectifying K+ current with a lower sensitivity to Ba2+ block; expression of a similar current was also directed by atrial or ventricle RNA of 1.5-3 kb. Xenopus oocytes may be employed for cloning of the G-protein-activated K+ channel cDNA and for studying the coupling between this channel and G proteins.", "date": "1993-07-15", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "90", "number": "14", "publisher": "National Academy of Sciences", "pagerange": "6596-6600", "id_number": "CaltechAUTHORS:DASpnas93", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:DASpnas93", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1073/pnas.90.14.6596", "pmcid": "PMC46979", "primary_object": { "basename": "DASpnas93.pdf", "url": "https://authors.library.caltech.edu/records/7nsa0-14487/files/DASpnas93.pdf" }, "resource_type": "article", "pub_year": "1993", "author_list": "Dascal, Nathan; Lim, Nancy F.; et el." }, { "id": "https://authors.library.caltech.edu/records/zn4rp-png14", "eprint_id": 29664, "eprint_status": "archive", "datestamp": "2023-08-20 02:52:31", "lastmod": "2023-10-24 22:21:45", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "McCarthy-N-A", "name": { "family": "McCarthy", "given": "N. A." } }, { "id": "McDonough-S", "name": { "family": "McDonough", "given": "S." } }, { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "B. N." } }, { "id": "Riordan-J-R", "name": { "family": "Riordan", "given": "J. R." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Voltage-dependent Block of the Cystic Fibrosis Transmembrane Conductance Regulator Cl- Channel by Two Closely\n Related Arylaminobenzoates", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1993 Rockefeller University Press. Original version received 7January 1993 and accepted version received 17 March 1993. Published July 1, 1993.\nThe authors wish to thank Dr. Xian-cheng Yang for advice on single-channel analysis, Prof. Anita Zimmerman and Prof. Dennis Dougherty for helpful discussions, Dr. Michael Quick for assistance, and Dr. B. K. Kobilka and colleagues (Dept. of Cellular and Molecular Physiology, Stanford University Medical Center, Stanford, CA) for supplying us with the \u03b22-AR cDNA. \nThis project was supported by grants from the Cystic Fibrosis Foundation (Z139) and from the NIH (GM-29836). N. A. McCarty was supported by an NIH NRSA fellowship (DK08559) and S.\nMcDonough was supported by an NIH training grant (GM-07737).\n\nPublished - MCCjgp93.pdf
", "abstract": "The gene defective in cystic fibrosis encodes a Cl- channel, the cystic fibrosis transmembrane conductance regulator (CFTR). CFTR is blocked by diphenylamine-2-carboxylate (DPC) when applied extracellularly at millimolar concentrations. We studied the block of CFTR expressed in Xenopus oocytes by DPC or by a closely related molecule, flufenamic acid (FFA). Block of whole-cell CFTR currents by bath-applied DPC or by FFA, both at 200 \u00b5M, requires several minutes to reach full effect. Blockade is voltage dependent, suggesting open-channel block: currents at positive potentials are not affected but currents at negative potentials are reduced. The binding site for both drugs senses ~40% of the electric field across the membrane, measured from the inside. In single-channel recordings from excised patches without blockers, the conductance was 8.0 \u00b1 0.4 pS in symmetric 150 mM Cl^-. A subconductance state, measuring ~60% of the main conductance, was often observed. Bursts to the full open state lasting up to tens of seconds were uninterrupted at depolarizing membrane voltages. At hyperpolarizing voltages, bursts were interrupted by brief closures. Either DPC or FFA (50 \u00b5M) applied to the cytoplasmic or extracellular face of the channel led to an increase in flicker at V_m =-100 mV and not at V_m = +100 mV, in agreement with whole-cell experiments. DPC induced a higher frequency of flickers from the cytoplasmic side than the extracellular side. FFA produced longer closures than DPC; the FFA closed time was roughly equal (~ 1.2 ms) at -100 mV with application from either side. In cell-attached patch recordings with DPC or FFA applied to the bath, there was flickery block at V_m = -100 mV, confirming that the drugs permeate through the membrane to reach the binding site. The data are consistent with the presence of a single binding site for both drugs, reached from either end of the channel. Open-channel block by DPC or FFA may offer tools for use with site-directed mutagenesis to describe the permeation pathway.", "date": "1993-07", "date_type": "published", "publication": "Journal of General Physiology", "volume": "102", "number": "1", "publisher": "Rockefeller University Press", "pagerange": "1-23", "id_number": "CaltechAUTHORS:20120309-113518954", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120309-113518954", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Cystic Fibrosis Foundation", "grant_number": "Z139" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "DK08559" }, { "agency": "NIH", "grant_number": "GM-07737" } ] }, "doi": "10.1085/jgp.102.1.1", "pmcid": "PMC2229162", "primary_object": { "basename": "MCCjgp93.pdf", "url": "https://authors.library.caltech.edu/records/zn4rp-png14/files/MCCjgp93.pdf" }, "resource_type": "article", "pub_year": "1993", "author_list": "McCarthy, N. A.; McDonough, S.; et el." }, { "id": "https://authors.library.caltech.edu/records/8ff06-07074", "eprint_id": 63038, "eprint_status": "archive", "datestamp": "2023-08-22 09:10:03", "lastmod": "2023-10-25 23:16:50", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Uezono-Y", "name": { "family": "Uezono", "given": "Y." } }, { "id": "Bradley-J", "name": { "family": "Bradley", "given": "J." } }, { "id": "Min-C", "name": { "family": "Min", "given": "C." } }, { "id": "McCarty-N-A", "name": { "family": "McCarty", "given": "N. A." } }, { "id": "Quick-M-W", "name": { "family": "Quick", "given": "M." } }, { "id": "Riordan-J-R", "name": { "family": "Riordan", "given": "J. R." } }, { "id": "Chavkin-C", "name": { "family": "Chavkin", "given": "C." } }, { "id": "Zinn-K", "name": { "family": "Zinn", "given": "K." }, "orcid": "0000-0002-6706-5605" }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "N." } } ] }, "title": "Receptors that couple to 2 classes of G proteins increase cAMP and activate CFTR expressed in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "keywords": "Gs; Gi; cystic fibrosis; \u03b22 receptor; serotonin receptors", "note": "\u00a9 1993 Harwood Academic. \n\n(Received March 15, 1993; Revised May 15, 1993)\n\nWe thank Dr.s M. Simon and Anna Aragay for advice and the gift of Gs cDNA, R. Reed for the gift of the adenyl cyclase type II cDNA, and Dr. B. Kobilka for the gift of the \u03b22 receptor cDNA. We thank Drs. N. Dascal and C. Strader for comments on an earlier version of the manuscript. This work was supported by grants from the National Institutes of Health, the National Institute on Drug Abuse, the Silvio Conte Center for Neuroscience Research of the National Institute of Mental Health, the Cystic Fibrosis Foundation, the California Tobacco-Related Disease Program, the Wiersma Memorial Fund, and the Japanese Foundation for Clinical Pharmacology.", "abstract": "The cystic fibrosis transmembrane conductance regulator (CFTR), a Cl- channel activated by phosphorylation, was expressed in Xenopus oocytes along with various combinations of several other components of the cAMP signalling pathway. Activation of the coexpressed beta 2 adrenergic receptor increased cAMP and led to CFTR activation. The activation of CFTR (1) requires only short (15 s) exposure to isoproterenol, (2) occurs for agonist concentrations 100-1000 fold lower than those that produce cAMP increases detectable by a radioimmunoassay, (3) requires injection of only 5 pg of receptor cRNA per oocyte, and (4) can be increased further by coexpression of cRNA for adenylyl cyclase type II or III or for Gs alpha. In addition, CFTR activation and cAMP increases by beta 2 activation were enhanced by activation of the coexpressed 5HT1A receptor, which is thought to couple to Gi. The additional activation by the 5HT1A receptor was enhanced by coexpression of adenylyl cyclase type II but not with type III and may proceed via the beta gamma subunits of a G protein. The sensitivity of the assay system is also demonstrated by responses to vasoactive intestinal peptide and to pituitary adenylate cyclase-activating polypeptide in oocytes injected with cerebral cortex mRNA.", "date": "1993", "date_type": "published", "publication": "Receptors & Channels", "volume": "1", "number": "3", "publisher": "Harwood Academic", "pagerange": "233-241", "id_number": "CaltechAUTHORS:20151217-104726852", "issn": "1060-6823", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20151217-104726852", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "National Institute on Drug Abuse (NIDA)" }, { "agency": "National Institute of Mental Health (NIMH)" }, { "agency": "Cystic Fibrosis Foundation" }, { "agency": "California Tobacco-Related Disease Research Program" }, { "agency": "Wiersma Memorial Fund" }, { "agency": "Japanese Foundation for Clinical Pharmacology" } ] }, "resource_type": "article", "pub_year": "1993", "author_list": "Uezono, Y.; Bradley, J.; et el." }, { "id": "https://authors.library.caltech.edu/records/336nq-b5381", "eprint_id": 10848, "eprint_status": "archive", "datestamp": "2023-08-22 08:57:43", "lastmod": "2023-10-16 23:08:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Mutations in M2 alter the selectivity of the mouse nicotinic acetylcholine receptor for organic and alkali metal cations", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1992 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nOriginal version received 5 March 1992 and accepted version received 10 June 1992. \n\nWe thank Michael Quick and Linda Czyzyk for technical assistance. \n\nThis research was supported by grants from the National Institute of Health (NS-11756), the Muscular Dystrophy Association, and the University of California Tobacco-Related Disease Research Program.\n\nPublished - COHjgp92a.pdf
", "abstract": "We measured the permeability ratios (PX/PNa) of 3 wild-type, 1 hybrid, 2 subunit-deficient, and 22 mutant nicotinic receptors expressed in Xenopus oocytes for alkali metal and organic cations using shifts in the bi-ionic reversal potential of the macroscopic current. Mutations at three positions (2', 6', 10') in M2 affected ion selectivity. Mutations at position 2' (alpha Thr244, beta Gly255, gamma Thr253, delta Ser258) near the intracellular end of M2 changed the organic cation permeability ratios as much as twofold and reduced PCs/PNa and PK/PNa by 16-18%. Mutations at positions 6' and 10' increased the glycine ethyl ester/Na+ and glycine methyl ester/Na+ permeability ratios. Two subunit alterations also affected selectivity: omission of the delta subunit reduced PCs/PNa by 16%, and substitution of Xenopus delta for mouse delta increased Pguanidinium/PNa more than twofold and reduced PCs/PNa by 34% and PLi/PNa by 20%. The wild-type mouse receptor displayed a surprising interaction with the primary ammonium cations; relative permeability peaked at a chain length equal to four carbons. Analysis of the organic permeability ratios for the wild-type mouse receptor shows that (a) the diameter of the narrowest part of the pore is 8.4 A; (b) the mouse receptor departs significantly from size selectivity for monovalent organic cations; and (c) lowering the temperature reduces Pguanidinium/PNa by 38% and Pbutylammonium/PNa more than twofold. The results reinforce present views that positions -1' and 2' are the narrowest part of the pore and suggest that positions 6' and 10' align some permeant organic cations in the pore in an interaction similar to that with channel blocker, QX-222.", "date": "1992-09", "date_type": "published", "publication": "Journal of General Physiology", "volume": "100", "number": "3", "publisher": "Rockefeller University Press", "pagerange": "373-400", "id_number": "CaltechAUTHORS:COHjgp92a", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:COHjgp92a", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "California Tobacco-Related Disease Research Program" } ] }, "doi": "10.1085/jgp.100.3.373", "pmcid": "PMC2229089", "primary_object": { "basename": "COHjgp92a.pdf", "url": "https://authors.library.caltech.edu/records/336nq-b5381/files/COHjgp92a.pdf" }, "resource_type": "article", "pub_year": "1992", "author_list": "Cohen, Bruce N.; Labarca, Cesar; et el." }, { "id": "https://authors.library.caltech.edu/records/4f9gz-skc95", "eprint_id": 7441, "eprint_status": "archive", "datestamp": "2023-08-22 08:55:52", "lastmod": "2023-10-16 20:54:36", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Guastella-J", "name": { "family": "Guastella", "given": "John" } }, { "id": "Brecha-N", "name": { "family": "Brecha", "given": "Nicholas" } }, { "id": "Weigmann-C", "name": { "family": "Weigmann", "given": "Christine" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Cloning, expression, and localization of a rat brain high-affinity glycine transporter", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1992 by the National Academy of Sciences. \n\nContributed by Norman Davidson, May 6, 1992. \n\nWe thank M. Quick for oocyte preparation, L. Czyzyk for assistance with nucleotide sequencing, and A. Figl for assistance with oligonucleotide synthesis. This work was supported by National Institutes of Health Grant EY04067, Neuroendocrine Anatomy Core Grant DK4130, and Veterans Administration Medical Research Funds to N.B.; National Institutes of Health Grant NS-11756 to H.A.L. and N.D.; and a National Institutes of Health Postdoctoral Fellowship to J.G. \n\nThe sequence reported in this paper has been deposited in the GenBank data base (accession no. M95413). \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - GUApnas92.pdf
", "abstract": "A cDNA clone encoding a glycine transporter has been isolated from rat brain by a combined PCR and plaque-hybridization strategy. mRNA synthesized from this clone (designated GLYT1) directs the expression of sodium-and chloride-dependent, high-affinity uptake of [3H]glycine by Xenopus oocytes. [3H]Glycine transport mediated by clone GLYT1 is blocked by sarcosine but is not blocked by methylaminoisobutyric acid or L-alanine, a substrate specificity similar to that described for a previously identified glycine-uptake system called system Gly. In situ hybridization reveals that GLYT1 is prominently expressed in the cervical spinal cord and brainstem, two regions of the central nervous system where glycine is a putative neurotransmitter. GLYT1 is also strongly expressed in the cerebellum and olfactory bulb and is expressed at lower levels in other brain regions. The open reading frame of the GLYT1 cDNA predicts a protein containing 633 amino acids with a molecular mass of \u224870 kDa. The primary structure and hydropathicity profile of GLYT1 protein reveal that this protein is a member of the sodium- and chloride-dependent superfamily of transporters that utilize neurotransmitters and related substances as substrates.", "date": "1992-08-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "89", "number": "15", "publisher": "National Academy of Sciences", "pagerange": "7189-7193", "id_number": "CaltechAUTHORS:GUApnas92", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:GUApnas92", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "EY04067" }, { "agency": "NIH", "grant_number": "DK4130" }, { "agency": "Veterans Administration Medical Research Funds" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH Postdoctoral Fellowship" } ] }, "doi": "10.1073/pnas.89.15.7189", "pmcid": "PMC49671", "primary_object": { "basename": "GUApnas92.pdf", "url": "https://authors.library.caltech.edu/records/4f9gz-skc95/files/GUApnas92.pdf" }, "resource_type": "article", "pub_year": "1992", "author_list": "Guastella, John; Brecha, Nicholas; et el." }, { "id": "https://authors.library.caltech.edu/records/a65kf-d5n11", "eprint_id": 29896, "eprint_status": "archive", "datestamp": "2023-08-20 01:21:48", "lastmod": "2023-10-24 22:32:55", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "The Permeation Pathway of Neurotransmitter-Gated Ion Channels", "ispublished": "pub", "full_text_status": "restricted", "keywords": "synapse, nicotinic acetylcholine receptor, neurotransmitter\nreceptor, open-channel blockers, site-directed mutagenesis", "note": "\u00a9 1992 Annual Reviews.\n\nI thank B. Cohen for help with Figure 1 and Table 1, M. King for help\nwith the manuscript, and C. Jahr, N. Lim, N. McCarty, and D. Vaughn\nfor many helpful comments. N. Davidson has been my partner on all\nrecent studies and continues his vital contributions. Preparation of this\nreview was supported by grants from the Muscular Dystrophy Association,\nthe California Tobacco-Related Disease Research Program, and the\nNational Institutes of Health (NS-11756).", "abstract": "N/A", "date": "1992-06", "date_type": "published", "publication": "Annual Review of Biophysics and Biomolecular Structure", "volume": "21", "publisher": "Annual Reviews", "pagerange": "267-292", "id_number": "CaltechAUTHORS:20120329-091749403", "issn": "1056-8700", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120329-091749403", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "California Tobacco-Related Disease Research Program" } ] }, "doi": "10.1146/annurev.bb.21.060192.001411", "resource_type": "article", "pub_year": "1992", "author_list": "Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/2saaf-42t80", "eprint_id": 10847, "eprint_status": "archive", "datestamp": "2023-08-22 08:47:01", "lastmod": "2023-10-16 23:08:16", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Cohen-B-N", "name": { "family": "Cohen", "given": "Bruce N." } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Czyzyk-L", "name": { "family": "Czyzyk", "given": "Linda" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Tris+/Na+ permeability ratios of nicotinic acetylcholine receptors are reduced by mutations near the intracellular end of the M2 region", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1992 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nOriginal version received 19 July 1991 and accepted version received 13 December 1991. \n\nWe thank Euk Kwon and Michael Quick for technical assistance, Fred Sigworth and Gary Yellen for helpful discussion, and Pierre Charnet for much tutelage and helpful discussion. \n\nThis research was supported by grants from the National Institutes of Health (NS-11756) and the Muscular Dystrophy Association.\n\nPublished - COHjgp92b.pdf
", "abstract": "Tris+/Na+ permeability ratios were measured from shifts in the biionic reversal potentials of the macroscopic ACh-induced currents for 3 wild- type (WT), 1 hybrid, 2 subunit-deficient, and 25 mutant nicotinic receptors expressed in Xenopus oocytes. At two positions near the putative intracellular end of M2, 2' (alpha Thr244, beta Gly255, gamma Thr253, delta Ser258) and -1', point mutations reduced the relative Tris+ permeability of the mouse receptor as much as threefold. Comparable mutations at several other positions had no effects on relative Tris+ permeability. Mutations in delta had a greater effect on relative Tris+ permeability than did comparable mutations in gamma; omission of the mouse delta subunit (delta 0 receptor) or replacement of mouse delta with Xenopus delta dramatically reduced relative Tris+ permeability. The WT mouse muscle receptor (alpha beta gamma delta) had a higher relative permeability to Tris+ than the wild-type Torpedo receptor. Analysis of the data show that (a) changes in the Tris+/Na+ permeability ratio produced by mutations correlate better with the hydrophobicity of the amino acid residues in M2 than with their volume; and (b) the mole-fraction dependence of the reversal potential in mixed Na+/Tris+ solutions is approximately consistent with the Goldman- Hodgkin-Katz voltage equation. The results suggest that the main ion selectivity filter for large monovalent cations in the ACh receptor channel is the region delimited by positions -1' and 2' near the intracellular end of the M2 helix.", "date": "1992-04", "date_type": "published", "publication": "Journal of General Physiology", "volume": "99", "number": "4", "publisher": "Rockefeller University Press", "pagerange": "545-572", "id_number": "CaltechAUTHORS:COHjgp92b", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:COHjgp92b", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" } ] }, "doi": "10.1085/jgp.99.4.545", "pmcid": "PMC2219204", "primary_object": { "basename": "COHjgp92b.pdf", "url": "https://authors.library.caltech.edu/records/2saaf-42t80/files/COHjgp92b.pdf" }, "resource_type": "article", "pub_year": "1992", "author_list": "Cohen, Bruce N.; Labarca, Cesar; et el." }, { "id": "https://authors.library.caltech.edu/records/36rr0-zqx49", "eprint_id": 99633, "eprint_status": "archive", "datestamp": "2023-08-20 01:00:23", "lastmod": "2023-10-18 18:36:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Yang-Xian-cheng", "name": { "family": "Yang", "given": "Xian-cheng" } }, { "id": "Labarca-Cesar", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Nargeot-Joel", "name": { "family": "Nargeot", "given": "Joel" } }, { "id": "Ho-Begonia-Y", "name": { "family": "Ho", "given": "Begonia Y." } }, { "id": "Elroy-Stein-Orna", "name": { "family": "Elroy-Stein", "given": "Orna" } }, { "id": "Moss-Bernard", "name": { "family": "Moss", "given": "Bernard" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Cell-specific posttranslational events affect functional expression at the plasma membrane but not tetrodotoxin sensitivity of the rat brain IIA sodium channel \u03b1-subunit expressed in mammalian cells", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1992 Society for Neuroscience. \n\nReceived May 6, 1991; revised Aug. 15, 1991; accepted Aug. 26, 1991. \n\nWe thank Carolyn Nolan and Euk Kwon for excellent technical assistance. This work is supported by grants from the NIH (NS-11756 and GM-29836), the Muscular Dystrophy Association, the Multiple Sclerosis Society, and NATO (CRG 890374) and by postdoctoral fellowships from the Muscular Dystrophy Association\n(X.-C.Y.) and the American Heart Association, Greater Los Angeles Chapter (B.Y.H.).\n\nPublished - 268.full.pdf
", "abstract": "The rat brain IIA Na\u207a channel alpha-subunit was expressed and studied in mammalian cells. Cells were infected with a recombinant vaccinia virus (VV) carrying the bacteriophage T7 RNA polymerase gene and were transfected with cDNA encoding the IIA Na\u207a channel \u03b1-subunit under control of a T7 promoter. Whole-cell patch-clamp recording showed that functional IIA channels were expressed efficiently (~10 channels/ \u00b5m\u00b2 in approximately 60% of cells) in Chinese hamster ovary (CHO) cells and in neonatal rat ventricular myocytes but were expressed poorly in undifferentiated BC\u2083H1 cells and failed to express in Ltk\u207b cells. However, voltage-dependent Drosophila Shaker H4 K\u207a channels and Escherichia coli \u03b2-galactosidase were expressed efficiently in all four cell types with VV vectors. Because RNA synthesis probably occurs without major differences in the cytoplasm of all infected cell types under the control of the T7 promoter and T7 polymerase, we conclude that cell type-specific expression of the Na\u207a channel probably reflects differences at posttranslational steps. The gating properties of the IIA Na\u207a currents expressed in cardiac myocytes differed from those expressed in CHO cells; most noticeably, the IIA Na\u207a currents displayed more rapid macroscopic inactivation when expressed in cardiac myocytes. These differences also suggest cell- specific posttranslational modifications. IIA channels were blocked by ~90% by 90 nM TTX when expressed either in CHO cells or in cardiac myocytes; the latter also continued to display endogenous TTX- resistant Na\u207a currents. Therefore, the TTX binding site of the channel is not affected by cell-specific modifications and is encoded by the primary amino acid sequence.", "date": "1992-01-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "12", "number": "1", "publisher": "Society for Neuroscience", "pagerange": "268-277", "id_number": "CaltechAUTHORS:20191101-161058438", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191101-161058438", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Multiple Sclerosis Society" }, { "agency": "North Atlantic Treaty Organization (NATO)", "grant_number": "CRG 890374" }, { "agency": "American Heart Association, Greater Los Angeles Affiliate" } ] }, "doi": "10.1523/jneurosci.12-01-00268.1992", "pmcid": "PMC6575683", "primary_object": { "basename": "268.full.pdf", "url": "https://authors.library.caltech.edu/records/36rr0-zqx49/files/268.full.pdf" }, "resource_type": "article", "pub_year": "1992", "author_list": "Yang, Xian-cheng; Labarca, Cesar; et el." }, { "id": "https://authors.library.caltech.edu/records/0q58w-8xy93", "eprint_id": 9487, "eprint_status": "archive", "datestamp": "2023-08-22 08:25:18", "lastmod": "2023-10-23 19:33:04", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Strategies for studying permeation at voltage-gated ion channels", "ispublished": "pub", "full_text_status": "public", "keywords": "HETEROLOGOUS EXPRESSION; SODIUM POTASSIUM CALCIUM CHANNEL EXCITABLE MEMBRANES; LOCAL ANESTHETICS; SITE-DIRECTED MUTAGENESIS", "note": "\"Reprinted, with permission, from the Annual Review of Physiology, Volume 53 copyright 1991 by Annual Reviews, www.annualreviews.org\".\nI thank R. Aldrich and R. MacKinnon for sharing their work prior to publication, P. Lazarow for challenges on rectification, and N. Davidson, B. Hille, T. Begenisich, and members of my research group for many helpful comments. Preparation of this review was supported by grants from the Klingenstein Foundation, the Muscular Dystrophy Association, and the National Institutes of Health (GM-29836 and NS-11756).\n\nPublished - LESarp91.pdf
", "abstract": "Voltage-dependent ion channels are presently thought to consist of several distinct functional regions: (a) activation gates, (b) inactivation gates, and permeation pathways. This chapter focuses on permeation pathways and may spur new ideas about experiments that use site-directed mutagenesis to probe the ion conduction pathway. Some hubris is required to attempt a survey of this field since individual families -- K^+, Na^+, or Ca^(2+) -- have been reviewed in detail (15, 68, 115, 127). My unified treatment is motivated by the structural similarity suggested by recent cDNA sequencing data on this group (see, for instance, 24). There have been many excellent previous treatments of ion channel permeation (6, 15, 34, 35, 51, 53, 68, 73, 74, 115, 127).", "date": "1991-10", "date_type": "published", "publication": "Annual Review of Physiology", "volume": "53", "publisher": "Annual Reviews", "pagerange": "477-496", "id_number": "CaltechAUTHORS:LESarp91", "issn": "0066-4278", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LESarp91", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Klingenstein Foundation" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "primary_object": { "basename": "LESarp91.pdf", "url": "https://authors.library.caltech.edu/records/0q58w-8xy93/files/LESarp91.pdf" }, "resource_type": "article", "pub_year": "1991", "author_list": "Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/c035w-q0r76", "eprint_id": 30212, "eprint_status": "archive", "datestamp": "2023-08-20 00:17:00", "lastmod": "2023-10-17 15:33:20", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Walter-A-E", "name": { "family": "Walter", "given": "A. E." } }, { "id": "Hoger-J-H", "name": { "family": "Hoger", "given": "J. H." } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "C." } }, { "id": "Yu-L", "name": { "family": "Yu", "given": "L." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "N." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Low Molecular Weight mRNA Encodes a Protein That Controls Serotonin 5-HT_(1c) and Acetylcholine M_1 Receptor Sensitivity\n in Xenopus Oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1991 Rockefeller University Press. Original version received 9 October 1990 and accepted version received 2January 1991. We thank Dr. Tom I. Bonner for the rat M_1 receptor clone, Dr. N. Gautam for the G\u03b3 clones, and Annie Gouin and Euk Kwon for isolating the oocytes. This work was supported by fellowships from the American Heart Association to A. E. Walter and from the NIH and American Cancer Society to J. H. Hoger, and by grants from the NIH (NS-11756,\nGM-10991) and The Markey Trust.\n\nPublished - WALjgp91.pdf
", "abstract": "Serotonin 5-HT_(1c) and acetylcholine M_1 receptors activate phosphoinositidase, resulting in an increased formation of IP_3 and 1,2 diacylglycerol. In Xenopus oocytes injected with mRNA encoding either of these receptors, Ca^(2+) released from intracellular stores in response to IP3 then opens Ca^(2+)-gated Cl^-channels. In the present experiments, oocytes expressing a transcript from a cloned mouse serotonin 5-HT_(1c) receptor were exposed to identical 15-s pulses of agonist, administered 2 min apart; the second current response was two to three times that of the first. However, in those oocytes coinjected with the 5-HT_(1c) receptor transcript and a low molecular weight fraction (0.3-1.5 kb) of rat brain mRNA, the second current response was ~50% of the first. Thus, the low molecular weight RNA encodes a protein (or proteins) that causes desensitization. Experiments using fura-2 or a Ca^(2+)-free superfusate indicated that desensitization of the 5-HT_(1c) receptor response does not result from a sustained elevation of intracellular Ca^(2+) level or require the entry of extracellular Ca^(2+). Photolysis of caged IP_3 demonstrated that an increase in IP_3 and a subsequent rise in Ca^(2+) do not produce desensitization of either the IP_3 or 5-HT_(1c) peak current responses. Furthermore, in oocytes coinjected with the low molecular weight RNA and a transcript from the rat M_1 acetylcholine receptor, the M_1 current response was greatly attenuated. Our data suggest that the proteins involved in attenuation of the M_1 current response and desensitization of the 5-HT_(1c) current response may be the same.", "date": "1991-08", "date_type": "published", "publication": "Journal of General Physiology", "volume": "98", "number": "2", "publisher": "Rockefeller University Press", "pagerange": "399-417", "id_number": "CaltechAUTHORS:20120420-071637268", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120420-071637268", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "American Heart Association" }, { "agency": "American Cancer Society" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "Lucille P. Markey Charitable Trust" } ] }, "doi": "10.1085/jgp.98.2.399", "pmcid": "PMC2229050", "primary_object": { "basename": "WALjgp91.pdf", "url": "https://authors.library.caltech.edu/records/c035w-q0r76/files/WALjgp91.pdf" }, "resource_type": "article", "pub_year": "1991", "author_list": "Walter, A. E.; Hoger, J. H.; et el." }, { "id": "https://authors.library.caltech.edu/records/aj9vt-deh94", "eprint_id": 7540, "eprint_status": "archive", "datestamp": "2023-08-22 08:11:40", "lastmod": "2023-10-16 20:57:58", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Karschin-A", "name": { "family": "Karschin", "given": "Andreas" } }, { "id": "Ho-Begonia-Y", "name": { "family": "Ho", "given": "Begonia Y." } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Elroy-Stein-O", "name": { "family": "Elroy-Stein", "given": "Orna" } }, { "id": "Moss-B", "name": { "family": "Moss", "given": "Bernard" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Heterologously expressed serotonin 1A receptors couple to muscarinic K+ channels in heart", "ispublished": "pub", "full_text_status": "public", "keywords": "VACCINIA VIRUS EXPRESSION; GUANINE NUCLEOTIDE-BINDING PROTEINS; 7-HELIX RECEPTORS; ATRIAL CELLS; ACETYLCHOLINE", "note": "\u00a9 1991 by the National Academy of Sciences. \n\nContributed by Norman Davidson, April 2, 1991. \n\nWe thank R. Weinshank and P. Hartig for the human 5-HT1AR cDNA; T. Branchek and N. Adham for performing some binding experiments; and A. Gouin, E. Kwon, C. Nolan, and M. King for expert assistance. This work was supported by the National Institutes of Health (GM29836 and GM10991) and the Cystic Fibrosis Foundation and by fellowships from the Max Kade Foundation and Alexander von Humboldt Stiftung (A.K.) and the American Heart Association (B.Y.H.). \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - KARpnas91.pdf
", "abstract": "In cardiac atrial cells, muscarinic acetylcholine receptors activate a K+ current directly via a guanine nucleotide-binding protein (G protein). Serotonin type 1A receptors may activate a similar pathway in hippocampal neurons. To develop a system in which receptor/G protein/K+ channel coupling can be experimentally manipulated, we have used a highly efficient recombinant vaccinia virus vector system to express human serotonin 1A receptors in primary cultures of rat atrial myocytes. The expressed 1A receptors activated the inwardly rectifying K+ conductance that is normally activated by the endogenous muscarinic acetylcholine receptors. Maximal responses to either agonist occluded further activation by the other agonist. The average activation time constants for serotonin were about 5 times slower than for acetylcholine. The data support suggestions that the intracellular signaling pathway from seven-helix receptors to G proteins and directly to ion channels is widespread in excitable cells. After a fraction of the G proteins are activated irreversibly by guanosine 5'-[\u03b3-thio]triphosphate, subsequent transduction proceeds more efficiently. One possible interpretation is that multiple G-protein molecules are required to activate each channel. Vaccinia virus expression vectors are thus useful for expressing seven-helix receptors in primary cultures of postmitotic cells and have provided a heterologous expression system for the signaling pathway from seven-helix receptors to G proteins and directly to ion channels.", "date": "1991-07-15", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "88", "number": "13", "publisher": "National Academy of Sciences", "pagerange": "5694-5698", "id_number": "CaltechAUTHORS:KARpnas91", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:KARpnas91", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "GM10991" }, { "agency": "Cystic Fibrosis Foundation" }, { "agency": "Max Kade Foundation" }, { "agency": "Alexander von Humboldt Stiftung" }, { "agency": "American Heart Association" } ] }, "doi": "10.1073/pnas.88.13.5694", "pmcid": "PMC51944", "primary_object": { "basename": "KARpnas91.pdf", "url": "https://authors.library.caltech.edu/records/aj9vt-deh94/files/KARpnas91.pdf" }, "resource_type": "article", "pub_year": "1991", "author_list": "Karschin, Andreas; Ho, Begonia Y.; et el." }, { "id": "https://authors.library.caltech.edu/records/cj9pd-z5t88", "eprint_id": 13228, "eprint_status": "archive", "datestamp": "2023-08-19 23:11:15", "lastmod": "2023-10-17 22:46:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Ahmed-C-M-I", "name": { "family": "Ahmed", "given": "C. M. lqbal" } }, { "id": "Auld-V-J", "name": { "family": "Auld", "given": "Vanessa J." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dunn-R-J", "name": { "family": "Dunn", "given": "Robert" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Both sodium channel II and IIA alpha subunits are expressed in rat brain", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1990 Oxford University Press. \n\nSubmitted August 31, 1990.\n\nPublished - AHMnar90.pdf
", "abstract": "Complementary DNAs encoding four distinct isoforms of the large (\u03b1) subunit of the voltage sensitive sodium channel have been isolated from rat brain mRNA (1,2,3). Two of these, II and HA, differ in only 36 nucleotides within the coding region and in only 6 amino acids (1, 3). Because of the possible functional significance of electrophysiological differences between isoforms, it is important to establish definitively that the sequence differences are not due to cloning artifacts and to estimate the relative prevalence levels of the two mRNAs in rat brain.", "date": "1990-10-11", "date_type": "published", "publication": "Nucleic Acids Research", "volume": "18", "number": "19", "publisher": "Oxford University Press", "pagerange": "5907-5907", "id_number": "CaltechAUTHORS:AHMnar90", "issn": "0305-1048", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:AHMnar90", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1093/nar/18.19.5907", "pmcid": "PMC332354", "primary_object": { "basename": "AHMnar90.pdf", "url": "https://authors.library.caltech.edu/records/cj9pd-z5t88/files/AHMnar90.pdf" }, "resource_type": "article", "pub_year": "1990", "author_list": "Ahmed, C. M. lqbal; Auld, Vanessa J.; et el." }, { "id": "https://authors.library.caltech.edu/records/phcjx-xks66", "eprint_id": 5447, "eprint_status": "archive", "datestamp": "2023-08-22 07:45:42", "lastmod": "2023-10-16 19:13:31", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Krafte-D-S", "name": { "family": "Krafte", "given": "Douglas S." } }, { "id": "Goldin-A-L", "name": { "family": "Goldin", "given": "Alan L." } }, { "id": "Auld-V-J", "name": { "family": "Auld", "given": "Vanessa J." } }, { "id": "Dunn-R-J", "name": { "family": "Dunn", "given": "Robert J." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Inactivation of cloned Na channels expressed in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1990 by The Rockefeller University Press. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. \n\nOriginal version received 10 October 1989 and accepted version received 27 April 1990. \n\nWe thank A. Gouin and W. Conley for excellent technical assistance, Dr. R.J. Leonard and Dr. I. Ahmed for helpful suggestions during the course of this work, and Dr. W. A. Catterall for comments on the manuscript. \n\nThis research was supported by grants from the U.S. National Institutes of Health (NS-11756, GM-10991, and NS-26729), the Lucille P. Markey Charitable Trust, the Esther A. and Joseph Klingenstein Fund, the Multiple Sclerosis Society (Canada), the Medical Research Council (Canada), and the March of Dimes Basil O'Connor Starter Scholar Program. D.S. Krafte held a postdoctoral fellowship from the NIH. A.L. Goldin is a Lucille P. Markey Scholar.\n\nPublished - KRAjgp90.pdf
", "abstract": "This study investigates the inactivation properties of Na channels expressed in Xenopus oocytes from two rat IIA Na channel cDNA clones differing by a single amino acid residue. Although the two cDNAs encode Na channels with substantially different activation properties (Auld, V. J., A. L. Goldin, D. S. Krafte, J. Marshall, J. M. Dunn, W. A. Catterall, H. A. Lester, N. Davidson, and R. J. Dunn. 1988. Neuron. 1:449-461), their inactivation properties resemble each other strongly but differ markedly from channels induced by poly(A+) rat brain RNA. Rat IIA currents inactivate more slowly, recover from inactivation more slowly, and display a steady-state voltage dependence that is shifted to more positive potentials. The macroscopic inactivation process for poly(A+) Na channels is defined by a single exponential time course; that for rat IIA channels displays two exponential components. At the single-channel level these differences in inactivation occur because rat IIA channels reopen several times during a depolarizing pulse; poly(A+) channels do not. Repetitive stimulation (greater than 1 Hz) produces a marked decrement in the rat IIA peak current and changes the waveform of the currents. When low molecular weight RNA is coinjected with rat IIA RNA, these inactivation properties are restored to those that characterize poly(A+) channels. Slow inactivation is similar for rat IIA and poly(A+) channels, however. The data suggest that activation and inactivation involve at least partially distinct regions of the channel protein.", "date": "1990-10", "date_type": "published", "publication": "Journal of General Physiology", "volume": "96", "number": "4", "publisher": "Rockefeller University Press", "pagerange": "689-706", "id_number": "CaltechAUTHORS:KRAjgp90", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:KRAjgp90", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "NS-26729" }, { "agency": "Lucille P. Markey Charitable Trust" }, { "agency": "Esther A. and Joseph Klingenstein Fund" }, { "agency": "Multiple Sclerosis Society of Canada" }, { "agency": "Medical Research Council of Canada" }, { "agency": "March of Dimes Foundation" }, { "agency": "NIH Postdoctoral Fellowship" } ] }, "doi": "10.1085/jgp.96.4.689", "pmcid": "PMC2229013", "primary_object": { "basename": "KRAjgp90.pdf", "url": "https://authors.library.caltech.edu/records/phcjx-xks66/files/KRAjgp90.pdf" }, "resource_type": "article", "pub_year": "1990", "author_list": "Krafte, Douglas S.; Goldin, Alan L.; et el." }, { "id": "https://authors.library.caltech.edu/records/smwpv-b5x55", "eprint_id": 53114, "eprint_status": "archive", "datestamp": "2023-08-19 23:06:13", "lastmod": "2023-10-19 14:33:22", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Guastella-John", "name": { "family": "Guastella", "given": "John" } }, { "id": "Nelson-Nathan", "name": { "family": "Nelson", "given": "Nathan" } }, { "id": "Nelson-Hannah", "name": { "family": "Nelson", "given": "Hannah" } }, { "id": "Czyzyk-Linda", "name": { "family": "Czyzyk", "given": "Linda" } }, { "id": "Keynan-Shoshi", "name": { "family": "Keynan", "given": "Shoshi" } }, { "id": "Miedel-May-C", "name": { "family": "Miedel", "given": "May C." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Kanner-Baruch", "name": { "family": "Kanner", "given": "Baruch" } } ] }, "title": "Cloning and expression of a rat brain GABA transporter", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1990 American Association for the Advancement of Science.\n\n30 March 1990; Accepted 8 June 1990.\n\nWe thank R. Radian and A. Bendahan for protein purification, and I. Ahmed, J. Hoger, D. Krafte, C. LaBarca, T. Snutch, and A. Walter for sharing protocols and expertise. We also thank T. Snutch for participating in the construction of the cDNA library, S. Celniker and the Caltech fly group for the use of their oligonucleotide synthesizer, and S. Bajjalieh for comments on the manuscript. We especially thank A. Gouin for oocyte preparation. Supported by NIH grants GM 29836, GM 10991,\nand NS 16708, by U.S.-Israel Binational Science Foundation grant 86-00147, and by an NIH postdoctoral fellowship to J.G.", "abstract": "A complementary DNA clone (designated GAT-1) encoding a transporter for the neurotransmitter gamma-aminobutyric acid (GABA) has been isolated from rat brain, and its functional properties have been examined in Xenopus oocytes. Oocytes injected with GAT-1 synthetic messenger RNA accumulated [^3H]GABA to levels above control values. The transporter encoded by GAT-1 has a high affinity for GABA, is sodium-and chloride-dependent, and is pharmacologically similar to neuronal GABA transporters. The GAT-1 protein shares antigenic determinants with a native rat brain GABA transporter. The nucleotide sequence of GAT-1 predicts a protein of 599 amino acids with a molecular weight of 67 kilodaltons. Hydropathy analysis of the deduced protein suggests multiple transmembrane regions, a feature shared by several cloned transporters; however, database searches indicate that GAT-1 is not homologous to any previously identified proteins. Therefore, GAT-1 appears to be a member of a previously uncharacterized family of transport molecules.", "date": "1990-09-14", "date_type": "published", "publication": "Science", "volume": "249", "number": "4974", "publisher": "American Association for the Advancement of Science", "pagerange": "1303-1306", "id_number": "CaltechAUTHORS:20141222-150126708", "issn": "0036-8075", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20141222-150126708", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM 29836" }, { "agency": "NIH", "grant_number": "GM 10991" }, { "agency": "NIH", "grant_number": "NS 16708" }, { "agency": "Binational Science Foundation (USA-Israel)", "grant_number": "86-00147" }, { "agency": "NIH Postdoctoral Fellowship" } ] }, "doi": "10.1126/science.1975955", "resource_type": "article", "pub_year": "1990", "author_list": "Guastella, John; Nelson, Nathan; et el." }, { "id": "https://authors.library.caltech.edu/records/j4ybj-20829", "eprint_id": 8754, "eprint_status": "archive", "datestamp": "2023-08-22 07:36:48", "lastmod": "2023-10-16 21:41:56", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "Leonard-J-P", "name": { "family": "Leonard", "given": "John P." } }, { "id": "Gilbert-M-M", "name": { "family": "Gilbert", "given": "Mary M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Rat brain expresses a heterogeneous family of calcium channels", "ispublished": "pub", "full_text_status": "public", "keywords": "cDNA cloning; hybrid arrest; dihydropyridine receptor", "note": "\u00a9 1990 by the National Academy of Sciences. \n\nContributed by Norman Davidson, February 21, 1990. \n\nWe are grateful to Steve Dubel for technical assistance and to Ellis et al. (11) for providing the rabbit Ca-channel probe. We also thank Mike White for the suggestion of using niflumic acid to block the oocyte C1 current. This work was supported by grants from the Medical Research Council of Canada (T.P.S.) and National Institutes of Health Grants NS-26432 (J.P.L.), GM-10991 (N.D.), and GM-29836 (H.A.L.). During the earlier phases of this work, T.P.S. was supported by an American Heart Association Fellowship, Greater Los Angeles Affiliate, at the California Institute of Technology. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - SNUpnas90.pdf
", "abstract": "We describe the isolation and characterization of several rat brain cDNAs that are homologous to the 1 subunit of heart and skeletal muscle dihydropyridine-sensitive Ca channels. Northern blot analysis of 32 cDNAs shows that they can be grouped into four distinct classes (A, B, C, and D), each corresponding to a distinct hybridization pattern of brain mRNAs. Southern blot and DNA sequencing suggest that each class of cDNA represents a distinct gene or gene family. In the regions sequenced, the rat brain class C and D gene products share 75% amino acid identity with the rabbit skeletal muscle Ca channel. In addition, the class C polypeptide is almost identical to the rabbit cardiac Ca channel (97% identity). In contrast, the rat brain class A and B cDNAs are more distantly related to dihydropyridine-sensitive Ca channels (47-64% amino acid identity) and to the brain class C and D genes (51-55% amino acid identity). To examine the functional significance of the isolated brain cDNAs, hybrid depletion experiments were performed in Xenopus oocytes. Antisense oligonucleotides against class A and B cDNAs each partially inhibited, and a class C oligonucleotide almost fully inhibited, the expression of Ba current in rat brain mRNA injected oocytes; but none of the oligonucleotides affected the expression of voltage-gated Na or K conductances. The clone characterization and sequencing results demonstrate that a number of distinct, yet related, voltage-gated Ca-channel genes are expressed in the brain. The antisense oligonucleotide experiments specifically show that one or several of the Ca-channel classes are related to the Ca channels observed in rat brain mRNA injected oocytes.", "date": "1990-05-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "87", "number": "9", "publisher": "National Academy of Sciences", "pagerange": "3391-3395", "id_number": "CaltechAUTHORS:SNUpnas90", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:SNUpnas90", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Medical Research Council of Canada" }, { "agency": "NIH", "grant_number": "NS-26432" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "American Heart Association, Greater Los Angeles Affiliate" } ] }, "doi": "10.1073/pnas.87.9.3391", "pmcid": "PMC53906", "primary_object": { "basename": "SNUpnas90.pdf", "url": "https://authors.library.caltech.edu/records/j4ybj-20829/files/SNUpnas90.pdf" }, "resource_type": "article", "pub_year": "1990", "author_list": "Snutch, Terry P.; Leonard, John P.; et el." }, { "id": "https://authors.library.caltech.edu/records/56545-4z153", "eprint_id": 6962, "eprint_status": "archive", "datestamp": "2023-08-22 07:06:54", "lastmod": "2023-10-23 17:22:57", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Auld-V-J", "name": { "family": "Auld", "given": "V. J." } }, { "id": "Goldin-A-L", "name": { "family": "Goldin", "given": "A. L." } }, { "id": "Krafte-D-S", "name": { "family": "Krafte", "given": "D. S." } }, { "id": "Catterall-W-A", "name": { "family": "Catterall", "given": "W. A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "N." } }, { "id": "Dunn-R-J", "name": { "family": "Dunn", "given": "R. J." } } ] }, "title": "A Neutral Amino Acid Change in Segment IIS4 Dramatically Alters the Gating Properties of the Voltage-Dependent Sodium Channel", "ispublished": "pub", "full_text_status": "public", "keywords": "ion channel; site-directed mutagenesis; voltage clamp; Xenopus oocyte", "note": "\u00a9 1990 by National Academy of Sciences \n\nContributed by N. Davidson, September 21, 1989 \n\nWe thank K. McCormack and co-workers for communication of their observations prior to publication and R. Sarao for technical assistance. This research has been supported by grants from the National Institutes of Health (to N.D., H.A.L., A.L.G., and W.A.C.), by a Klingenstein Foundation grant (to H.A.L.), and by grants from the Multiple Sclerosis Society of Canada and the Medical Research Council of Canada (to R.J.D.). A.L.G. is a Lucille P. Markey Scholar and this work was also supported in part by a grant from the Lucille P. Markey Charitable Trust. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - AULpnas90.pdf
", "abstract": "Sodium channels encoded by the rat IIA cDNA clone [Auld, V. J., Goldin, A. L., Krafte, D. S., Marshall, J., Dunn, J., Catterall, W. A., Lester, H. A., Davidson, N. & Dunn, R. J. (1988) Neuron 1, 449-461] differ at seven amino acid residues from those encoded by the rat II cDNA [Noda, M., Ikeda, T., Kayano, T., Suzuki, H., Takeshima, H., Kurasaki, M., Takahashi, H. & Numa, S. (1986) Nature (London) 320, 188-192]. When expressed in Xenopus oocytes, rat IIA channels display a current-voltage relationship that is shifted 20-25 mV in the depolarizing direction relative to channels expressed from rat II cDNA or rat brain poly(A)+ mRNA. By modifying each variant residue in rat IIA to the corresponding residue in rat II, we demonstrate that a single Phe \u2192 Leu substitution at position 860 in the S4 segment of domain II is sufficient to shift the current-voltage relationship to that observed for channels expressed from rat brain poly(A)+ RNA or rat II cDNA. Rat genomic DNA encodes leucine but not phenylalanine at position 860, indicating that the phenylalanine at this position in rat IIA cDNA likely results from reverse transcriptase error.", "date": "1990-01-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "87", "number": "1", "publisher": "National Academy of Sciences", "pagerange": "323-327", "id_number": "CaltechAUTHORS:AULpnas90", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:AULpnas90", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "Esther A. and Joseph Klingenstein Fund" }, { "agency": "Multiple Sclerosis Society of Canada" }, { "agency": "Medical Research Council of Canada" }, { "agency": "Lucille P. Markey Charitable Trust" } ] }, "pmcid": "PMC53255", "primary_object": { "basename": "AULpnas90.pdf", "url": "https://authors.library.caltech.edu/records/56545-4z153/files/AULpnas90.pdf" }, "resource_type": "article", "pub_year": "1990", "author_list": "Auld, V. J.; Goldin, A. L.; et el." }, { "id": "https://authors.library.caltech.edu/records/ty7c7-wam06", "eprint_id": 76144, "eprint_status": "archive", "datestamp": "2023-08-19 22:21:27", "lastmod": "2023-10-25 15:32:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Charnet-P", "name": { "family": "Charnet", "given": "Pierre" } }, { "id": "Labarca-C", "name": { "family": "Labarca", "given": "Cesar" } }, { "id": "Leonard-R-J", "name": { "family": "Leonard", "given": "Reid J." } }, { "id": "Vogelaar-N-J", "name": { "family": "Vogelaar", "given": "Nancy J." } }, { "id": "Czyzyk-L", "name": { "family": "Czyzyk", "given": "Linda" } }, { "id": "Gouin-A", "name": { "family": "Gouin", "given": "Annie" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "An open-channel blocker interacts with adjacent turns of \u03b1-helices in the nicotinic acetylcholine receptor", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1990 Cell Press. \n\nReceived 17 August 1989, Revised 29 August 1989. \n\nWe thank Dr. Bertil Takmann of Astra Pharmaceuticals for gifts of QX-222; Drs. Takmann and R. A. North for discussion; and W. J. Goddard for the use of computer modeling facilities. This work was supported by grants from the National Institutes of Health (NS-11756) and from the Muscular Dystrophy Association and by postdoctoral fellowships to R. J. L. (NS-8083) and P. C. (Bourse Lavoiser and Fondation pour la Recherche M\u00e9dicale).", "abstract": "The binding site for an open-channel blocker, QX-222, at mouse muscle nicotinic acetylcholine receptors was probed using site-directed mutagenesis, oocyte expression, and elect rophysiologicaI analysis. The proposed cytoplasmic end of the M2 transmembrane helix is termed position 1\u2032. At position 10\u2032 (\u03b1S252, \u03b2T263, \u03b3A261, \u03b4A266), Ala residues yield stronger and longer binding of QX-222 than Ser or Thr residues. These effects are opposite and roughly equal (30%\u201350% per mutation) to previously reported effects at position 6\u2032. The polar end of an anesthetic molecule seems to bind to the position 6\u2032 OH groups, which provide a water-like region; the nonpolar moiety is near position 10\u2032 and binds more strongly in a nonpolar environment. Interactions with adjacent OH-rich turns of an amphiphilic helix may explain the widespread blocking effects of local anesthetics at the conduction pore of ion channels.", "date": "1990-01", "date_type": "published", "publication": "Neuron", "volume": "4", "number": "1", "publisher": "Neuron", "pagerange": "87-95", "id_number": "CaltechAUTHORS:20170408-161145019", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20170408-161145019", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-8083" }, { "agency": "Bourse Lavoiser" }, { "agency": "Fondation pour la Recherche M\u00e9dicale" } ] }, "doi": "10.1016/0896-6273(90)90445-L", "resource_type": "article", "pub_year": "1990", "author_list": "Charnet, Pierre; Labarca, Cesar; et el." }, { "id": "https://authors.library.caltech.edu/records/6xmv6-sdg88", "eprint_id": 12702, "eprint_status": "archive", "datestamp": "2023-08-19 22:05:28", "lastmod": "2023-10-17 20:26:44", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Yu-Lei", "name": { "family": "Yu", "given": "Lei" } }, { "id": "Blumer-K-J", "name": { "family": "Blumer", "given": "Kendall J." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Thorner-J", "name": { "family": "Thorner", "given": "Jeremy" } } ] }, "title": "Functional expression of the yeast alpha-factor receptor in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1989 by the American Society for Biochemistry and Molecular Biology. \n\n(Received for publication, June 19, 1989) \n\nWe thank Susan Porter for the isolation of the yeast GPA1 gene. \n\n\nThis work was supported by Grants NS11756, GM10991, and GM21841 from the National Institutes of Health. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. \n\n[L.Y. was] [s]upported by a Del Webb postdoctoral fellowship. \n\n[K.J.M. was] [s]upported by American Cancer Society Postdoctoral Fellowship PF2632.\n\nPublished - YULjbc89.pdf
", "abstract": "The STE2 gene of the yeast Saccharomyces cerevisiae encodes a 431- residue polypeptide that has been shown by chemical cross-linking and genetic studies to be a component of the receptor for the peptide mating pheromone, alpha-factor. To demonstrate directly that the ligand binding site of the alpha-factor receptor is comprised solely of the STE2 gene product, the STE2 protein was expressed in Xenopus oocytes. Oocytes microinjected with synthetic STE2 mRNA displayed specific surface binding for 35S-labeled alpha-factor (up to 40 sites/micron2/ng RNA). Oocytes injected with either STE2 antisense RNA or heterologous receptor mRNA (nicotinic acetylcholine receptor alpha, beta, gamma, and delta subunit mRNAs) showed no binding activity (indistinguishable from uninjected control oocytes). The apparent KD (7 nM) of the alpha-factor binding sites expressed on the oocyte surface, determined by competition binding studies, agreed with the values reported for intact yeast cells and yeast plasma membrane fractions. These findings demonstrate that the STE2 gene product is the only yeast polypeptide required for biogenesis of a functional alpha-factor receptor. Electrophysiological measurements indicated that the membrane conductance of oocytes injected with STE2 mRNA, or with both STE2 and GPA1 (encoding a yeast G protein alpha-subunit) mRNAs, did not change and was not affected by pheromone binding. Thus, the alpha-factor receptor, like mammalian G protein-coupled receptors, apparently lacks activity as an intrinsic or ligand-gated ion channel. This report is the first instance in which a membrane-bound receptor from a unicellular eukaryote has been expressed in a vertebrate cell.", "date": "1989-12-15", "date_type": "published", "publication": "Journal of Biological Chemistry", "volume": "264", "number": "35", "publisher": "American Society for Biochemistry and Molecular Biology", "pagerange": "20847-20850", "id_number": "CaltechAUTHORS:YULjbc89", "issn": "0021-9258", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:YULjbc89", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "NIH", "grant_number": "GM10991" }, { "agency": "NIH", "grant_number": "GM21841" }, { "agency": "Del E. Webb Foundation" }, { "agency": "American Cancer Society", "grant_number": "PF2632" } ] }, "primary_object": { "basename": "YULjbc89.pdf", "url": "https://authors.library.caltech.edu/records/6xmv6-sdg88/files/YULjbc89.pdf" }, "resource_type": "article", "pub_year": "1989", "author_list": "Yu, Lei; Blumer, Kendall J.; et el." }, { "id": "https://authors.library.caltech.edu/records/e7ab1-ypy93", "eprint_id": 8918, "eprint_status": "archive", "datestamp": "2023-08-22 06:53:54", "lastmod": "2023-10-23 17:23:00", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Leonard-R-J", "name": { "family": "Leonard", "given": "R. J." } }, { "id": "Karschin-A", "name": { "family": "Karschin", "given": "A." } }, { "id": "Jayashree-Aiyar-S", "name": { "family": "Jayashree-Aiyar", "given": "S." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "N." } }, { "id": "Tanouye-M-A", "name": { "family": "Tanouye", "given": "M. A." } }, { "id": "Thomas-L", "name": { "family": "Thomas", "given": "L." } }, { "id": "Thomas-G", "name": { "family": "Thomas", "given": "G." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Expression of Drosophila Shaker potassium channels in mammalian cells infected with recombinant vaccinia virus", "ispublished": "pub", "full_text_status": "public", "keywords": "heterologous expression; rat basophilic leukemia cells; rat pheochromocytoma cells", "note": "\u00a9 1989 by the National Academy of Sciences. \n\nContributed by N. Davidson, July 14, 1989. \n\nWe thank Barbara Thorne for expert assistance in the construction of the recombinant virus and J.H. Strauss and R.W. Aldrich for advice. Charybdotoxin was a generous gift of Dr. M.L. Garcia (Merck Sharp & Dohme Research Laboratories). This work was supported by National Institutes of Health Grants GM10991, GM29836, and DK37274, by the Cystic Fibrosis Foundation, and by fellowships from the Max Kade Foundation (A.K.) and the National Institutes of Health (R.J.L.). \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - LEOpnas89.pdf
", "abstract": "A recombinant vaccinia virus containing a Drosophila potassium channel (Shaker H4) cDNA was constructed by homologous recombination between wild-type vaccinia virus DNA and a transfer plasmid. The new virus was used to infect four types of mammalian cells in culture. Electrophysiological recording 24-72 hr after infection revealed the expression of voltage-gated transient potassium channels in all four cell types. The properties of the induced currents were identical to those previously observed following injection of the Shaker H4 transcript into oocytes. Vaccinia promises to be an effective vehicle for the heterologous expression of transmembrane ion channels in a variety of cell types.", "date": "1989-10-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "86", "number": "19", "publisher": "National Academy of Sciences", "pagerange": "7629-7633", "id_number": "CaltechAUTHORS:LEOpnas89", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LEOpnas89", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM10991" }, { "agency": "NIH", "grant_number": "GM29836" }, { "agency": "NIH", "grant_number": "DK37274" }, { "agency": "Cystic Fibrosis Foundation" }, { "agency": "Max Kade Foundation" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "pmcid": "PMC298120", "primary_object": { "basename": "LEOpnas89.pdf", "url": "https://authors.library.caltech.edu/records/e7ab1-ypy93/files/LEOpnas89.pdf" }, "resource_type": "article", "pub_year": "1989", "author_list": "Leonard, R. J.; Karschin, A.; et el." }, { "id": "https://authors.library.caltech.edu/records/rtq0q-a3h65", "eprint_id": 106003, "eprint_status": "archive", "datestamp": "2023-08-19 21:42:48", "lastmod": "2023-10-20 22:58:44", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "Leonard-J-P", "name": { "family": "Leonard", "given": "John P." } }, { "id": "Nargeot-J", "name": { "family": "Nargeot", "given": "Joel" } }, { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Curtis-B-M", "name": { "family": "Curtis", "given": "Benson M." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Expression of mRNA Encoding Voltage-Dependent Ca Channels in Xenopus Oocytes: Review and Progress Report", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1989 New York Academy of Sciences. \n\nThis research was sponsored by grants from the National Institutes of Health (GM\u201010991 and GM\u201029836), from the United States\u2014Israel Binational Science Foundation, from the CNRS (LP 8402), and from INSERM (U249), by postdoctoral fellowships from the American Heart Association to T. S. and J. P. L., from the NIH to B. M. C., and by a Bantrell Fellowship to N. D. J. N. thanks Laboratoire Servier for a travel grant.", "abstract": "We are studying electrically excitable Ca channels with an approach that combines nucleic acid molecular biology and membrane biophysics. Gurdon et al. first employed Xenopus oocytes as a system for heterologous expression of RNA. This system was later adapted for electrophysiological studies of ion channels by Barnard, Miledi, and their colleagues. Although other expression systems are being developed for membrane channels [Claudio et al. and Beam et al., this volume], the oocyte is now the most common, robust, and best-characterized system. Experiments in which channels are expressed by RNA injections in oocytes have at least three major goals.", "date": "1989-06", "date_type": "published", "publication": "Annals of the New York Academy of Sciences", "volume": "560", "publisher": "New York Academy of Sciences", "pagerange": "174-182", "id_number": "CaltechAUTHORS:20201012-160322461", "issn": "0077-8923", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-160322461", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "Centre National de la Recherche Scientifique (CNRS)", "grant_number": "LP 8402" }, { "agency": "Institut national de la sant\u00e9 et de la recherche m\u00e9dicale (INSERM)", "grant_number": "U249" }, { "agency": "American Heart Association" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "Laboratoire Servier" } ] }, "doi": "10.1111/j.1749-6632.1989.tb24094.x", "resource_type": "article", "pub_year": "1989", "author_list": "Lester, Henry A.; Snutch, Terry P.; et el." }, { "id": "https://authors.library.caltech.edu/records/q3mxn-cgn77", "eprint_id": 106002, "eprint_status": "archive", "datestamp": "2023-08-22 06:12:49", "lastmod": "2023-10-20 22:58:35", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Krafte-D-S", "name": { "family": "Krafte", "given": "Douglas S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Expression of functional sodium channels in stage II\u2013III Xenopus oocytes", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Xenopus oocyte; Na channel; Inactivation; mRNA; Expression system", "note": "\u00a9 1989 Published by Elsevier. \n\nReceived 20 April 1988, Revised 8 July 1988, Accepted 13 July 1988. \n\nWe wish to thank Norman Davidson for helpful\ndiscussion. This work was supported by NS-11756, GM-10991, and an NRSA fellowship to D.S.K.", "abstract": "We have injected mRNA from rabbit brain into stage II\u2013III Xenopus oocytes to determine whether they will translate exogenous RNA and incorporate functional ion channels into the membrane. Our results show that 48 h after RNA injection, functional voltage-dependent Na channels are present at sufficient densities to allow quantitative electrophysiological recording. The smaller oocytes have at least 2 experimental advantages over the stage V\u2013VI oocytes normally used for electrophysiological experiments: (1) the smaller membrane capacitance (approximately 5-fold) allows a faster settling time following a voltage step and a more detailed kinetic analysis of membrane currents than was previously possible with the 2-microelectrode technique, and (2) roughly 8-fold less RNA is needed for each injection. Thus, the stage II\u2013III Xenopus oocyte is a suitable preparation for the study of ion channels.", "date": "1989-01", "date_type": "published", "publication": "Journal of Neuroscience Methods", "volume": "26", "number": "3", "publisher": "Elsevier", "pagerange": "211-215", "id_number": "CaltechAUTHORS:20201012-160322367", "issn": "0165-0270", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-160322367", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "National Research Service Award" } ] }, "doi": "10.1016/0165-0270(89)90118-0", "resource_type": "article", "pub_year": "1989", "author_list": "Krafte, Douglas S. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/dpyaz-xy981", "eprint_id": 105980, "eprint_status": "archive", "datestamp": "2023-08-22 06:12:33", "lastmod": "2023-10-20 22:56:41", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Fong-Tung-Ming", "name": { "family": "Fong", "given": "Tung Ming" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Further evidence demonstrating that N-methyl-D-aspartate and kainate activate distinct ion channels", "ispublished": "pub", "full_text_status": "public", "keywords": "Excitatory amino acid; Rat brain; Xenopus oocytes; Open channel blocker; Competitive antagonists", "note": "\u00a9 1989 Alan R. Liss, Inc. \n\nManuscript accepted: 12 March 1989; Manuscript received: 11 January 1989.", "abstract": "Several excitatory amino acid receptors encoded by rat brain mRNA were expressed in Xenopus oocytes. Experimental protocols using an open channel blocker (MK\u2010801) were designed to test the common receptor\u2010channel hypothesis in which N\u2010methyl\u2010D\u2010aspartate (NMDA) and kainate activate the same ion channel but induce different open channel conformations with different ionic permeabilities. The present data demonstrate that NMDA exposes previously trapped MK\u2010801 molecules to the transmembrane field and accelerates their dissociation from the channel at positive potentials, while kainate lacks this effect, Therefore, kainate does not activate the same ion channel as NMDA does. Furthermore, differential inhibition of the NMDA response or the kainate response by the competitive antagonists D\u20102\u2010amino\u20105\u2010phosphonopentanoic acid (D\u2010AP5) and 6\u2010cyano\u20107\u2010nitroquinoxaline\u20102,3\u2010dione (CNQX) indicates that NMDA and kainate do not share the same binding site. Thus, these several lines of evidence demonstrate that two distinct receptor\u2010channels are activated by NMDA and kainate, respectively.", "date": "1989", "date_type": "published", "publication": "Synapse", "volume": "4", "number": "1", "publisher": "Wiley", "pagerange": "88-95", "id_number": "CaltechAUTHORS:20201012-122452479", "issn": "0887-4476", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-122452479", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1002/syn.890040110", "resource_type": "article", "pub_year": "1989", "author_list": "Fong, Tung Ming; Davidson, Norman; et el." }, { "id": "https://authors.library.caltech.edu/records/bf4rv-ss269", "eprint_id": 54199, "eprint_status": "archive", "datestamp": "2023-08-19 21:05:46", "lastmod": "2023-10-19 23:45:21", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Leonard-R-J", "name": { "family": "Leonard", "given": "Reid J." } }, { "id": "Labarca-C-G", "name": { "family": "Labarca", "given": "Cesar G." } }, { "id": "Charnet-P", "name": { "family": "Charnet", "given": "Pierre" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Evidence that the M2 membrane-spanning region lines the ion channel pore of the nicotinic receptor", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1988 American Association for the Advancement of Science.\n\n30 June 1988; Accepted 11 October 1988.\n\nWe thank M. Fearey and A. Charnet for help with the oocytes, L. Czyzyk for expert technical help with the mutagenesis, and G. Eisenman for discussion. This research was supported by grants from NIH (NS-11756) and the Muscular Dystrophy Association of America and by postdoctoral fellowships to R.J.L. (NS-8083) and to P.C. (Bourse Lavoisier and FRM, France).", "abstract": "Site-directed mutagenesis and expression in Xenopus oocytes were used to study acetylcholine receptors in which serine residues (i) were replaced by alanines (alpha, delta subunits) or (ii) replaced a phenylalanine (beta subunit) at a postulated polar site within the M2 transmembrane helix. As the number of serines decreased, there were decreases in the residence time and consequently the equilibrium binding affinity of QX-222, a quaternary ammonium anesthetic derivative thought to bind within the open channel. Receptors with three serine-to-alanine mutations also displayed a selective decrease in outward single-channel currents. Both the direction of this rectification and the voltage dependence of QX-222 blockade suggest that the residues mutated are within the aqueous pore of the receptor and near its cytoplasmic (inner) surface.", "date": "1988-12-16", "date_type": "published", "publication": "Science", "volume": "242", "number": "4885", "publisher": "American Association for the Advancement of Science", "pagerange": "1578-1581", "id_number": "CaltechAUTHORS:20150128-145454703", "issn": "0036-8075", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150128-145454703", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "NIH", "grant_number": "NS-8083" }, { "agency": "Bourse Lavoisier" }, { "agency": "Fondation pour la Recherche M\u00e9dicale" } ] }, "doi": "10.1126/science.2462281", "resource_type": "article", "pub_year": "1988", "author_list": "Leonard, Reid J.; Labarca, Cesar G.; et el." }, { "id": "https://authors.library.caltech.edu/records/ry3vr-ybn44", "eprint_id": 106035, "eprint_status": "archive", "datestamp": "2023-08-19 20:57:42", "lastmod": "2023-10-20 23:01:07", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Rudy-Bernardo", "name": { "family": "Rudy", "given": "Bernardo" } }, { "id": "Hoger-J-H", "name": { "family": "Hoger", "given": "Jeff H." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "At least two mRNA species contribute to the properties of rat brain A-type potassium channel expressed in xenopus oocytes", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1988 Cell Press. \n\nReceived 6 February 1988, Revised 30 July 1988. \n\nWe wish to thank Dr. R. Llinas for his support during this project; D. Krafte, T. Snutch, A. Coldin, J. Leonard, and R. J. Leonard for sharing their expertise; and J. Campanelli and A. Kamb for their interest and insightful comments and suggestions. This research was supported by USPHS grant GM26976 To B. R., GM10991 to N. D., and GM29836 to H. A. L. J. H. H. is a fellow of the American Cancer Society.", "abstract": "Fast transient K\u207a channels (A channels) of the type operating in the subthreshold region for Na\u207a action potential generation were expressed in Kenopus oocytes injected with rat brain poly(A) RNA. Sucrose gradient fractionation of the RNA separates mRNAs encoding A-currents (6\u20137 kb) from mRNAs encoding other voltage-dependent K\u207a channels. A-currents expressed with fractionated mRNA differ in kineticsAnd pharmacology from A-currents expressed with total mRNA. The original properties of the A-currents can be reconstituted when small mRNAs (2\u20134 kb) are added to the large mRNA fraction. Thus the properties of the A-currents expressed with total poly(A) RNA depend on the presence of more than one mRNA species. mRNA(s) present in the large RNA fraction must encode channel subunits since they express an A-current by themselves. The small mRNA(s) may encode a second subunit(s) or a factor, such as an enzymatic activity that modulates the properties of the channels, which could play a role in generating A-channel functional diversity.", "date": "1988-10", "date_type": "published", "publication": "Neuron", "volume": "1", "number": "8", "publisher": "Cell Press", "pagerange": "649-658", "id_number": "CaltechAUTHORS:20201013-140643992", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201013-140643992", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-26976" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "American Cancer Society" } ] }, "doi": "10.1016/0896-6273(88)90164-x", "resource_type": "article", "pub_year": "1988", "author_list": "Rudy, Bernardo; Hoger, Jeff H.; et el." }, { "id": "https://authors.library.caltech.edu/records/98sbk-v4e49", "eprint_id": 53115, "eprint_status": "archive", "datestamp": "2023-08-19 20:50:32", "lastmod": "2023-10-19 14:33:25", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Heterologous Expression of Excitability Proteins: Route to More Specific Drugs?", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1988 American Association for the Advancement of Science.\n\nI thank N. Davidson, T. M. Fong, H. Graves, D. S. Krafte, L. A. Jaffe, D. Julius,\nD. Kline, W. Koch, C. Labarca, R. J. Leonard, J. Lindstrom, E. McKenna, R. W.\nOlsen, J. H. Sabry, A. Schwartz, M. Tanouye, and L. Yu for comments on this\narticle and M. A. Buckles for assistance with the manuscript. Supported by grants\nfrom NIH, the Klingenstein Fund, and the Drown Foundation.", "abstract": "Many clinically important drugs act on the intrinsic membrane proteins (ion channels, receptors, and ion pumps) that control cell excitability. A major goal of pharmacology has been to develop drugs that are more specific for a particular subtype of excitability molecule. DNA cloning has revealed that many excitability proteins are encoded by multigene families and that the diversity of previously recognized pharmacological subtypes is matched, and probably surpassed, by the diversity of messenger RNAs that encode excitability molecules. In general, the diverse subtypes retain their properties when the excitability proteins are expressed in foreign cells such as oocytes and mammalian cell lines. Such heterologous expression may therefore become a tool for testing drugs against specific subtypes. In a systematic research program to exploit this possibility, major considerations include alternative processing of messenger RNA for excitability proteins, coupling to second-messenger systems, and expression of enough protein to provide material for structural studies.", "date": "1988-08-26", "date_type": "published", "publication": "Science", "volume": "241", "number": "4869", "publisher": "American Association for the Advancement of Science", "pagerange": "1057-1063", "id_number": "CaltechAUTHORS:20141222-150346710", "issn": "0036-8075", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20141222-150346710", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "Esther A. and Joseph Klingenstein Fund" }, { "agency": "Drown Foundation" } ] }, "doi": "10.1126/science.2457947", "resource_type": "article", "pub_year": "1988", "author_list": "Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/kd917-8zq70", "eprint_id": 5709, "eprint_status": "archive", "datestamp": "2023-08-22 05:56:07", "lastmod": "2023-10-23 17:19:28", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Iverson-L-E", "name": { "family": "Iverson", "given": "L. E." } }, { "id": "Tanouye-M-A", "name": { "family": "Tanouye", "given": "M. A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "N." } }, { "id": "Rudy-B", "name": { "family": "Rudy", "given": "B." } } ] }, "title": "A-Type Potassium Channels Expressed from Shaker Locus cDNA", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1988 by the National Academy of Sciences \n\nContributed by N. Davidson, April 1, 1988 \n\nWe thank D. Krafte for his expert advice in oocyte patch clamping, and J. Campanelli, A. Kamb, M. Mathew, J.D. Pollock, and M. Ramaswani for their comments. This research was supported by U.S. Public Health Service Grants GM26976 to B.R.; NS21327 to M.A.T., GM10991 to N.D., NS11756 to H.A.L., and a Pfeiffer Research Foundation grant to M.A.T. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - IVEpnas88b.pdf
", "abstract": "A-type K+ currents are expressed in Xenopus oocytes injected with in vitro-synthesized transcripts from cDNAs for the Drosophila Shaker (Sh) locus. A single Sh gene product, possibly as a multimer, is sufficient for formation of functional A channels. Various Sh RNAs express A currents with distinct kinetic properties. An analysis of structure-function relationships shows that the conserved central region of Sh polypeptides determines ionic selectivity and overall channel behavior, whereas the divergent amino and carboxyl termini can modify channel kinetics. Alternative splicing of Sh gene transcripts may provide one mechanism for the generation of K+ channel diversity.", "date": "1988-08-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "85", "number": "15", "publisher": "National Academy of Sciences", "pagerange": "5723-5727", "id_number": "CaltechAUTHORS:IVEpnas88b", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:IVEpnas88b", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM26976" }, { "agency": "NIH", "grant_number": "NS21327" }, { "agency": "NIH", "grant_number": "GM10991" }, { "agency": "NIH", "grant_number": "NS11756" }, { "agency": "Pfeiffer Research Foundation" } ] }, "pmcid": "PMC281833", "primary_object": { "basename": "IVEpnas88b.pdf", "url": "https://authors.library.caltech.edu/records/kd917-8zq70/files/IVEpnas88b.pdf" }, "resource_type": "article", "pub_year": "1988", "author_list": "Iverson, L. E.; Tanouye, M. A.; et el." }, { "id": "https://authors.library.caltech.edu/records/4xykn-3ce46", "eprint_id": 99645, "eprint_status": "archive", "datestamp": "2023-08-19 20:49:43", "lastmod": "2023-10-18 18:37:13", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Krafte-D-S", "name": { "family": "Krafte", "given": "Douglas S." } }, { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "Leonard-J-P", "name": { "family": "Leonard", "given": "John P." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Evidence for the involvement of more than one mRNA species in controlling the inactivation process of rat and rabbit brain Na channels expressed in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1988 Society for Neuroscience. \n\nReceived July 21, 1987; revised Jan. 15, 1988; accepted Jan. 18, 1988. \n\nWe wish to thank M. Fearey for technical assistance, F. Shon for assistance with the computer programs, and W. A. Catterall for the gift of a-scorpion toxin. \n\nThis work was supported by NS-11756, GM-10991, an NRSA individual fellowship (D.S.K.), and fellowships from the American Heart Association Greater Los Angeles Affiliate (T.P.S. and J.P.L.).\n\nPublished - 2859.full.pdf
", "abstract": "The properties of rat and rabbit brain sodium (Na) channels expressed in Xenopus oocytes following either unfractionated or high-molecular- weight mRNA injections were compared to assess the relative contribution of different size messages to channel function. RNA was size-fractionated on a sucrose gradient and a high-molecular-weight fraction (7\u201310 kilobase) encoding the \u03b1-subunit gave rise to functional voltage-dependent Na channels in the oocyte membrane. Single- channel conductance, mean open time, and time to first opening were all similar to the values for channels following injection of unfractionated RNA. In contrast, inactivation properties were markedly different; Na currents from high-molecular-weight RNA inactivated with a several-fold smaller macroscopic inactivation rate and showed a steady-state voltage dependence that was shifted in the depolarizing direction by at least 10 mV relative to that for unfractionated RNA. Single-channel recording revealed that the kinetic difference arose from a greater probability for high-molecular-weight RNA induced channels to reopen during a depolarizing voltage step. Pooling all gradient fractions and injecting this RNA into oocytes led to the appearance of Na channels with inactivation properties indistinguishable from those following injection of unfractionated RNA. These results suggest that mRNA species not present in the high- molecular-weight fraction can influence the inactivation process of rat brain Na channels expressed in Xenopus oocytes. This mRNA may encode \u03b2-subunits or other proteins that are involved in posttranslational processing of voltage-dependent Na channels.", "date": "1988-08-01", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "8", "number": "8", "publisher": "Society for Neuroscience", "pagerange": "2859-2868", "id_number": "CaltechAUTHORS:20191104-094627196", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191104-094627196", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "American Heart Association, Greater Los Angeles Affiliate" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1523/jneurosci.08-08-02859.1988", "pmcid": "PMC6569391", "primary_object": { "basename": "2859.full.pdf", "url": "https://authors.library.caltech.edu/records/4xykn-3ce46/files/2859.full.pdf" }, "resource_type": "article", "pub_year": "1988", "author_list": "Krafte, Douglas S.; Snutch, Terry P.; et el." }, { "id": "https://authors.library.caltech.edu/records/46n7k-44n52", "eprint_id": 106036, "eprint_status": "archive", "datestamp": "2023-08-19 20:48:53", "lastmod": "2023-10-20 23:01:13", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Auld-V-J", "name": { "family": "Auld", "given": "Vanessa J." } }, { "id": "Goldin-A-L", "name": { "family": "Goldin", "given": "Alan L." } }, { "id": "Krafte-D-S", "name": { "family": "Krafte", "given": "Douglas S." } }, { "id": "Marshall-John", "name": { "family": "Marshall", "given": "John" } }, { "id": "Dunn-J-M", "name": { "family": "Dunn", "given": "James M." } }, { "id": "Catterall-W-A", "name": { "family": "Catterall", "given": "William A." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Dunn-R-J", "name": { "family": "Dunn", "given": "Robert J." } } ] }, "title": "A rat brain Na\u207a channel \u03b1 subunit with novel gating properties", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1988 Cell Press. \n\nReceived 14 April 1988, Revised 25 May 1988. \n\nWe thank David Anderson for providing a rat brain cDNA library, Steve Clark for help with computer analysis, Shane Climie for sequencing advice, Jim Ingles for pEMBL-18, Derek van der Kooy for providing rat tissue, and Gayle Bodourian for manuscript preparation. This work was supported by the Medical Research Council of Canada (R. J. D.), National Institutes of Health Grants NS111756, and GM10991 (H. A. L. and N. D.), and the Muscular Dystrophy Association (H. A. L. and N. D.). A. L. G. is a Lucille P Markey Scholar, and this work was supported In part by a grant from the Lucille P. Markey Charitable Trust. D. S. K. is a Fellow of the National Institutes of Health.", "abstract": "We have constructed a full-length rat brain Na+ channel \u03b1 subunit cDNA that differs from the previously reported a subunit of Noda et al. at 6 amino acid positions. Transcription of the cDNA in vitro and injection into Xenopus oocytes resulted in the synthesis of functional Na\u207a channels. Although the single-channel conductance of the channels resulting from cloned cDNA was the same as that of channels resulting from injection of rat brain RNA, we observed two significant differences in the gating properties of the channels. The Na+ currents from cloned cDNA displayed much slower macroscopic inactivation compared with those from rat brain mRNA. In addition, the current-voltage relationship for currents from cloned cDNA was shifted 20\u201325 mV in the depolarizing direction compared with currents from rat brain RNA. Coinjection of low MW rat brain RNA restored normal inactivation of the channels indicating the presence of a component, either a structural subunit of the channel complex or a modifying enzyme, necessary for normal gating of the channel.", "date": "1988-08", "date_type": "published", "publication": "Neuron", "volume": "1", "number": "6", "publisher": "Cell Press", "pagerange": "449-461", "id_number": "CaltechAUTHORS:20201013-140644253", "issn": "0896-6273", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201013-140644253", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Medical Research Council of Canada" }, { "agency": "NIH", "grant_number": "NS-111756" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Lucille P. Markey Charitable Trust" }, { "agency": "NIH Postdoctoral Fellowship" } ] }, "doi": "10.1016/0896-6273(88)90176-6", "resource_type": "article", "pub_year": "1988", "author_list": "Auld, Vanessa J.; Goldin, Alan L.; et el." }, { "id": "https://authors.library.caltech.edu/records/mmdtz-q9031", "eprint_id": 106037, "eprint_status": "archive", "datestamp": "2023-08-22 05:47:51", "lastmod": "2023-10-20 23:01:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Sutton-Fedora", "name": { "family": "Sutton", "given": "Fedora" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Tetrodotoxin-sensitive voltage-dependent Na currents recorded from Xenopus oocytes injected with mammalian cardiac muscle RNA", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Xenopus oocyte; Tetrodotoxin; Cardiac; Ribonucleic acid", "note": "\u00a9 1988 Published by Elsevier. \n\nAccepted 22 September 1987. \n\nWe thank David J. Anderson for the gift of the neuronal specific clone SCG10; Kiyonori Yoshii for his assistance and advice with the initial electrophysiological experiments; Douglas Krafte for generous help and advice. This work was supported by NIH Grant HL 35782 and fellowships from the California Institute of Technology and the NIH to F.S.", "abstract": "Voltage-sensitive sodium (Na) channel currents recorded from mammalian cardiac muscle are blocked by tetrodotoxin (TTX) with a K_d of 1\u20133 \u03bcM. We have observed a K_d for TTX of 4\u201310 nM for Na currents recorded from Xenopus oocytes injected with RNA extracted from rabbit cardiac muscle. This result suggests that the degree of TTX sensitivity of Na channels encoded by cardiac muscle mRNA is in part determined by post-translational modification(s) or of associations with accessory proteins in the membrane.", "date": "1988-04", "date_type": "published", "publication": "Molecular Brain Research", "volume": "3", "number": "2", "publisher": "Elsevier", "pagerange": "187-191", "id_number": "CaltechAUTHORS:20201013-140644398", "issn": "0169-328X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201013-140644398", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "HL 35782" }, { "agency": "Caltech" }, { "agency": "NIH Postdoctoral Fellowship" } ] }, "doi": "10.1016/0169-328x(88)90065-4", "resource_type": "article", "pub_year": "1988", "author_list": "Sutton, Fedora; Davidson, Norman; et el." }, { "id": "https://authors.library.caltech.edu/records/5vyby-2he07", "eprint_id": 106152, "eprint_status": "archive", "datestamp": "2023-08-22 05:39:04", "lastmod": "2023-10-20 23:08:43", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Fong-Tung-Ming", "name": { "family": "Fong", "given": "Tung Ming" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Properties of two classes of rat brain acidic amino acid receptors induced by distinct mRNA populations in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "keywords": "RNA fractionation; Agonist interactions; Antagonist sensitivity", "note": "\u00a9 1988 Alan R. Liss, Inc. \n\nManuscript accepted: 25 July 1988; Manuscript received: 09 June 1988.", "abstract": "The Xenopus laevis oocyte expression system was used to study the molecular composition of mRNAs encoding acidic amino acid (AA) receptors from rat brain. Xenopus oocytes injected with poly(A) mRNA express two general classes of AA receptors. One class consists of AA\u2010gated cation channels. Responses are evoked by N\u2010methyl\u2010D\u2010aspartate (NMDA), by kainate, and to a lesser extent by L\u2010glutamate or quisqualate. The second class of receptor is coupled to an intracellular second messenger pathway activating an oocyte\u2010encoded Ca\u00b2\u207a\u2010activated Cl\u207b conductance. This second messenger\u2010coupled AA receptor can be activated by L\u2010glutamate or quisqualate. DL\u20102\u2010amino\u20105\u2010phosphonopentanoic acid and D\u2010\u03b1\u2010aminohexanedioic acid inhibit the AA\u2010gated cation conductances activated by NMDA or kainate with different potencies but do not inhibit the second messenger\u2010coupled AA receptor. Responses to NMDA are enhanced by micromolar level of glycine and are inhibited by Mg\u00b2\u207a, Zn\u00b2\u207a, or MK\u2010801. Dose\u2010response analysis reveals that the AA\u2010gated cation conductance activated by kainate requires the binding of two agonist molecules. To study the molecular composition, the mRNAs were size fractionated by denaturing agarose gel electrophoresis. About 20\u2010fold purification in specific activity (nA/ng of mRNA injected) of mRNAs encoding the second messenger coupled AA receptor was achieved. In contrast, only a slight enrichment of the mRNAs encoding the AA\u2010gated channel was observed. This suggests that the second messenger coupled AA receptor is encoded by a single size class of mRNA, whereas the AA\u2010gated cation channel(s) is encoded by multiple species of mRNAs or by mRNAs whose size distribution is heterogeneous.", "date": "1988", "date_type": "published", "publication": "Synapse", "volume": "2", "number": "6", "publisher": "Wiley", "pagerange": "657-665", "id_number": "CaltechAUTHORS:20201020-070235564", "issn": "0887-4476", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201020-070235564", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1002/syn.890020613", "resource_type": "article", "pub_year": "1988", "author_list": "Fong, Tung Ming; Davidson, Norman; et el." }, { "id": "https://authors.library.caltech.edu/records/t1fdc-mzb59", "eprint_id": 32047, "eprint_status": "archive", "datestamp": "2023-08-19 19:51:25", "lastmod": "2023-10-17 22:23:26", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Yoshii-Kiyonori", "name": { "family": "Yoshii", "given": "Kiyonori" } }, { "id": "Yu-Lei", "name": { "family": "Yu", "given": "Lei" } }, { "id": "Mayne-K-M", "name": { "family": "Mayne", "given": "Katharine Mixter" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Equilibrium Properties of Mouse-Torpedo Acetylcholine Receptor Hybrids Expressed in Xenopus Oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1987 Rockefeller University Press. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. \n\nOriginal version received 10 March 1987; accepted version received 16 June 1987; Published October 1, 1987. \n\nWe thank Dr. J. P. Merlie for providing clones and Dr. Reid Leonard for discussion.\nThis research was supported by grants from the Muscular Dystrophy Association and from the National Institutes of Health (NS-11752).\n\nPublished - YOSjgp87.pdf
", "abstract": "This study used messenger RNA encoding each subunit (\u03b1, \u03b2, \u03b3\nand \u03b4) of the nicotinic acetylcholine (ACh) receptor from mouse BC3H-1 cells and from Torpedo electric organ. The mRNA was synthesized in vitro by transcription with SP6 polymerase from cDNA clones. All 16 possible combinations that include one mRNA for each of \u03b1, \u03b2, \u03b3 and \u03b4 were injected into oocytes. After allowing 2-8 d for translation and assembly, we assayed each oocyte for (a) receptor assembly, measured by the binding of [^125]\u03b1-bungarotoxin to the oocyte surface, and (b) ACh-induced conductance, measured under voltage clamp at various membrane potentials. All combinations yielded detectable\nassembly (30-fold range among different combinations) and ACh-induced\nconductances (>1,000-fold range at 1 \u00b5M). On double-logarithmic coordinates, the dose-response relations all had a slope near 2 for low concentrations of ACh. Data were corrected for variations in efficiency of translation among identically injected oocytes by expressing ACh-induced conductance per femtomole\nof \u03b1-bungarotoxin-binding sites. Five combinations were tested for d-tubocurarine\ninhibition by the dose-ratio method; the apparent dissociation\nconstant ranged from 0.08 to 0.27 \u00b5M. Matched responses and geometric\nmeans are used for describing the effects of changing a particular subunit\n(mouse vs. Torpedo) while maintaining the identity of the other subunits. A\ndramatic subunit-specific effect is that of the \u03b2 subunit on voltage sensitivity of\nthe response: g_ACh(-90 mV)/g_Ach(+30 mV) is always at least 1, but this ratio\nincreases by an average of 3.5-fold if \u03b2_M replaces \u03b2_T. Also, combinations\nincluding \u03b3_T or \u03b4_M usually produce greater receptor assembly than combinations\nincluding the homologous subunit from the other species. Finally, E_ACh is\ndefined as the concentration of ACh inducing 1 \u00b5S/fmol at -60 mV; E_ACh is\nconsistently lower for \u03b1_m. We conclude that receptor assembly, voltage sensitivity,\nand E_ACh are governed by different properties.", "date": "1987-10-01", "date_type": "published", "publication": "Journal of General Physiology", "volume": "90", "number": "4", "publisher": "Rockefeller University Press", "pagerange": "553-573", "id_number": "CaltechAUTHORS:20120622-143923786", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120622-143923786", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Muscular Dystrophy Association" }, { "agency": "NIH", "grant_number": "NS-11752" } ] }, "doi": "10.1085/jgp.90.4.553", "pmcid": "PMC2228871", "primary_object": { "basename": "YOSjgp87.pdf", "url": "https://authors.library.caltech.edu/records/t1fdc-mzb59/files/YOSjgp87.pdf" }, "resource_type": "article", "pub_year": "1987", "author_list": "Yoshii, Kiyonori; Yu, Lei; et el." }, { "id": "https://authors.library.caltech.edu/records/dcjmm-nz288", "eprint_id": 106038, "eprint_status": "archive", "datestamp": "2023-08-22 05:27:03", "lastmod": "2023-10-20 23:01:25", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Mayne-K-M", "name": { "family": "Mayne", "given": "Katharine Mixter" } }, { "id": "Yoshii-Kiyonori", "name": { "family": "Yoshii", "given": "Kiyonori" } }, { "id": "Yu-Lei", "name": { "family": "Yu", "given": "Lei" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Expression of mouse-Torpedo acetylcholine receptor subunit chimeras and hybrids in Xenopus oocytes", "ispublished": "pub", "full_text_status": "restricted", "keywords": "In vitro transcription; Ion channel", "note": "\u00a9 1987 Published by Elsevier. \n\nAccepted 5 May 1987. \n\nWe thank Drs. T. Claudio, S. Heinemann, and D. Noonan for kindly providing Torpedo AChR cDNA clones, Drs. D. Mead and D. Melton for providing SP6 expression plasmids and information concerning the SP6 transcription system, Drs. J. Jackson and E. Barnard for the chick a cDNA clone, Dr. J.P. Merlie for the mouse a cDNA clone, and Drs. R. Stroud and J. Finer-Moore for helpful discussions. This work has been supported in part by research grants GM-10991 and NS-11756 from the National Institutes of Health, a research grant for the Muscular Dystrophy Association, support from the government of Japan for K.Y., Predoctoral Training Grant GM-07616 from the National Institutes of Health to K.M.M., and a California Foundation for Biochemical Research Fellowship to L.Y.", "abstract": "In this study, in vitro synthesized mRNA encoding mouse and Torpedo nicotinic acetylcholine receptor subunits was injected into Xenopus oocytes, followed by assays for assembly onto the oocyte surface (using [\u00b9\u00b2\u2075I]\u03b1-bungarotoxin binding) and for acetylcholine-induced conductances (using voltage clamp). We constructed hybrid acetylcholine receptors in Xenopus oocytes by injecting all 8 possible combinations of 4 subunit-specific mRNAs in which a single subunit is derived from the other species. For each hybrid combination, there is detectable assembly and conductance. We also constructed cDNA clones that encode chimeric acetylcholine receptor subunits in which part of the \u03b3 subunit from Torpedo was replaced by the homologous region of the \u03b4 subunit from mouse. None of the chimeric subunits was able to replace the Torpedo \u03b3, mouse \u03b4, or Torpedo \u03b4 subunit with regard to assembly or function. We therefore conclude that widely spaced (and unknown) parts of the protein chain are required for the intersubunit interactions that eventually lead to functional assembly of the receptor.", "date": "1987-09", "date_type": "published", "publication": "Molecular Brain Research", "volume": "2", "number": "3", "publisher": "Elsevier", "pagerange": "191-197", "id_number": "CaltechAUTHORS:20201013-140644515", "issn": "0169-328X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201013-140644515", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Ministry of Education, Culture, Sports, Science and Technology (MEXT)" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM-07616" }, { "agency": "California Foundation for Biochemical Research" } ] }, "doi": "10.1016/0169-328x(87)90026-x", "resource_type": "article", "pub_year": "1987", "author_list": "Mayne, Katharine Mixter; Yoshii, Kiyonori; et el." }, { "id": "https://authors.library.caltech.edu/records/gf8ej-98x21", "eprint_id": 6751, "eprint_status": "archive", "datestamp": "2023-08-22 05:21:43", "lastmod": "2023-10-23 16:15:32", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "L\u00fcbbert-H", "name": { "family": "L\u00fcbbert", "given": "Hermann" } }, { "id": "Hoffman-B-J", "name": { "family": "Hoffman", "given": "Beth J." } }, { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "van-Dyke-T", "name": { "family": "van Dyke", "given": "Terry" } }, { "id": "Levine-A-J", "name": { "family": "Levine", "given": "Arnold J." } }, { "id": "Hartig-P-R", "name": { "family": "Hartig", "given": "Paul R." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "cDNA Cloning of a Serotonin 5-HT1C Receptor by Electrophysiological Assays of mRNA-Injected Xenopus Oocytes", "ispublished": "pub", "full_text_status": "public", "keywords": "RNA fractionation; hybrid depletion; hybrid selection; choroid plexus; voltage clamp", "note": "\u00a9 1987 by the National Academy of Sciences \n\nContributed by Norman Davidson, March 16, 1987 \n\nWe thank Hieu Nguyen for technical assistance, J. Vieira and J. Messing for supplying pUC119 and M13K07, and J. Kowalski and D.T. Denhardt for donating the bacterial strain E. coli R4. This research has been supported by National Institutes of Health grants GM-10991, NS-11756, NS-23048, and CA-38757 and by fellowship support from the Deutsche Forschungsgemeinschaft and the American Cancer Society, California Division to H.L.; by the National Science Foundation to B.J.H.; by the American Heart Association, Greater Los Angeles Affiliate, and the Natural Sciences and Engineering Research Council of Canada to T.P.S. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - LUBpnas87.pdf
", "abstract": "We describe a strategy for the cloning of neurotransmitter-receptor and ion-channel cDNAs that is based on electrophysiological assays of mRNA-injected Xenopus oocytes. This procedure circumvents the purification of these membrane proteins, which is hindered by their low abundance and their hydrophobic nature. It involves methods for RNA fractionation by high-resolution gel electrophoresis, directional cDNA cloning in a single-stranded vector, and screening of the cDNA library by voltage-clamp measurements of currents induced by serotonin in mRNA-injected oocytes. The applicability of our approach is demonstrated by the isolation of a serotonin receptor cDNA clone from a mouse choroid plexus papilloma. The clone was identified by hybrid-depletion and hybrid-selection procedures. The receptor expressed in oocytes injected with hybrid-selected RNA is fully functional, indicating that it is composed of a single subunit encoded by a 5-kilobase RNA. The pharmacology of the hybrid-selected receptor confirms that we have successfully cloned a serotonin 5-HT1C receptor cDNA.", "date": "1987-06-15", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "84", "number": "12", "publisher": "National Academy of Sciences", "pagerange": "4332-4336", "id_number": "CaltechAUTHORS:LUBpnas87", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LUBpnas87", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-23048" }, { "agency": "NIH", "grant_number": "CA-38757" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)" }, { "agency": "American Cancer Society, California Division" }, { "agency": "American Heart Association, Greater Los Angeles Affiliate" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" } ] }, "pmcid": "PMC305079", "primary_object": { "basename": "LUBpnas87.pdf", "url": "https://authors.library.caltech.edu/records/gf8ej-98x21/files/LUBpnas87.pdf" }, "resource_type": "article", "pub_year": "1987", "author_list": "L\u00fcbbert, Hermann; Hoffman, Beth J.; et el." }, { "id": "https://authors.library.caltech.edu/records/kbg92-11875", "eprint_id": 5805, "eprint_status": "archive", "datestamp": "2023-08-22 05:20:13", "lastmod": "2023-10-23 16:15:25", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Gurney-A-M", "name": { "family": "Gurney", "given": "Alison M." } }, { "id": "Tsien-R-Y", "name": { "family": "Tsien", "given": "Roger Y." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Activation of a Potassium Current by Rapid Photochemically Generated Step Increases of Intracellular Calcium in Rat Sympathetic Neurons", "ispublished": "pub", "full_text_status": "public", "keywords": "light flash; calcium jump; superior cervical ganglion", "note": "\u00a9 1987 by the National Academy of Sciences \n\nCommunicated by Melvin I. Simon, January 20, 1987 \n\nWe thank Dr. S. Adams for synthesizing nitr-5 and nitr-7, Dr. J.M. Nerbonne for advice and helpful discussions, for participating in early experiments, and for providing facilities for some of the later experiments. We also thank Dr. C. Miller for a gift of charybdotoxin and for much discussion, Drs. P.R. Adams, R. Brett, S. Jones, J. Nargeot, and R. Zucker for discussion; and B. Tanamachi, D. Martin, and J. Doyle for preparing the cells. This research was supported by the American Heart Association Postdoctoral Fellowship to A.M.G., by Grant 83-K-111 from the Searle Scholars Program, by Grants GM-29836, GM-31004, and EY04372 from the National Institutes of Health, and by the Medical Research Council and British Heart Foundation. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - GURpnas87.pdf
", "abstract": "Although Ca2+ is a well-established intracellular messenger, there are many questions concerning the kinetics and spatial localization of its effects. Such problems may now be approached with the photosensitive Ca2+ chelator nitr-5. The Ca2+ affinity of this molecule decreases by a factor of 40 after absorption of near-UV light; Ca2+ is liberated with a time constant of approx 300 \u00b5s. Nitr-5 or the related compounds nitr-2 and nitr-7, complexed with Ca2+, were introduced into rat sympathetic ganglion cells by dialysis from a patch pipette electrode operating in the whole-cell, voltage-clamp mode. Light flashes released Ca2+ and activated a K+ current. Flash-induced current relaxations followed a simple exponential time course with time constants as brief as 5 ms. Comparison of the kinetics among the chelators, which photolyze at different rates, suggests that release of Ca2+ from nitr-5 is too fast to limit the relaxation. Thus we confirm directly that Ca2+ can modulate membrane properties within a few milliseconds after entering a cell. A preliminary kinetic description of K+ current activation by Ca2+ in rat sympathetic neurons is presented; Ca2+ appears to bind to the channel with a rate constant of at least 2 x 10^7 M^-1\u00b7 s^-1.", "date": "1987-05-15", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "84", "number": "10", "publisher": "National Academy of Sciences", "pagerange": "3496-3500", "id_number": "CaltechAUTHORS:GURpnas87", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:GURpnas87", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "American Heart Association" }, { "agency": "Searle Scholars Program", "grant_number": "83-K-111" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "GM-31004" }, { "agency": "NIH", "grant_number": "EY04372" }, { "agency": "Medical Research Council (UK)" }, { "agency": "British Heart Foundation" } ] }, "pmcid": "PMC304898", "primary_object": { "basename": "GURpnas87.pdf", "url": "https://authors.library.caltech.edu/records/kbg92-11875/files/GURpnas87.pdf" }, "resource_type": "article", "pub_year": "1987", "author_list": "Gurney, Alison M.; Tsien, Roger Y.; et el." }, { "id": "https://authors.library.caltech.edu/records/fe17a-2d189", "eprint_id": 106039, "eprint_status": "archive", "datestamp": "2023-08-19 19:29:36", "lastmod": "2023-10-20 23:01:28", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Gurney-A-M", "name": { "family": "Gurney", "given": "Alison M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Light-flash physiology with synthetic photosensitive compounds", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1987 American Physiological Society. \n\nWe thank A. Cuthbert, J. M. Nerbonne, D. Trentham, R. Y. Tsien, and our many other colleagues and collaborators over the past 10 years for advice, encouragement, and ideas. \n\nThe research at California Institute of Technology has been supported by grants from the National Institutes of Health (NS-11756 and GM-29836) and by fellowships and grants from the American Heart Association and the Muscular Dystrophy Association. \n\nThe literature search for this review was completed in July 1986.\n\nPublished - physrev.1987.67.2.583.pdf
", "abstract": "This review treats a technique that has proved useful recently in some physiological studies. A process is measured while a light flash is used to manipulate a physiologically important or pharmacologically active photosensitive molecule. Typically such experiments complement more common measurements of 1) equilibrium responses in the same system or 2) relaxations after voltage jumps, rapid mixing, and so on. The chief advantage of the lightflash technique is the speed of the photochemistry and the fact that it can be applied to organized systems, such as muscle fibers and membranes under electrophysiological investigation, that cannot be flowed. Experiments may be classified by the accessible time scales (Table 1), by successful recording techniques (Table 2), and by available molecules (Table 3).", "date": "1987-04", "date_type": "published", "publication": "Physiological Reviews", "volume": "67", "number": "2", "publisher": "American Physiological Society", "pagerange": "583-617", "id_number": "CaltechAUTHORS:20201013-140644623", "issn": "0031-9333", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201013-140644623", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "American Heart Association" }, { "agency": "Muscular Dystrophy Association" } ] }, "doi": "10.1152/physrev.1987.67.2.583", "primary_object": { "basename": "physrev.1987.67.2.583.pdf", "url": "https://authors.library.caltech.edu/records/fe17a-2d189/files/physrev.1987.67.2.583.pdf" }, "resource_type": "article", "pub_year": "1987", "author_list": "Gurney, Alison M. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/6ak2s-7ed60", "eprint_id": 106040, "eprint_status": "archive", "datestamp": "2023-08-19 19:29:41", "lastmod": "2023-10-20 23:01:31", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "L\u00fcbbert-H", "name": { "family": "L\u00fcbbert", "given": "Hermann" } }, { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Rat brain 5-HT_(1C) receptors are encoded by a 5-6 kbase mRNA size class and are functionally expressed in injected Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1987 by Society for Neuroscience. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). \n\nReceived June 30, 1986; revised Oct. 16, 1986; accepted Oct. 17, 1986. \n\nThe authors wish to thank Stephen J. Peroutka, Terry T. Takahashi, and Mary B. Kennedy for advice in receptor pharmacology and brain anatomy. We are grateful to Stephen J. Peroutka for donating RU 24969 and mesulergine, to Paul R. Hartig for supplying ketanserin, and to Beth J. Hoffman for critically reading the manuscript. This work was supported by NIH Grants GM-10991 and NS-11756 and by fellowships from the Deutsche Forschungsgemeinschaft to H.L., from the American Heart Association, Greater Los Angeles Affiliate, and NSERC of Canada to T.P.S., and from the Bantrell and Fulbright Foundations to N.D.\n\nPublished - 1159.full.pdf
", "abstract": "Injection of rat brain RNA into Xenopus laevis oocytes induces synthesis of receptors that show an electrophysiological response to bath application of serotonin. While there are at least 4 pharmacologically distinct subtypes of 5-HT binding sites in the rat brain, we find that the pharmacological characteristics of the predominant electrophysiologically active receptor synthesized in Xenopus oocytes are most consistent with those of the 5-HT_(1C) subtype. Additional electrophysiologically active 5-HT receptor types could not be detected. Injection of mRNA isolated from a number of rat brain regions shows that the choroid plexus is particularly enriched for 5-HT_(1C) mRNA. Oocytes injected with RNA isolated from this region respond 16 or 8 times more strongly to serotonin than do oocytes injected with RNA isolated from cortex or substantia nigra, respectively. In addition, by fractionation of rat brain mRNA through agarose gels, we have identified a single RNA size class of about 5\u20136 kbase that encodes this serotonin receptor.", "date": "1987-04", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "7", "number": "4", "publisher": "Society for Neuroscience", "pagerange": "1159-1165", "id_number": "CaltechAUTHORS:20201013-145241211", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201013-145241211", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)" }, { "agency": "American Heart Association, Greater Los Angeles Affiliate" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "Fulbright Foundation" } ] }, "doi": "10.1523/jneurosci.07-04-01159.1987", "pmcid": "PMC6569006", "primary_object": { "basename": "1159.full.pdf", "url": "https://authors.library.caltech.edu/records/6ak2s-7ed60/files/1159.full.pdf" }, "resource_type": "article", "pub_year": "1987", "author_list": "L\u00fcbbert, Hermann; Snutch, Terry P.; et el." }, { "id": "https://authors.library.caltech.edu/records/jsw59-djg40", "eprint_id": 106041, "eprint_status": "archive", "datestamp": "2023-08-19 19:25:48", "lastmod": "2023-10-20 23:01:34", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Leonard-J-P", "name": { "family": "Leonard", "given": "John P." } }, { "id": "Nargeot-J", "name": { "family": "Nargeot", "given": "Jo\u00ebl" } }, { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Ca channels induced in Xenopus oocytes by rat brain mRNA", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1987 by Society for Neuroscience. Beginning six months after publication the Work will be made freely available to the public on SfN's website to copy, distribute, or display under a Creative Commons Attribution 4.0 International (CC BY 4.0) license (https://creativecommons.org/licenses/by/4.0/). \n\nReceived June 20, 1986; accepted Sept. 13, 1986. \n\nWe thank N. Dascal, D. S. Krafte, and R. J. Miller for helpful suggestions. This research was supported by fellowships from the American Heart Association, Greater Los Angeles Affiliate (to J.L. and T.S.), the Conseil de recherches en sciences naturelles et en genie du Canada (to T.S.), and the National Institutes of Health (Grants GM-10991 and GM-29836).\n\nPublished - 875.full.pdf
", "abstract": "RNA was isolated from brains of 16-d-old rats and poly(A) samples were injected into stage V and VI oocytes. After allowing 2\u20135 d for expression, most oocytes were exposed to medium in which the K had been replaced by Cs for 24 hr prior to recording. Ba currents were usually measured in Cl-free Ba-methanesulfonate saline. \n\nI_(Ba) in noninjected oocytes was often undetectable, but ranged up to 50 nA (22 \u00b1 4 nA, n = 21). In contrast, injected oocytes showed a peak I_(Ba) of 339 \u00b1 42 nA (n = 33). The threshold for activation of I_(Ba) was -40 mV, with peak currents at +10 to +20 mV. After a peak, currents decayed to a nearly steady level along a single-exponential time course (\u03c4 = 650 \u00b1 50 msec at +20 mV). The maintained current was 67 \u00b1 6% (n = 9) of the early peak amplitude. A prepulse duration of 5 sec was needed to examine the inactivation of barium currents in injected oocytes. The inward I_(Ba) could be observed in BaCl\u2082 solutions at potentials positive to E_(Cl) and also in Na-free salines, indicating that neither Cl\u207b nor Na\u207a was carrying the inward current. \n\nAlthough I_(Ba) displayed voltage- independent blockade by Cd (50% inhibition at 6 \u00b5M), the peptide Ca channel antagonist, \u03c9-CgTX (1 \u00b5M), and the organic Ca channel-blocking agents (verapamil, compound W-7, and nifedipine) were uniformly ineffective. No effects were observed with the dihydropyridine antagonist nifedipine (even at 10 \u00b5M, or when cells were held at -40 mV) or agonist Bay K-8644. However, I_(Ba) was enhanced via activation of protein kinase C with 4-beta-phorbol dibutyrate (PBT\u2082). In contrast, use of forskolin to activate protein kinase A did not alter I_(Ba). However, experiments in the presence of Cd revealed that forskolin decreased I_K. Ca channels produced by rat brain mRNA were thus in contrast to the nifedipine-sensitive, Bay K-8644- and forskolin-enhanced Ca channels observed after injection of rat heart mRNA (Dascal et al., 1986).", "date": "1987-03", "date_type": "published", "publication": "Journal of Neuroscience", "volume": "7", "number": "3", "publisher": "Society for Neuroscience", "pagerange": "875-881", "id_number": "CaltechAUTHORS:20201013-145241336", "issn": "0270-6474", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201013-145241336", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "American Heart Association, Greater Los Angeles Affiliate" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "GM-29836" } ] }, "doi": "10.1523/jneurosci.07-03-00875.1987", "pmcid": "PMC6569056", "primary_object": { "basename": "875.full.pdf", "url": "https://authors.library.caltech.edu/records/jsw59-djg40/files/875.full.pdf" }, "resource_type": "article", "pub_year": "1987", "author_list": "Leonard, John P.; Nargeot, Jo\u00ebl; et el." }, { "id": "https://authors.library.caltech.edu/records/63efz-h8f54", "eprint_id": 105976, "eprint_status": "archive", "datestamp": "2023-08-19 19:13:31", "lastmod": "2023-10-20 22:56:16", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "Leonard-J-P", "name": { "family": "Leonard", "given": "John P." } }, { "id": "Nargeot-J", "name": { "family": "Nargeot", "given": "Jo\u00ebl" } }, { "id": "L\u00fcbbert-H", "name": { "family": "L\u00fcbbert", "given": "Hermann" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Characterization of voltage-gated calcium channels in Xenopus oocytes after injection of RNA from electrically excitable tissues", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1987 Raven Press.\n\nWe thank Dr. Douglas Krafte and Dr. Nathan Dascal for their comments on this manuscript. We also thank Moira Fearey for excellent technical assistance.\nThis research was supported by fellowships from the American Heart Association Greater Los Angeles Affiliate, and the Natural Science and Engineering Research Council of Canada, and by grants GM-10991, GM-29836, and HL-35782 from the National Institutes of Health. J.N. thanks Laboratoire Servier (Neuilly-sur-Seine) for a travel grant.", "abstract": "The entry of Ca\u00b2\u207a through voltage-gated channels has two major functions. First, Ca\u00b2\u207a fluxes elevate the intracellular concentration of this important second messenger. Second, Ca\u00b2\u207a currents directly influence membrane potential, thereby contributing to impulse patterns. Ca\u00b2\u207a channels serve these functions in a variety of nerve, muscle, and endocrine cells (Reuter, 1983; Tsien, 1983). As might be expected from their wide distribution and their role in regulating a number of cellular functions, voltage-gated Ca\u00b2\u207a channels form a heterogeneous family. Diverse types of Ca\u00b2\u207a channels can be distinguished with regard to gating kinetics, pharmacology, and permeability (Fox and Krasne, 1981; Armstrong and Matteson, 1985; Nowyeky et al., 1985). For the purpose of characterizing the various forms of voltage-gated Ca\u00b2\u207a channels, it would be desirable to study them in a similar membrane environment.", "date": "1987", "date_type": "published", "publication": "Society of General Physiologists series", "volume": "42", "publisher": "Raven Press", "pagerange": "153-166", "id_number": "CaltechAUTHORS:20201009-153923021", "issn": "0094-7733", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201009-153923021", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "American Heart Association" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "HL-35782" }, { "agency": "Laboratoires Servier" } ] }, "resource_type": "article", "pub_year": "1987", "author_list": "Snutch, Terry P.; Leonard, John P.; et el." }, { "id": "https://authors.library.caltech.edu/records/ek9g9-dhg92", "eprint_id": 105992, "eprint_status": "archive", "datestamp": "2023-08-22 05:04:07", "lastmod": "2023-10-20 22:57:33", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Ifune-Catherine", "name": { "family": "Ifune", "given": "Catherine" } }, { "id": "Hopkins-R-S", "name": { "family": "Hopkins", "given": "Rosemary" } }, { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "L\u00fcbbert-H", "name": { "family": "L\u00fcbbert", "given": "Hermann" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Simon-M-I", "name": { "family": "Simon", "given": "Melvin I." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Involvement of a GTP-binding protein in mediation of serotonin and acetylcholine responses in Xenopus oocytes injected with rat brain messenger RNA", "ispublished": "pub", "full_text_status": "restricted", "keywords": "Inositol trisphosphate; Chloride channel; Calcium", "note": "\u00a9 1986 Published by Elsevier. \n\nAccepted 9 September 1986. \n\nThis work was supported by NIH Grants GM-29836 to H.A.L., GM-10991 to N.D., and GM-34236 to M.S. N.D. was a recipient of Bantrell and Fulbright Postdoctoral Fellowships. H.L. was a fellow of the Deutsche Forschungsgemeinschaft. T.S. was a postdoctoral fellow of the American Heart Association/Greater Los Angeles Affiliate. The authors are grateful to Ms. Moira Fearey for excellent technical assistance.", "abstract": "Injection of poly(A)\u207a RNA from rat brain into Xenopus oocytes caused the appearance of Cl currents in response to serotonin (5-HT) and acetylcholine (ACh). Both neurotransmitters evoked two-component currents similar in their time course to the oocyte's endogenous cholinergic muscarinic response, which was shown in previous studies to be mediated by IP\u2083 synthesis leading to Ca release from intracellular stores. The responses to ACh and 5-HT exhibited self- and cross-desensitization, i.e., application of either ACh or 5-HT inhibited the subsequent response to either one of the two transmitters. Intracellular injection of guanosine 5\u2032-O-(3-thiotriphosphate) (GTP-\u03b3-S) mimicked the 5-HT and ACh response, and also completely suppressed the response to the subsequent application of either ACh or 5-HT. Treatment of the oocytes with pertussis toxin (PTX) caused a 50% attenuation of ACh and 5-HT responses. In the membranes of both control and mRNA-injected oocytes, PTX catalyzed the ADP-ribosylation of a single M_r = \u223c40,000 protein. Injection of the purified \u03b2\u03b3-subunits of transducin enhanced the 5-HT response. The 5-HT and GTP-\u03b3-S responses were inhibited by intracellular injection of the Ca\u00b2\u207a chelator, EGTA, as previously shown for the ACh response. These data suggest that ACh and 5-HT receptors, synthesized in the oocytes on the template of brain mRNA, act through a common pathway that involves (a) a guanine nucleotide binding protein and (b) IP\u2083 production leading to Ca mobilization.", "date": "1986-12", "date_type": "published", "publication": "Molecular Brain Research", "volume": "1", "number": "3", "publisher": "Elsevier", "pagerange": "201-209", "id_number": "CaltechAUTHORS:20201012-152225953", "issn": "0169-328X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-152225953", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "GM-34236" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "Fulbright Foundation" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)" }, { "agency": "American Heart Association, Greater Los Angeles Affiliate" } ] }, "doi": "10.1016/0169-328x(86)90026-4", "resource_type": "article", "pub_year": "1986", "author_list": "Dascal, Nathan; Ifune, Catherine; et el." }, { "id": "https://authors.library.caltech.edu/records/qmdt2-jns87", "eprint_id": 105993, "eprint_status": "archive", "datestamp": "2023-08-19 18:53:19", "lastmod": "2023-10-20 22:57:40", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Chabala-L-D", "name": { "family": "Chabala", "given": "Lee D." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Activation of acetylcholine receptor channels by covalently bound agonists in cultured rat myoballs", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1986 The Physiological Society. \n\n(Received 21 May 1985) \n\nWe thank C. Ifune for participating in some of the experiments, Dr M. A. Raftery for supplying BrACh, and Drs N. H. Wassermann and B. F. Erlanger for supplying QBr. This research was supported by a fellowship from the Muscular Dystrophy Association of America to L. D. C. and by a grant from the NIH (NS-11756).", "abstract": "Kinetic and equilibrium aspects of receptor activation by two irreversibly bound ('tethered') agonists, QBr and bromoacetylcholine (BrACh), were examined in cultured embryonic rat muscle. Myoballs were treated with dithiothretitol (2 mM), washed, exposed to BrACh or QBr, and then washed again. Voltage\u2010clamp recordings were made both in the whole\u2010cell mode and with excised outside\u2010out patches at 15 degrees C. Whole\u2010cell voltage\u2010jump relaxations resembled those observed with reversibly bound agonists. The relaxation time constants were 5 ms for tethered QBr and 10 ms for tethered BrACh (\u2010100 mV, 15 degrees C). At more positive membrane potentials, the relaxation rate constants increased and the conductance decreased. Whole\u2010cell light\u2010flash relaxations with tethered QBr were also studied. The conductance was increased and decreased, respectively, by cis\u2192trans and trans\u2192cis photoisomerizations. The relaxation time constants equalled those for voltage jumps. The functional stoicheiometry of tethered QBr was investigated by studying the relaxations in response to light flashes that produced known changes in the mole fractions of the two isomers. It is concluded that the open state of each receptor channel is controlled by the isomeric state of a single tethered QBr molecule. In single\u2010channel recordings, tethered agonists opened channels with the same conductance as reversibly bound agonists (30 pS at 15 degrees C and \u2010100 mV). More than 80% of the conductance was contributed by a population of openings with an average burst duration (lifetime) of 5 ms for QBr and 10 ms for BrACh. Thus the single\u2010channel and macroscopic currents seem to be dominated by the same type of channel; these are presumably monoliganded receptors. About 30% of the openings belonged to a population with an average lifetime of about 0.5 ms. This population contributed less than 5% of the conductance. There were also more long openings (greater than 50 ms) than expected from a simple exponential distribution. A few patches from BrACh\u2010treated cells showed openings with a conductance of 45 pS (\u2010100 mV) and an average duration of approximately 2 ms. These data allow one to assess whether the agonist\u2010receptor binding step plays a role in generating the brief openings. The main population of openings (burst durations 5 ms with QBr and 10 ms with BrACh) seem to be contributed by monoliganded receptors. One can therefore rule out the hypothesis that the brief channels arise exclusively from mono\u2010 and biliganded receptors, respectively.", "date": "1986-10-01", "date_type": "published", "publication": "Journal of Physiology", "volume": "379", "number": "1", "publisher": "Physiological Society", "pagerange": "83-108", "id_number": "CaltechAUTHORS:20201012-152226105", "issn": "0022-3751", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-152226105", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Muscular Dystrophy Association of America" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1113/jphysiol.1986.sp016242", "pmcid": "PMC1182886", "resource_type": "article", "pub_year": "1986", "author_list": "Chabala, Lee D. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/e1krd-76h54", "eprint_id": 11845, "eprint_status": "archive", "datestamp": "2023-08-22 05:00:28", "lastmod": "2023-10-17 15:49:50", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Goldin-A-L", "name": { "family": "Goldin", "given": "Alan L." } }, { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "L\u00fcbbert-H", "name": { "family": "L\u00fcbbert", "given": "Hermann" } }, { "id": "Dowsett-A", "name": { "family": "Dowsett", "given": "Andrew" } }, { "id": "Marshall-J", "name": { "family": "Marshall", "given": "John" } }, { "id": "Auld-V-J", "name": { "family": "Auld", "given": "Vanessa" } }, { "id": "Downey-W", "name": { "family": "Downey", "given": "William" } }, { "id": "Fritz-L-C", "name": { "family": "Fritz", "given": "Larry C." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Dunn-R-J", "name": { "family": "Dunn", "given": "Robert" } }, { "id": "Catterall-W-A", "name": { "family": "Catterall", "given": "William A." } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Messenger RNA coding for only the alpha subunit of the rat brain Na channel is sufficient for expression of functional channels in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "keywords": "hybrid selection; sucrose-gradient fractionation; voltage clamp", "note": "\u00a9 1986 by the National Academy of Sciences. \n\nContributed by Norman Davidson, October 28, 1985. \n\nWe thank Nathan Dascal for his expert assistance and advice in performing the electrophysiological procedures. This research has been supported by grants from the National Institutes of Health to N.D., W.A.C., and H.A.L.; by a grant from the Medical Research Council of Canada to R.D.; and by fellowship support from the American Heart Association to A.D. and T.S.; from the Natural Sciences and Engineering Research Council of Canada to T.S.; from the Deutsche Forschungsgemeinschaft to H.L.; and from the National Multiple Sclerosis Society to A.L.G. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - GOLpnas86.pdf
", "abstract": "Several cDNA clones coding for the high molecular weight (alpha) subunit of the voltage-sensitive Na channel have been selected by immunoscreening a rat brain cDNA library constructed in the expression vector lambda gt11. As will be reported elsewhere, the amino acid sequence translated from the DNA sequence shows considerable homology to that reported for the Electrophorus electricus electroplax Na channel. Several of the cDNA inserts hybridized with a low-abundance 9-kilobase RNA species from rat brain, muscle, and heart. Sucrose-gradient fractionation of rat brain poly(A) RNA yielded a high molecular weight fraction containing this mRNA, which resulted in functional Na channels when injected into oocytes. This fraction contained undetectable amounts of low molecular weight RNA. The high molecular weight Na channel RNA was selected from rat brain poly(A) RNA by hybridization to a single-strand antisense cDNA clone. Translation of this RNA in Xenopus oocytes resulted in the appearance of tetrodotoxin-sensitive voltage-sensitive Na channels in the oocyte membrane. These results demonstrate that mRNA encoding the alpha subunit of the rat brain Na channel, in the absence of any beta-subunit mRNA, is sufficient for translation to give functional channels in oocytes.", "date": "1986-10-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "83", "number": "19", "publisher": "National Academy of Sciences", "pagerange": "7503-7507", "id_number": "CaltechAUTHORS:GOLpnas86", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:GOLpnas86", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "Medical Research Council of Canada" }, { "agency": "American Heart Association" }, { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)" }, { "agency": "National Multiple Sclerosis Society" } ] }, "pmcid": "PMC386747", "primary_object": { "basename": "GOLpnas86.pdf", "url": "https://authors.library.caltech.edu/records/e1krd-76h54/files/GOLpnas86.pdf" }, "resource_type": "article", "pub_year": "1986", "author_list": "Goldin, Alan L.; Snutch, Terry P.; et el." }, { "id": "https://authors.library.caltech.edu/records/70sjs-3ja37", "eprint_id": 105994, "eprint_status": "archive", "datestamp": "2023-08-19 18:29:20", "lastmod": "2023-10-20 22:57:44", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Snutch-T-P", "name": { "family": "Snutch", "given": "Terry P." } }, { "id": "L\u00fcbbert-H", "name": { "family": "L\u00fcbbert", "given": "Hermann" } }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Expression and modulation of voltage-gated calcium channels after RNA injection in Xenopus oocytes", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1986 American Association for the Advancement of Science. \n\n13 August 1985; accepted 17 December 1985. \n\nWe thank J. P. Leonard, A. M. Gurney, J. M. Nerbonne, J. Nargeot, and D. Fambrough for helpful discussion. N.D. was a Myron R. Bantrell Fellow, T.P.S. was a postdoctoral fellow of the American Heart Association/Greater Los Angeles Affiliate, and H.L. was a fellow of the Deutsche Forschungsgemeinschaft. Supported by NIH grants GM-10991 and GM-29836.", "abstract": "Calcium ions flow into cells through several distinct classes of voltage-dependent calcium-selective channels. Such fluxes play important roles in electrical signaling at the cell membrane and in chemical signaling within cells. Further information about calcium channels was obtained by injecting RNA isolated from rat brain, heart and skeletal muscle into Xenopus oocytes. Macroscopic currents through voltage-operated calcium channels were resolved when the endogenous calcium-dependent chloride current was blocked by replacing external calcium with barium and chloride with methanesulfonate. The resulting barium current was insensitive to tetrodotoxin but was completely blocked by cadmium or cobalt. With both heart and brain RNA at least two distinct types of calcium ion conductance were found, distinguishable by their time course and inactivation properties. In oocytes injected with heart RNA, the slowly inactivating component was selectively blocked by the calcium-channel antagonist nifedipine. Barium ion currents induced by heart RNA were modulated by isoproterenol, cyclic adenosine monophosphate, and acetylcholine.", "date": "1986-03-07", "date_type": "published", "publication": "Science", "volume": "231", "number": "4742", "publisher": "American Association for the Advancement of Science", "pagerange": "1147-1150", "id_number": "CaltechAUTHORS:20201012-152226204", "issn": "0036-8075", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-152226204", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Natural Sciences and Engineering Research Council of Canada (NSERC)" }, { "agency": "American Heart Association, Greater Los Angeles Affiliate" }, { "agency": "Deutsche Forschungsgemeinschaft (DFG)" }, { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "GM-29836" } ] }, "doi": "10.1126/science.2418503", "resource_type": "article", "pub_year": "1986", "author_list": "Dascal, Nathan; Snutch, Terry P.; et el." }, { "id": "https://authors.library.caltech.edu/records/t5hns-ca377", "eprint_id": 105995, "eprint_status": "archive", "datestamp": "2023-08-19 18:25:06", "lastmod": "2023-10-20 22:57:53", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Chabala-L-D", "name": { "family": "Chabala", "given": "Lee D." } }, { "id": "Gurney-A-M", "name": { "family": "Gurney", "given": "Alison M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Dose-response of acetylcholine receptor channels opened by a flash-activated agonist in voltage-clamped rat myoballs", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1986 The Physiological Society. \n\n(Received 29 January 1985) \n\nThe authors would like to thank Drs R. Horn and D. P. Corey for suggestions regarding the voltage-clamp techniques used, Dr M. E. Krouse for helpful discussions, and Dr R. E. Sheridan for help in setting up the equipment. Drs B. F. Erlanger and N. H. Wassermann provided the Bis-Q crystals, and the cultured cells were maintained by T. Stevens. This work was supported by fellowships from the Muscular Dystrophy Association of America (L. D. C.) and the Del E. Webb Foundation (L. D. C. & A. M. G.), a Fulbright-Hayes Travel Grant (A. M. G.), and by U.S.P.H.S. grant No. NS-11756.", "abstract": "Whole\u2010cell or single\u2010channel currents through acetylcholine (ACh) receptor channels were studied in voltage\u2010clamped rat myoballs or in excised membrane patches from myoballs. The recording pipette contained CsCl to suppress outward currents, and tetrodotoxin was used to help suppress Na+ currents. To minimize problems associated with bath applied agonists, myoballs were bathed in a solution containing the inactive (cis) isomer of the photo\u2010isomerizable azobenzene derivative, Bis\u2010Q. Calibrated light flashes of varying intensity were presented to produce concentration jumps of agonist, trans\u2010Bis\u2010Q. The resulting whole\u2010cell current relaxations through ACh channels approach a steady state along an exponential time course, then decline as the newly created agonist diffuses away over the next few seconds. The dose\u2010response relationship was inferred from Hill (double\u2010log) plots for myoballs bathed in 500 nM\u2010cis\u2010Bis\u2010Q at three membrane potentials. At low agonist concentrations (less than 300 nM\u2010trans\u2010Bis\u2010Q), the slope of the Hill plot averaged 1.62 at \u2010150 mV, 1.89 at \u2010100 mV, and 2.05 at +80 mV. These results are consistent with an apparent agonist affinity constant that decreases with membrane depolarization and shifts the responses further down on the dose\u2010response curve. When the myoballs were bathed in higher concentrations of cis\u2010Bis\u2010Q (1.5\u201020 microM), the slope of the Hill plot was reduced at all membrane potentials, although it was still closer to two at positive potentials. This is expected from the known sigmoid shape of the dose\u2010response relation. The shallow dependence of the Hill slope on agonist concentration suggests the presence of negative cooperativity in the over\u2010all binding of agonist molecules. Following treatment of the membrane with dithiothreitol to reduce disulphide groups, the Hill slope for the reversibly bound agonist, trans\u2010Bis\u2010Q, remained near two. The kinetics of currents at hyperpolarized membrane potentials became complicated at higher agonist concentrations in a manner that was consistent with open\u2010channel block by trans\u2010Bis\u2010Q; the currents showed a slow secondary increase in conductance. Averaged single\u2010channel recordings at higher agonist concentrations resemble macroscopic relaxations under comparable conditions. Furthermore, those recordings also suggested that open channels are blocked by trans\u2010Bis\u2010Q at concentrations greater than 2 microM; the block depends strongly on membrane potential and increases with hyperpolarization. Currents at positive membrane potentials showed no evidence of open\u2010channel block.", "date": "1986-02-01", "date_type": "published", "publication": "Journal of Physiology", "volume": "371", "number": "1", "publisher": "Physiological Society", "pagerange": "407-433", "id_number": "CaltechAUTHORS:20201012-152226305", "issn": "0022-3751", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-152226305", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Muscular Dystrophy Association of America" }, { "agency": "Del E. Webb Foundation" }, { "agency": "Fulbright-Hayes" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1113/jphysiol.1986.sp015983", "pmcid": "PMC1192732", "resource_type": "article", "pub_year": "1986", "author_list": "Chabala, Lee D.; Gurney, Alison M.; et el." }, { "id": "https://authors.library.caltech.edu/records/x4w9g-4f993", "eprint_id": 9136, "eprint_status": "archive", "datestamp": "2023-08-22 04:29:44", "lastmod": "2023-10-23 16:13:07", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Dascal-N", "name": { "family": "Dascal", "given": "Nathan" } }, { "id": "Lotan-I", "name": { "family": "Lotan", "given": "Ilana" } }, { "id": "Gillo-B", "name": { "family": "Gillo", "given": "Boaz" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Lass-Y", "name": { "family": "Lass", "given": "Yoram" } } ] }, "title": "Acetylcholine and phorbol esters inhibit potassium currents evoked by adenosine and cAMP in Xenopus oocytes", "ispublished": "pub", "full_text_status": "public", "keywords": "muscarinic response; purinergic response; protein kinase C; second messengers; neurotransmitter interactions", "note": "\u00a9 1985 by the National Academy of Sciences. \n\nCommunicated by Howard C. Berg, April 22, 1985. \n\nWe are grateful to Prof. E.M. Landau and Dr. Alison M. Gurney for the helpful discussion. This work was supported in part by grants from Recanati and Schreiber Foundations and by National Institutes of Health Grant GM29836. N.D. is a recipient of the Bantrell and Fulbright Postdoctoral Fellowships. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - DASpnas85.pdf
", "abstract": "In Xenopus laevis oocytes, adenosine and other purinergic agonists induce a K+-conductance increase that is fully mimicked by intracellular application of cAMP. Acetylcholine suppresses the K+-conductance increase caused by adenosine, by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, or by intracellular injection of cAMP. This effect of acetylcholine is not mimicked by intracellular injection of Ca2+ or of the Ca-mobilizing agent inositol 1,4,5-trisphosphate. However, adenosine and cAMP responses are inhibited by 4\u00df -phorbol 12,13-dibutyrate and 4\u00df -phorbol 12-myristate 13-acetate. These results suggest that, in Xenopus oocytes, the muscarinic inhibition of purinergic and cAMP responses is mediated through the activation of the phospholipid-dependent, Ca-activated protein kinase (protein kinase C).", "date": "1985-09-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "82", "number": "17", "publisher": "National Academy of Sciences", "pagerange": "6001-6005", "id_number": "CaltechAUTHORS:DASpnas85", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:DASpnas85", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "pmcid": "PMC390682", "primary_object": { "basename": "DASpnas85.pdf", "url": "https://authors.library.caltech.edu/records/x4w9g-4f993/files/DASpnas85.pdf" }, "resource_type": "article", "pub_year": "1985", "author_list": "Dascal, Nathan; Lotan, Ilana; et el." }, { "id": "https://authors.library.caltech.edu/records/0ee6t-n7480", "eprint_id": 32181, "eprint_status": "archive", "datestamp": "2023-08-19 17:58:21", "lastmod": "2023-10-17 23:01:21", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Gurney-A-M", "name": { "family": "Gurney", "given": "Alison M." } }, { "id": "Nerbonne-J-M", "name": { "family": "Nerbonne", "given": "Jeanne M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Photoinduced Removal of Nifedipine Reveals Mechanisms of Calcium Antagonist Action on Single Heart Cells", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1985 The Rockefeller University Press. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. \n\nOriginal version received 4 December 1984 and accepted version received 7 May 1985. \n\nThis work was supported by the Del E. Webb Foundation (fellowship to A.M.G.), Fulbright Hayes (travel grant to A.M.G.), the Los Angeles Affiliate of the American Heart Association (fellowships to A.M.G. and J.M.N.), the National Office of the American Heart Association (established investigatorship to J.M.N.), and the National Institutes of Health (grant GM-29836).\n\nPublished - GURjgp85.pdf
", "abstract": "The currents through voltage-activated calcium channels in heart cell membranes are suppressed by dihydropyridine calcium antagonists such as nifedipine. Nifedipine is photolabile, and the reduction of current amplitude by this drug can be reversed within a few milliseconds after a 1-ms light flash. The blockade by nifedipine and its removal by flashes were studied in isolated myocytes from neonatal rat heart using the whole-cell clamp method. The results suggest that nifedipine interacts with closed, open, and inactivated calcium channels. It is likely that at the normal resting potential of cardiac cells, the suppression of current amplitude arises because nifedipine binds to and stabilizes channels in the resting, closed state. Inhibition is enhanced at depolarized membrane potentials, where interaction with inactivated channels may also become important. Additional block of open channels is suggested when currents are carried by Ba^(2+) but is not indicated with Ca^(2+) currents. Numerical simulations reproduce the experimental observations with molecular dissociation constants on the order of 10^(-7) M for closed and open channels and 10^(-8) M for inactivated channels.", "date": "1985-09", "date_type": "published", "publication": "Journal of General Physiology", "volume": "86", "number": "3", "publisher": "Rockefeller University Press", "pagerange": "353-379", "id_number": "CaltechAUTHORS:20120629-070830645", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120629-070830645", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Del E. Webb Foundation" }, { "agency": "Fullbright Hayes" }, { "agency": "American Heart Association, Los Angeles Affiliate" }, { "agency": "American Heart Association, National Office" }, { "agency": "NIH", "grant_number": "GM-29836" } ] }, "doi": "10.1085/jgp.86.3.353", "pmcid": "PMC2228798", "primary_object": { "basename": "GURjgp85.pdf", "url": "https://authors.library.caltech.edu/records/0ee6t-n7480/files/GURjgp85.pdf" }, "resource_type": "article", "pub_year": "1985", "author_list": "Gurney, Alison M.; Nerbonne, Jeanne M.; et el." }, { "id": "https://authors.library.caltech.edu/records/bs842-hf472", "eprint_id": 32166, "eprint_status": "archive", "datestamp": "2023-08-19 17:56:58", "lastmod": "2023-10-17 23:00:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Krouse-M-E", "name": { "family": "Krouse", "given": "Mauri E." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Wassermann-N-H", "name": { "family": "Wassermann", "given": "Norbert H." } }, { "id": "Erlanger-B-F", "name": { "family": "Erlanger", "given": "Bernard F." } } ] }, "title": "Rates and Equilibria for a Photoisomerizable Antagonist at the Acetylcholine Receptor of Electrophorus Electroplaques", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1985 The Rockefeller University Press. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. \n\nOriginal version received 24 September 1984 and accepted version received 29 March 1985. \n\nWe thank R. Spencer for assistance in all phases of the project and J. M. Nerbonne, J. Pine, and D. Van Essen for advice. This research was sponsored by the National Institutes of Health (research grants NS-11756 and NS-15581) and by a grant-in-aid from the Muscular Dystrophy Association of America. M.E.K. was supported in part by predoctoral training grants from the National Institutes of\nHealth (GM-07737) and by the Weigle Memorial Fund.\n\nPublished - KROjgp85.pdf
", "abstract": "Voltage-jump and light-flash experiments have been performed on isolated Electrophorus electroplaques exposed simultaneously to nicotinic agonists and to the photoisomerizable compound 2,2'-bis-[\u03b1-(trimethylammonium)methyl]-azobenzene (2BQ). Dose-response curves are shifted to the right in a nearly parallel fashion by 2BQ, which suggests competitive antagonism; dose-ratio analyses show apparent dissociation constants of 0.3 and 1 \u00b5M for the cis and trans isomers, respectively. Flash-induced trans \u2192 cis concentration jumps produce the expected decrease in agonist-induced conductance; the time constant is several tens of milliseconds. From the concentration dependence of these rates, we conclude that the association and dissociation rate constants for the cis-2BQ-receptor binding are approximately ~ 10^8 M^(-1) s^(-1) and 60 s^(-1) at 20\u00baC; the Q_(10) is 3. Flash-induced cis \u2192 trans photoisomerizations produce molecular rearrangements of the ligand-receptor complex, but the resulting relaxations probably reflect the kinetics of buffered diffusion rather than of the interaction between trans-2BQ and the receptor. Antagonists seem to bind about an order of magnitude more slowly than agonists at nicotinic receptors.", "date": "1985-08-01", "date_type": "published", "publication": "Journal of General Physiology", "volume": "86", "number": "2", "publisher": "Rockefeller University Press", "pagerange": "235-256", "id_number": "CaltechAUTHORS:20120628-075949869", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120628-075949869", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-15581" }, { "agency": "Muscular Dystrophy Association of America" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM-07737" }, { "agency": "Weigle Memorial Fund" } ] }, "doi": "10.1085/jgp.86.2.235", "pmcid": "PMC2228782", "primary_object": { "basename": "KROjgp85.pdf", "url": "https://authors.library.caltech.edu/records/bs842-hf472/files/KROjgp85.pdf" }, "resource_type": "article", "pub_year": "1985", "author_list": "Krouse, Mauri E.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/ky1qf-ga910", "eprint_id": 105996, "eprint_status": "archive", "datestamp": "2023-08-19 17:56:51", "lastmod": "2023-10-20 22:58:00", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Chabala-L-D", "name": { "family": "Chabala", "given": "Lee D." } }, { "id": "Gurney-A-M", "name": { "family": "Gurney", "given": "Alison M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Photoactivation and dissociation of agonist molecules at the nicotinic acetylcholine receptor in voltage-clamped rat myoballs", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1985 The Biophysical Society. Published by Elsevier Under an Elsevier user license. \n\nReceived for publication 5 November 1984 and in final form 8 April 1985. \n\nThe authors would like to thank Drs. B. Sakmann, D. Corey, and R. Horn for help with techniques and Dr. R. E. Sheridan for help in setting up the equipment. The Bis-Q crystals were kindly provided by Drs. B. F. Erlanger and N. H. Wasserman; Ms. T. Stevens maintained the cultured cells. \n\nThis work was supported by fellowships from the Muscular Dystrophy Association of America (L. D. Chabala) and the Del E. Webb Foundation (L. D. Chabala and A. M. Gurney), a Fulbright-Hayes travel grant (A. M. Gurney) and by United States Public Health Service grant No. NS-11756.", "abstract": "The photochemical properties of the azobenzene derivative, Bis-Q, were exploited to carry out an agonist concentration jump followed by a molecular rearrangement of bound agonist molecules at acetylcholine (ACh) receptor channels of voltage-clamped rat myoballs. Myoballs were bathed in solutions containing low concentrations of cis-Bis-Q, the inactive isomer. Whole-cell current relaxations were studied following a light flash that produced a concentration jump of agonist, trans-Bis-Q, followed by a second flash that produced net trans----cis photoisomerizations of Bis-Q molecules. The concentration-jump relaxation provided a measure of the mean burst duration for ACh receptor channels occupied by trans-Bis-Q (7.7 ms, 22 degrees C). The second current relaxation was a more rapid conductance decrease (phase 1, \u03c4 = 0.8 ms). Phase 1 may represent either the burst duration for receptors initially occupied by a single cis- and a single trans-Bis-Q molecule or that for unliganded receptors. Single-channel current recordings from excised outside-out membrane patches showed that single channels open following an agonist concentration jump comparable to that used in the whole-cell experiments; when many such records were averaged, a synthetic macroscopic relaxation was produced. Individual open channels closed faster following a flash that promoted trans\u2192cis photoisomerizations of the bound ligand, thus confirming the whole-cell observations of phase 1.", "date": "1985-08", "date_type": "published", "publication": "Biophysical Journal", "volume": "48", "number": "2", "publisher": "Biophysical Society", "pagerange": "241-246", "id_number": "CaltechAUTHORS:20201012-152226405", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-152226405", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Muscular Dystrophy Association of America" }, { "agency": "Del E. Webb Foundation" }, { "agency": "Fulbright-Hayes" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1016/s0006-3495(85)83777-2", "pmcid": "PMC1329315", "resource_type": "article", "pub_year": "1985", "author_list": "Chabala, Lee D.; Gurney, Alison M.; et el." }, { "id": "https://authors.library.caltech.edu/records/te5qt-8e375", "eprint_id": 6169, "eprint_status": "archive", "datestamp": "2023-08-22 04:27:03", "lastmod": "2023-10-23 16:13:48", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "White-M-M", "name": { "family": "White", "given": "Michael M." } }, { "id": "Mayne-K-M", "name": { "family": "Mayne", "given": "Katharine Mixter" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Davidson-N", "name": { "family": "Davidson", "given": "Norman" } } ] }, "title": "Mouse-Torpedo Hybrid Acetylcholine Receptors: Functional Homology Does not Equal Sequence Homology", "ispublished": "pub", "full_text_status": "public", "keywords": "Xenopus oocytes; SP6 RNA polymerase; cDNA clones; Ion channels; in ovo translation", "note": "\u00a9 1985 by the National Academy of Sciences \n\nCommunicated by Norman Davidson, April 5, 1985 \n\nWe thank Drs. T. Claudio, S. Heinemann, and D. Noonan for kindly providing Torpedo AcChoR cDNA clones, Dr. D. Melton for providing plasmid pSP62-PL and information concerning the SP6 transcription system, Drs. P. Sharp and M. M. Konarska for information concerning the use of GpppG in the transcription reaction, and Dr. N. Dascal for helpful suggestions on the handling of oocytes. We appreciate the counsel of Dr. J.H. Richards about the relation (or lack thereof) of primary sequence to functional domains in proteins. We are especially grateful to Dr. T. Steitz for suggesting the use of helical net projections in the search for significant homologies. This work has been supported in part by research grants GM-10991 (N.D.) and NS-11756 (H.A.L.) from the National Institutes of Health, a research grant from the Muscular Dystrophy Association to N.D., postdoctoral fellowships from the Procter and Gamble Co. and the Muscular Dystrophy Association to M.M.W., and Predoctoral Training Grant GM-07616 from the National Institutes of Health to K.M.M. \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U.S.C. \u00a71734 solely to indicate this fact.\n\nPublished - WHIpnas85.pdf
", "abstract": "The nicotinic acetylcholine (AcCho) receptor (AcChoR) is a multisubunit protein complex of stoichiometry alpha 2\u00df gamma delta . The several subunits show homology with each other within a given species; in addition, homology is found between analogous subunits between species. We have used the phage SP6 RNA polymerase transcription system to produce single-species RNA in vitro for various AcChoR subunits from cDNAs. Injection of an equimolar mixture of RNA for the alpha, \u00df, gamma, and delta subunits of Torpedo californica AcChoR into Xenopus oocytes results in the appearance of functional receptors in the oocyte membrane. No response to AcCho is detected when the \u00df or gamma subunit RNA is omitted, and a small response is seen when the delta subunit RNA is omitted. Replacement of Torpedo delta subunit RNA by the mouse BC3H-1 cell line AcChoR delta subunit RNA leads to the formation of functional receptors that show a 3-4-fold greater response to AcCho than does the full Torpedo complex. No response is seen when the mouse delta RNA replaces Torpedo gamma RNA. By amino acid homology profile comparisons, the mouse delta subunit appears to be moderately but not highly similar to the Torpedo delta subunit; the apparent similarity to the Torpedo gamma subunit is only slightly less. Therefore, the features of the primary sequence that determine the functional delta character of the mouse polypeptide are not revealed by simple homology comparisons.", "date": "1985-07-15", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "82", "number": "14", "publisher": "National Academy of Sciences", "pagerange": "4852-4856", "id_number": "CaltechAUTHORS:WHIpnas85", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:WHIpnas85", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-10991" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Procter and Gamble" }, { "agency": "NIH Predoctoral Fellowship", "grant_number": "GM-07616" } ] }, "pmcid": "PMC391003", "primary_object": { "basename": "WHIpnas85.pdf", "url": "https://authors.library.caltech.edu/records/te5qt-8e375/files/WHIpnas85.pdf" }, "resource_type": "article", "pub_year": "1985", "author_list": "White, Michael M.; Mayne, Katharine Mixter; et el." }, { "id": "https://authors.library.caltech.edu/records/sj37a-kd097", "eprint_id": 106158, "eprint_status": "archive", "datestamp": "2023-08-22 04:18:26", "lastmod": "2023-10-20 23:09:01", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Richard-Sylvain", "name": { "family": "Richard", "given": "Sylvain" } }, { "id": "Nerbonne-J-M", "name": { "family": "Nerbonne", "given": "Jeanne M." } }, { "id": "Nargeot-J", "name": { "family": "Nargeot", "given": "Joel" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Garnier-D", "name": { "family": "Garnier", "given": "Didier" } } ] }, "title": "Photochemically produced intracellular concentration jumps of cAMP mimic the effects of catecholamines on excitation-contraction coupling in frog atrial fibers", "ispublished": "pub", "full_text_status": "public", "keywords": "cAMP; Slow inward current; Phasic tension; Tonic tension; K outward currents", "note": "\u00a9 1985 Springer. \n\nReceived 04 July 1984; Accepted 03 December 1984.", "abstract": "Previously, we reported that concentration jumps of cAMP produced by light flashes in the presence of a photosensitive analogue of cAMP increase the amplitude of the slow inward current (I_(si)) in isolated bullfrog atrial trabeculae (Nargeot et al. 1983). Here, using newly designed photolabile cyclic nucleotides (Nerbonne et al. 1984a), we have examined the effects of intracellular concentration jumps of cAMP and cGMP on excitation-contraction coupling in frog heart. Concentration jumps of cAMP increase the amplitude and the duration of action potentials, increase I_(si) and twitch tension. Following single flashes, maximum responses are observed in 10\u201330 s and recovery times are 30\u2013120 s. The time courses of the cAMP-induced increases in I_(si) and phasic tension amplitudes are parallel, implying a direct correlation between Ca\u00b2\u207a influx through the slow channels and the development of phasic tension. Although the amplitudes are increased severalfold, cAMP jumps do not measurably alter the kinetics or voltage dependences of the current or tension. cAMP concentration jumps increase the delayed K\u207a current (I_K) and decrease tonic tension; relaxation of contraction is not, however, influenced by cAMP jumps. Concentration jumps of cGMP, on the other hand, have no measurable effects on the action potential, I_(si), I_K or tension in this preparation.", "date": "1985-03", "date_type": "published", "publication": "Pfl\u00fcgers Archiv European Journal of Physiology", "volume": "403", "number": "3", "publisher": "Springer", "pagerange": "312-317", "id_number": "CaltechAUTHORS:20201020-073939931", "issn": "0031-6768", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201020-073939931", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1007/bf00583606", "resource_type": "article", "pub_year": "1985", "author_list": "Richard, Sylvain; Nerbonne, Jeanne M.; et el." }, { "id": "https://authors.library.caltech.edu/records/7j2re-zv872", "eprint_id": 105997, "eprint_status": "archive", "datestamp": "2023-08-22 04:16:23", "lastmod": "2023-10-20 22:58:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Kegel-D-R", "name": { "family": "Kegel", "given": "Daniel R." } }, { "id": "Wolf-B-D", "name": { "family": "Wolf", "given": "Benjamin D." } }, { "id": "Sheridan-R-E", "name": { "family": "Sheridan", "given": "Robert E." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Software for electrophysiological experiments with a personal computer", "ispublished": "pub", "full_text_status": "restricted", "keywords": "voltage-clamp; single channels; BASIC; IBM personal computer", "note": "\u00a9 1985 Published by Elsevier. \n\nReceived 17 October 1984, Revised 22 November 1984, Accepted 23 November 1984. \n\nWe thank Mr. Michael Walsh in Caltech's Biology Electronics Shop for designing and building the window discriminator and the timeshare circuit. We thank Alison M. Gurney, Jeanne M. Nerbonne, Lee D. Chabala, Alan Finkel, Michael M. White, and Jerome Pine for testing and advising on the system. This research was sponsored by the National Institutes of Health (NS-11756).", "abstract": "These programs were written for the IBM personal computer equipped with a Tecmar Labmaster analog interface board. The software operates under the MS-DOS operating system; it is written in compiled BASIC and employs short machine-language subroutines for crucial functions. Details are presented on the analog interface routines which make special use of the 9513 counter/timer chip on the Labmaster. Time resolution is about 32 \u03bcs per sample on the PC and 15 \u03bcs on the PC/AT. One series of programs performs traditional voltage- and current-clamp experiments on macroscopic currents in whole cells. A second series interfaces with a hardware window discriminator to capture single-channel events in a recirculating buffer, a stimulus can be triggered to fire during an open channel. Off-line programs perform standard analyses, and further processing can be performed with standard spreadsheet programs.", "date": "1985-02", "date_type": "published", "publication": "Journal of Neuroscience Methods", "volume": "12", "number": "4", "publisher": "Elsevier", "pagerange": "317-330", "id_number": "CaltechAUTHORS:20201012-152226500", "issn": "0165-0270", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-152226500", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1016/0165-0270(85)90016-0", "resource_type": "article", "pub_year": "1985", "author_list": "Kegel, Daniel R.; Wolf, Benjamin D.; et el." }, { "id": "https://authors.library.caltech.edu/records/jbg5e-cgt08", "eprint_id": 106000, "eprint_status": "archive", "datestamp": "2023-08-19 17:03:01", "lastmod": "2023-10-20 22:58:25", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nerbonne-J-M", "name": { "family": "Nerbonne", "given": "Jeanne M." } }, { "id": "Richard-Sylvain", "name": { "family": "Richard", "given": "Sylvain" } }, { "id": "Nargeot-J", "name": { "family": "Nargeot", "given": "Joel" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "New photoactivatable cyclic nucleotides produce intracellular jumps in cyclic AMP and cyclic GMP concentrations", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1984 Nature Publishing Group. \n\nReceived 31 January; accepted 12 April 1984. \n\nWe thank Roger Spencer for assistance with the HPLC experiments and David Trentham for helpful discussions. This research was supported by the NIH (grant GM-29836), by INSERM, France (grant 835012) and by the American Heart Association (Postdoctoral Fellowship to J.M.N.). We also thank Centre d'etudes et de Recherches Servier, Neuilly/Seine\n(France) for a travel grant to J.M.N.", "abstract": "The cyclic nucleotides cyclic AMP and cyclic GMP are important intracellular messengers mediating the responses to neurotransmitters and neurohormones and regulating cellular function over a wide range of time scales. Despite the widespread acceptance of this second messenger mechanism in many systems, much remains unknown about their mechanism of action, except that such events are associated with increases or decreases in intracellular cyclic nucleotides. Quantitative descriptions of cyclic nucleotide-dependent processes are hampered by the absence of a means by which intracellular cyclic nucleotide concentrations can be accurately controlled. We have now designed, synthesized and characterized new, substituted photolabile cyclic nucleotide analogues, the 4,5-dimethoxy-2-nitrobenzyl esters of cyclic AMP and cyclic GMP (Fig. 1), which are physiologically inert before irradiation and which liberate free cyclic AMP or cyclic GMP on absorption of a photon. The thermal properties and photolysis rates and efficiencies of light-induced release of cyclic nucleotides from these analogues are more favourable than for the simple o-nitrobenzyl derivatives used previously. These molecules should permit intracellular 'concentration jumps' of cyclic AMP or cyclic GMP to be produced in cells under physiological investigation with spatial and temporal resolution unmatched by conventional techniques.", "date": "1984-07-05", "date_type": "published", "publication": "Nature", "volume": "310", "number": "5972", "publisher": "Nature Publishing Group", "pagerange": "74-76", "id_number": "CaltechAUTHORS:20201012-160322096", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-160322096", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "Institut national de la sant\u00e9 et de la recherche m\u00e9dicale (INSERM)", "grant_number": "835012" }, { "agency": "American Heart Association" }, { "agency": "Laboratoire Servier" } ] }, "doi": "10.1038/310074a0", "resource_type": "article", "pub_year": "1984", "author_list": "Nerbonne, Jeanne M.; Richard, Sylvain; et el." }, { "id": "https://authors.library.caltech.edu/records/9wr09-zsm83", "eprint_id": 5789, "eprint_status": "archive", "datestamp": "2023-08-22 03:28:01", "lastmod": "2023-10-23 16:12:05", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nargeot-J", "name": { "family": "Nargeot", "given": "Joel" } }, { "id": "Nerbonne-J-M", "name": { "family": "Nerbonne", "given": "Jeanne M." } }, { "id": "Engels-J", "name": { "family": "Engels", "given": "Joachim" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Time Course of the Increase in the Myocardial Slow Inward Current after a Photochemically Generated Concentration Jump of Intracellular cAMP", "ispublished": "pub", "full_text_status": "public", "keywords": "calcium channel; voltage clamp; o-nitrobenzyl ester; cGMP; \u00df-adrenergic agonist", "note": "\u00a9 1983 by the National Academy of Sciences \n\nCommunicated by James Bonner, January 17, 1983 \n\nWe thank Roger Spencer for assistance in all phases of the experiments and G. N. Gill, F. Strumwasser, D. Trentham, and R. Y. Tsien for helpful discussion. This research was supported by the National Institutes of Health (Research Career Development Award NS-272 to H.A.L. and Grant GM-29836), by the American Heart Association (a Postdoctoral Fellowship to J.M.N.), and by the Fulbright Exchange Program (a Scholarship to J.N.). \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U. S. C. \u00a71734 solely to indicate this fact\n\nPublished - NARpnas83.pdf
", "abstract": "Voltage-clamped atrial trabeculae from bullfrog hearts were exposed to membrane-permeant photolyzable o-nitrobenzyl esters of cAMP and cGMP. UV flashes produced intracellular concentration jumps of cAMP or cGMP. With the cAMP derivative, flashes resulted in an increased slow inward current (Isi), producing a broadened action potential. The Isi reached a maximum 10-30 sec after the flash and decreased over the next 60-300 sec. The first increases were observable within 150 msec; this value is an upper limit imposed by the instrumentation. Responses to flashes lasted longer at higher drug concentrations and in the presence of the phosphodiesterase inhibitor papaverine; effects of flashes developed and decreased faster at higher temperature. Although the amplitude of the Isi was increased, its waveform and voltage sensitivity were not affected. Intracellular concentration jumps of cAMP failed to affect the muscarinic K+ conductance. There were no observable effects of cGMP concentration jumps. The data confirm (i) that cAMP regulates the Isi and (ii) that the 5- to 10-sec delay between application of \u00df-agonists and the onset of positive inotropic effects, observed in previous studies, has been correctly ascribed to events prior to the interaction between cAMP and protein kinase.", "date": "1983-04-15", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "80", "number": "8", "publisher": "National Academy of Sciences", "pagerange": "2395-2399", "id_number": "CaltechAUTHORS:NARpnas83", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:NARpnas83", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "American Heart Association" }, { "agency": "Fulbright Exchange Program" } ] }, "pmcid": "PMC393827", "primary_object": { "basename": "NARpnas83.pdf", "url": "https://authors.library.caltech.edu/records/9wr09-zsm83/files/NARpnas83.pdf" }, "resource_type": "article", "pub_year": "1983", "author_list": "Nargeot, Joel; Nerbonne, Jeanne M.; et el." }, { "id": "https://authors.library.caltech.edu/records/3hwe4-bs963", "eprint_id": 105999, "eprint_status": "archive", "datestamp": "2023-08-19 15:51:39", "lastmod": "2023-10-20 22:58:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nerbonne-J-M", "name": { "family": "Nerbonne", "given": "Jeanne M." } }, { "id": "Sheridan-R-E", "name": { "family": "Sheridan", "given": "Robert E." } }, { "id": "Chabala-L-D", "name": { "family": "Chabala", "given": "Lee D." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "cis-3,3'-Bis-[\u03b1-(trimethylammonium)methyl]azobenzene (cis-Bis-Q). Purification and properties at acetylcholine receptors of Electrophorus electroplaques", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1983 American Society for Pharmacology and Experimental Therapeutics. \n\nWe thank Roger Spencer for assistance in all phases of the project, Mauri Krouse for performing some of the electrophysiological measurements, and N. H. Wassermann and B. F. Erlanger for furnishing Bis-Q. \n\nThis work was supported by National Institutes of Health Grant NS-11756. \n\nRecipient of a Fellowship from the American Heart Association. \n\nRecipient of a Fellowship from the Del E. Webb Foundation. \n\nRecipient of Research Career Development Award NS-272 from the National Institutes of Health.", "abstract": "The cis and trans isomers of the photoisomerizable compound, 3,3'-bis-[\u03b1-(trimethylammonium)methyl]azobenzene (Bis-Q), were purified by high-performance liquid chromatography using the ion-pair partitioning technique on a reverse-phase column. Solutions of cis-Bis-Q are stable at -20 degrees; at 25 degrees, thermal isomerization proceeds at a rate of 0.65%/day. cis-Bis-Q is less than 1% as potent a nicotinic agonist as the trans configuration. At concentrations of 1.5 microM or less, cis-Bis-Q exerts little or no blockade of the conductances induced by agonists. In voltage-clamped Electrophorus electroplaques exposed to cis-Bis-Q, laser flashes induce cis leads to trans photoisomerizations and increase the agonist-induced current by a factor of 20 within a few milliseconds.", "date": "1983-03", "date_type": "published", "publication": "Molecular pharmacology", "volume": "23", "number": "2", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "344-349", "id_number": "CaltechAUTHORS:20201012-160244665", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-160244665", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "American Heart Association" }, { "agency": "Del E. Webb Foundation" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" } ] }, "resource_type": "article", "pub_year": "1983", "author_list": "Nerbonne, Jeanne M.; Sheridan, Robert E.; et el." }, { "id": "https://authors.library.caltech.edu/records/98wts-91845", "eprint_id": 106159, "eprint_status": "archive", "datestamp": "2023-08-19 15:29:34", "lastmod": "2023-10-20 23:09:05", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Braun-Sergei", "name": { "family": "Braun", "given": "Sergei" } }, { "id": "Tolkovsky-A-M", "name": { "family": "Tolkovsky", "given": "Aviva M." } }, { "id": "Steer-M-L", "name": { "family": "Steer", "given": "Michael L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Levitzki-A", "name": { "family": "Levitzki", "given": "Alexander" } } ] }, "title": "Activation and inhibition of adenylate cyclase by hormones", "ispublished": "pub", "full_text_status": "public", "keywords": "p[NH]ppG, guanosine 5\u2032-[\u03b2,\u03b3-imido]triphosphate, [S]pppG, guanosine 5\u2032-[\u03b3-thio]triphosphate, p[S]ppG, guanosine 5\u2032-[\u03b2, \u03b3-thio]triphosphate, NBF, 7-nitrobenzofurazan", "note": "\u00a9 1982 Biochemical Society.", "abstract": "Basic features of the hormone-dependent adenylate cyclases.", "date": "1982-12-01", "date_type": "published", "publication": "Biochemical Society Transactions", "volume": "10", "number": "6", "publisher": "Portland Press Ltd.", "pagerange": "496-498", "id_number": "CaltechAUTHORS:20201020-074502837", "issn": "0300-5127", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201020-074502837", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1042/bst0100496", "resource_type": "article", "pub_year": "1982", "author_list": "Braun, Sergei; Tolkovsky, Aviva M.; et el." }, { "id": "https://authors.library.caltech.edu/records/xp5dh-k7691", "eprint_id": 32516, "eprint_status": "archive", "datestamp": "2023-08-19 15:21:28", "lastmod": "2023-10-18 14:31:19", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Sheridan-R-E", "name": { "family": "Sheridan", "given": "Robert E." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Functional Stoichiometry at the Nicotinic Receptor. The Photon Cross Section for Phase 1 Corresponds to Two Bis-Q Molecules per Channel", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1982 Rockefeller University Press. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. \n\nReceived for publication 24 November 1981; in revised form 27 May 1982. \n\nWe thank S . Dunn, J. Racs, and M. A. Raftery for providing the receptor-rich membrane\nfragments from Torpedo and for performing the \u03b1-bungarotoxin binding assays . We thank N. H.\nWassermann and B. F. Erlanger for supplying the Bis-Qand QBr. We also thankM. E. Krouse, J. Nerbonne, and J. Pine for advice.\nThis research was supported by the National Institutes of Health (RCDA NS-272 to H.A .L .\nand grant NS-11756) and by a grant from the Muscular Dystrophy Associations of America.\n\nPublished - SHEjgp82.pdf
", "abstract": "These experiments examine changes in the agonist-induced conductance\nthat occur when the agonist-receptor complex is perturbed. Voltage-clamped\nElectrophorus electroplaques are exposed to the photoisomerizable agonist\ntrans-Bis-Q A 1-\u00b5s laser flash photoisomerizes some trans-Bis-Q molecules\nbound to receptors; because the cis configuration is not an agonist, receptor\nchannels close within a few hundred microseconds. This effect is called phase 1.\nWe compare (a) the fraction of channels that close during phase 1 with (b) the\nfraction of trans-Bis-Q molecules that undergo trans \u2192 cis photoisomerization.\nParameter a is measured as the fractional diminution in voltage-clamp currents\nduring phase 1. Parameter b is measured by changes in the optical spectra of\nBis-Q solutions caused by flashes . At low flash intensities, a is twice b, which\nshows that the channel can be closed by photoisomerizing either of two bound\nagonist molecules. Conventional dose-response studies with trans-Bis-Q also give\na Hill coefficient of two. As a partial control for changes in the photochemistry\ncaused by binding of Bis-Q to receptors, spectral measurements are performed\non the photoisomerizable agonist QBr, covalently bound to solubilized acetylcholine\nreceptors from Torpedo. The bound and free agonist molecules have the\nsame photoisomerization properties. These results verify the concept that the\nopen state of the acetylcholine receptor channel is much more likely to be\nassociated with the presence of two bound agonist molecules than with a single\nsuch molecule.", "date": "1982-10-01", "date_type": "published", "publication": "Journal of General Physiology", "volume": "80", "number": "4", "publisher": "Rockefeller University Press", "pagerange": "499-515", "id_number": "CaltechAUTHORS:20120717-113744311", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120717-113744311", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "RCDA NS-272" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association of America" } ] }, "doi": "10.1085/jgp.80.4.499", "pmcid": "PMC2228709", "primary_object": { "basename": "SHEjgp82.pdf", "url": "https://authors.library.caltech.edu/records/xp5dh-k7691/files/SHEjgp82.pdf" }, "resource_type": "article", "pub_year": "1982", "author_list": "Sheridan, Robert E. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/vfaz5-h6v36", "eprint_id": 32520, "eprint_status": "archive", "datestamp": "2023-08-19 15:05:29", "lastmod": "2023-10-18 14:31:37", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Nerbonne-J-M", "name": { "family": "Nerbonne", "given": "Jeanne M." } } ] }, "title": "Physiological and Pharmacological Manipulations with Light Flashes", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1982 by Annual Reviews. Research in our laboratory is supported by grants from the Muscular Dystrophy Association, the Pew Foundation, and the National Institutes of Health (NS-11756 and GM-29836 and Career Development Award\nNS-272 to H.A.L.). J.M.N. has been supported by postdoctoral fellowships from the National Institutes of Health and the American Heart Association. During the preparation of this review, H.A.L. enjoyed the hospitality of A. Levitzki and his colleagues in the Department of Biological Chemistry, Hebrew University of Jerusalem.\n\nPublished - LESarbb82.pdf
", "abstract": "In the experiments described here, a physiological measurement is made while photochemical procedures are employed to alter (a) the concentration of a ligand near membranes or proteins or (b) the structure of the\nligand-receptor complexes. Because photochemical reactions often provide the quickest way to produce such chemical perturbations, we emphasize the kinetic information that such experiments have yielded. This information requires a suitably rapid physiological measurement, usually\nan electrical or optical one. The results often complement those obtained with other kinds of kinetic investigation (iontophoretic application of drugs, stopped-flow mixing, temperature jump, etc). Pharmacological manipulations with light flashes are especially useful for biological systems\nthat cannot be flowed, for instance membranes under electro-physiological investigation or solutions at very low temperatures.", "date": "1982-06", "date_type": "published", "publication": "Annual Review of Biophysics and Bioengineering", "volume": "11", "publisher": "Annual Reviews", "pagerange": "151-175", "id_number": "CaltechAUTHORS:20120717-132820326", "issn": "0084-6589", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120717-132820326", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Muscular Dystrophy Association" }, { "agency": "Pew Foundation" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" }, { "agency": "American Heart Association" } ] }, "doi": "10.1146/annurev.bb.11.060182.001055", "primary_object": { "basename": "LESarbb82.pdf", "url": "https://authors.library.caltech.edu/records/vfaz5-h6v36/files/LESarbb82.pdf" }, "resource_type": "article", "pub_year": "1982", "author_list": "Lester, Henry A. and Nerbonne, Jeanne M." }, { "id": "https://authors.library.caltech.edu/records/ryb7t-9fv68", "eprint_id": 12039, "eprint_status": "archive", "datestamp": "2023-08-22 03:01:07", "lastmod": "2023-10-17 16:29:17", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nargeot-J", "name": { "family": "Nargeot", "given": "Joel" } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Birdsall-N-J-M", "name": { "family": "Birdsall", "given": "Nigel J. M." } }, { "id": "Stockton-J", "name": { "family": "Stockton", "given": "Jane" } }, { "id": "Wassermann-N-H", "name": { "family": "Wassermann", "given": "Norbert H." } }, { "id": "Erlanger-B-F", "name": { "family": "Erlanger", "given": "Bernard F." } } ] }, "title": "A photoisomerizable muscarinic antagonist. Studies of binding and of conductance relaxations in frog heart", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1982 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nReceived for publication 17 April 1981 and in revised form 12 September 1981. \n\nSupported by Research Career Development Award NS-272 to H.A.L. and grants NS-11756 and NS-15581 from the National Institutes of Health, a NATO fellowship to J.N., grant 78.7.2582 from the D\u00e9l\u00e9gation G\u00e9n\u00e9rale \u00e0 la Recherche Scientifique et Technique, the Pew Foundation, the Muscular Dystrophy Association, and grant PCM-77-19280 from the National Science Foundation.\n\nPublished - NARjgp82.pdf
", "abstract": "These experiments employ the photoisomerizable compound, 3,3'-bis- [alpha-(trimethylammonium)methyl]azobenzene (Bis-Q), to study the response to muscarinic agents in frog myocardium. In homogenates from the heart, trans-Bis-Q blocks the binding of [3H]-N-methylscopolamine to muscarinic receptors. In voltage-clamped atrial trabeculae, trans- Bis-Q blocks the agonist-induced potassium conductance. The equilibrium dose-response curve for carbachol is shifted to the right, suggesting competitive blockade. Both the biochemical and electrophysiological data yield a dissociation constant of 4-5 microM for trans-Bis-Q; the cis configuration is severalfold less potent as a muscarinic blocker. Voltage-clamped preparations were exposed simultaneously to carbachol and Bis-Q and were subjected to appropriately filtered flashes (less than 1 ms duration) from a xenon flashlamp. Trans leads to cis and cis leads to trans photoisomerizations cause small (less than 20%) increases and decreases, respectively, in the agonist-induced current. The relaxation follows an S-shaped time course, including an initial delay or period of zero slope. The entire waveform is described by [1 - exp(-kt)]n. At 23 degrees C, k is approximately 3 s-1 and n is 2. Neither k nor n is affected when: (a) [Bis-Q] is varied between 5 and 100 microM; (b) [carbachol] is varied between 1 and 50 microM; (c) carbachol is replaced by other agonists (muscarine, acetylcholine, or acetyl-beta-methylcholine); or (d) the voltage is varied between the normal resting potential and a depolarization of 80 mV. However, in the range of 13-30 degrees C, k increases with temperature; the Q10 is between 2 and 2.5. In the same range, n does not change significantly. Like other investigators, we conclude that the activation kinetics of the muscarinic K+ conductance are not determined by ligand-receptor binding, but rather by a subsequent sequence of two (or more) steps with a high activation energy.", "date": "1982-04", "date_type": "published", "publication": "Journal of General Physiology", "volume": "79", "number": "4", "publisher": "Rockefeller University Press", "pagerange": "657-678", "id_number": "CaltechAUTHORS:NARjgp82", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:NARjgp82", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-272" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NIH", "grant_number": "NS-15581" }, { "agency": "North Atlantic Treaty Organization (NATO)" }, { "agency": "D\u00e9l\u00e9gation G\u00e9n\u00e9rale \u00e0 la Recherche Scientifique et Technique" }, { "agency": "Pew Foundation" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "NSF", "grant_number": "PCM-77-19280" } ] }, "doi": "10.1085/jgp.79.4.657", "pmcid": "PMC2215484", "primary_object": { "basename": "NARjgp82.pdf", "url": "https://authors.library.caltech.edu/records/ryb7t-9fv68/files/NARjgp82.pdf" }, "resource_type": "article", "pub_year": "1982", "author_list": "Nargeot, Joel; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/0wesr-0mc83", "eprint_id": 106001, "eprint_status": "archive", "datestamp": "2023-08-22 03:01:40", "lastmod": "2023-10-20 22:58:28", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Steer-M-L", "name": { "family": "Steer", "given": "Michael L." } }, { "id": "Michaelson-D-M", "name": { "family": "Michaelson", "given": "Daniel M." } } ] }, "title": "ADP-Ribosylation of Membrane Proteins in Cholinergic Nerve Terminals", "ispublished": "pub", "full_text_status": "restricted", "keywords": "ADP\u2010ribosylation; Nerve; Cholinergic; Torpedo", "note": "\u00a9 1982 International Society for Neurochemistry. \n\nReceived July 13, 1981; accepted October 22, 1981. \n\nWe thank Z. Zimmer for technical assistance and A. Levitzki for advice and support. This research was supported by the United States National Institutes of Health (grant GM-26604 to A. L., and Research Career Development Awards NS-272 to H.A.L. and NS-160 to M.L.S.) and by grants from the Muscular Dystrophy Association of America to Tel Aviv University and to the California Institute of Technology.", "abstract": "Lysed Torpedo synaptosomes or washed synaptosomal membranes were incubated with [\u00b3\u00b2P]NAD\u207a and subjected to electrophoresis on SDS\u2010polyacrylamide gels. More than eight membrane proteins were ADP\u2010ribosylated. The most intensely labeled proteins were those of M_r = 62,000 and 82,000. Radiolabeling was more intense in synaptosomes than in other subcellular fractions. Cholera toxin caused ribosylation of additional synaptosomal proteins with M_r = 42,000 and (in some preparations) 49,000. Neither endogenous nor cholera toxin\u2010catalyzed ADP\u2010ribosylation required added guanyl nucleotides. Cholera toxin increased the adenylate cyclase activity of synaptosomal membranes, suggesting that the cholera toxin substrates are regulatory components of adenylate cyclase in these synaptosomes.", "date": "1982-04", "date_type": "published", "publication": "Journal of Neurochemistry", "volume": "38", "number": "4", "publisher": "Wiley", "pagerange": "1080-1086", "id_number": "CaltechAUTHORS:20201012-160322257", "issn": "0022-3042", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-160322257", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM-26604" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-160" }, { "agency": "Muscular Dystrophy Association of America" } ] }, "doi": "10.1111/j.1471-4159.1982.tb05351.x", "resource_type": "article", "pub_year": "1982", "author_list": "Lester, Henry A.; Steer, Michael L.; et el." }, { "id": "https://authors.library.caltech.edu/records/swg0n-dff58", "eprint_id": 6584, "eprint_status": "archive", "datestamp": "2023-08-22 02:57:53", "lastmod": "2023-10-23 16:59:47", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Steer-M-L", "name": { "family": "Steer", "given": "Michael L." } }, { "id": "Levitzki-A", "name": { "family": "Levitzki", "given": "Alexander" } } ] }, "title": "Prostaglandin-Stimulated GTP Hydrolysis Associated with Activation of Adenylate Cyclase in Human Platelet Membranes", "ispublished": "pub", "full_text_status": "public", "keywords": "cholera toxin; N-ethylmaleimide; epinephrine", "note": "\u00a9 1982 by the National Academy of Sciences \n\nCommunicated by John J. Hopfield, September 14, 1981 \n\nWe thank H. Arad, D. Atlas, S. Braun, D. Cassel, M. Lowe, and M. Schramm for advice and discussion. This research was supported by the U.S.-Israel Binational Science Foundation and by the National Institutes of Health (Grants GM-26604 and GM-29836 and Research Career Development Award NS-272 to H.A.L., and Research Career Development Award NS-160 to M.L.S.). \n\nThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked \"advertisement\" in accordance with 18 U. S. C. \u00a71734 solely to indicate this fact.\n\nPublished - LESpnas82.pdf
", "abstract": "In membranes purified from human blood platelets, basal guanosine triphosphate (GTP) hydrolysis is reduced by a factor of approx 6 by exposure to N-ethylmaleimide (10 mM). This decreased background enables the detection of an additional GTP hydrolysis in the presence of prostaglandin E1 (PGE1). The PGE1-stimulated GTPase has several properties correlated with PGE1-stimulated adenylate cyclase in this preparation. The two enzymes have similar dose-response relationships (half-maximal stimulation at 0.1 \u00b5M PGE1). Exposure to cholera toxin blocks the PGE1-stimulated GTPase and activates adenylate cyclase. Both enzymes are activated by submicromolar concentrations of GTP, although the Km for the GTPase is about 10 times greater than that for the adenylate cyclase. The data are discussed in relation to the hypothesis that hormone-stimulated adenylate cyclase (i) is activated as a regulatory component binds a molecule of GTP and (ii) is deactivated as this molecule is hydrolyzed.", "date": "1982-02-01", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "79", "number": "3", "publisher": "National Academy of Sciences", "pagerange": "719-723", "id_number": "CaltechAUTHORS:LESpnas82", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LESpnas82", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Binational Science Foundation (USA-Israel)" }, { "agency": "NIH", "grant_number": "GM-26604" }, { "agency": "NIH", "grant_number": "GM-29836" }, { "agency": "NIH", "grant_number": "NS-272" }, { "agency": "NIH", "grant_number": "NS-160" } ] }, "pmcid": "PMC345823", "primary_object": { "basename": "LESpnas82.pdf", "url": "https://authors.library.caltech.edu/records/swg0n-dff58/files/LESpnas82.pdf" }, "resource_type": "article", "pub_year": "1982", "author_list": "Lester, Henry A.; Steer, Michael L.; et el." }, { "id": "https://authors.library.caltech.edu/records/0hgd9-v9y31", "eprint_id": 105974, "eprint_status": "archive", "datestamp": "2023-08-19 14:46:24", "lastmod": "2023-10-20 22:56:13", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Steer-M-L", "name": { "family": "Steer", "given": "M. L." } }, { "id": "Braun-S", "name": { "family": "Braun", "given": "S." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Levitzki-A", "name": { "family": "Levitzki", "given": "A." } } ] }, "title": "Activation of adenylate cyclase from purified platelet membranes by prostaglandin E1 and its inhibition by L-epinephrine: mechanistic effects", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1982 Raven Press.", "abstract": "Activation and inhibition of adenylate cyclase in purified human platelet membranes by hormones and guanyl nucleotides was studied. The rate constant of enzyme activation (kon) was measured using the GTP analog guanylimidodiphosphate (Gpp(NH)p), and the rate constant of enzyme deactivation (koff) was determined using the guanosine diphosphate analog guanosine 5'-0-(2-thio)-diphosphate (GDP beta S). PGE1 which was found previously (15) to accelerate kon has been found to accelerate also koff, from 0.1 sec-1 (6 min-1) to 0.2 sec-1 (12 min-1). The alpha 2-adrenergic inhibitory hormone 1-epinephrine did not alter kon, whether measured in the absence or presence of GTP, and also did not alter koff, whether measured under basal or PGE1-dependent GTP-ase activity at any concentration of GTP tested. These results suggest that the alpha 2-adrenergic receptor exerts its inhibitory effect on adenylate cyclase by a mechanism which does not involve its direct interaction with the GTP regulatory protein that is associated with the stimulatory hormone. These results support the view that the inhibitor involves the interaction of the inhibitory receptor with a GTP binding unit separate from the one mediating hormonal stimulation. This regulatory unit attenuates the adenylate cyclase activity either by interacting directly with the catalytic moiety or by modulating the interaction of the stimulatory GTP regulatory protein with the catalytic moiety.", "date": "1982", "date_type": "published", "publication": "Journal of Cyclic Nucleotide Research", "volume": "8", "number": "5", "publisher": "Raven Press", "pagerange": "309-322", "id_number": "CaltechAUTHORS:20201009-153010808", "issn": "0095-1544", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201009-153010808", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "resource_type": "article", "pub_year": "1982", "author_list": "Steer, M. L.; Braun, S.; et el." }, { "id": "https://authors.library.caltech.edu/records/74xdr-9zs55", "eprint_id": 105803, "eprint_status": "archive", "datestamp": "2023-08-19 14:35:08", "lastmod": "2023-10-20 22:27:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "A potpourri of plucked patches", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1981 Nature publishing Group.", "abstract": "HAMILL, Marty, Neher, Sakmann and\nSigworth and Horn and Patlak recently described new techniques for improved recording from single ion channels in biological membranes. They also showed how to excise the membrane patch under study, so that the experimenter can manipulate the solution bathing either the cytoplasmic or the external face. These methods, described in a News and Views article by McBurney , are leading to an explosion of new knowledge. The pace can be gauged by this review of six months' progress.", "date": "1981-12-03", "date_type": "published", "publication": "Nature", "volume": "294", "number": "5840", "publisher": "Nature Publishing Group", "pagerange": "398-399", "id_number": "CaltechAUTHORS:20201005-113706566", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113706566", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1038/294398a0", "resource_type": "article", "pub_year": "1981", "author_list": "Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/t5q9j-6ja37", "eprint_id": 105802, "eprint_status": "archive", "datestamp": "2023-08-19 13:18:26", "lastmod": "2023-10-20 22:26:58", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Nass-M-M", "name": { "family": "Nass", "given": "Menasche M." } }, { "id": "Krouse-M-E", "name": { "family": "Krouse", "given": "Mauri E." } }, { "id": "Nerbonne-J-M", "name": { "family": "Nerbonne", "given": "Jeanne M." } }, { "id": "Wassermann-N-H", "name": { "family": "Wassermann", "given": "Norbert H." } }, { "id": "Erlanger-B-F", "name": { "family": "Erlanger", "given": "Bernard F." } } ] }, "title": "Electrophysiological Experiments With Photoisomerizable Cholinergic Compounds: Review and Progress Report", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1980, NYAS.\n\nThis research was supported by the Muscular Dystrophy Association (fellowship to M.M.N. and grant\u2010in\u2010aid), by the National Institutes of Health (fellowship to J.M.N., RCDA NS\u2010272 to H.A.L., and grant no. NS\u201011756), and by the National Science Foundation (grant no. PCM\u201074\u201002140).", "abstract": "Many small molecules, both natural and synthetic, act on biological membranes by controlling ionic channels in the membrane. Our particular interest is the channel associated with the acetylcholine receptor in the postsynaptic membrane of the nicotinic synapse, where impulses are transferred from a nerve to a muscle fiber or (in some fishes) to an electroplaque. This synapse is a highly efficient electrochemical machine, specialized to function on a millisecond time scale. The immediate challenge is to understand the electrical and chemical regulation of receptor channels on this same time scale.", "date": "1980-06", "date_type": "published", "publication": "Annals of the New York Academy of Sciences", "volume": "346", "publisher": "New York Academy of Sciences", "pagerange": "475-490", "id_number": "CaltechAUTHORS:20201005-113706477", "issn": "0077-8923", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113706477", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Muscular Dystrophy Association" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "NIH", "grant_number": "NS\u2010272" }, { "agency": "NIH", "grant_number": "NS\u201011756" }, { "agency": "NSF", "grant_number": "PCM 74\u201002140" } ] }, "doi": "10.1111/j.1749-6632.1980.tb22118.x", "resource_type": "article", "pub_year": "1980", "author_list": "Lester, Henry A.; Nass, Menasche M.; et el." }, { "id": "https://authors.library.caltech.edu/records/3wd62-30q52", "eprint_id": 11551, "eprint_status": "archive", "datestamp": "2023-08-22 02:16:08", "lastmod": "2023-10-17 15:08:05", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Krouse-M-E", "name": { "family": "Krouse", "given": "Mauri E." } }, { "id": "Nass-M-M", "name": { "family": "Nass", "given": "Menasche M." } }, { "id": "Wassermann-N-H", "name": { "family": "Wassermann", "given": "Norbert H." } }, { "id": "Erlanger-B-F", "name": { "family": "Erlanger", "given": "Bernard F." } } ] }, "title": "A covalently bound photoisomerizable agonist. Comparison with reversibly bound agonists at electrophorus electroplaques", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1980 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nReceived for publication 18 June 1979. \n\nWe thank D. Williams for assisting with the animals and dissecting the cells, J.M. Nerbonne and M.M. Weinstock for discussion, and Astra Pharmaceuticals, Inc., for a gift of QX-222. This work was supported by the Muscular Dystrophy Associations of America (postdoctoral fellowship to Dr. Nass and grant-in-aid), by the National Institutes of Health (Research Career Development Award NS-272 to Dr. Lester and grant NS-11756), and by the National Science Foundation (grant PCM-74-02140).\n\nPublished - LESjgp80.pdf
", "abstract": "After disulphide bonds are reduced with dithiothreitol, trans-3-(alpha-bromomethyl)-3'-[alpha-(trimethylammonium)methyl]azobenzene (trans-QBr) alkylates a sulfhydryl group on receptors. The membrane conductance induced by this \"tethered agonist\" shares many properties with that induced by reversible agonists. Equilibrium conductance increases as the membrane potential is made more negative; the voltage sensitivity resembles that seen with 50 [mu]M carbachol. Voltage- jump relaxations follow an exponential time-course; the rate constants are about twice as large as those seen with 50 mu M carbachol and have the same voltage and temperature sensitivity. With reversible agonists, the rate of channel opening increases with the frequency of agonist-receptor collisions: with tethered trans-Qbr, this rate depends only on intramolecular events. In comparison to the conductance induced by reversible agonists, the QBr-induced conductance is at least 10-fold less sensitive to competitive blockade by tubocurarine and roughly as sensitive to \"open-channel blockade\" bu QX-222. Light-flash experiments with tethered QBr resemble those with the reversible photoisomerizable agonist, 3,3',bis-[alpha-(trimethylammonium)methyl]azobenzene (Bis-Q): the conductance is increased by cis {arrow} trans photoisomerizations and decreased by trans {arrow} cis photoisomerizations. As with Bis-Q, ligh-flash relaxations have the same rate constant as voltage-jump relaxations. Receptors with tethered trans isomer. By comparing the agonist-induced conductance with the cis/tans ratio, we conclude that each channel's activation is determined by the configuration of a single tethered QBr molecule. The QBr-induced conductance shows slow decreases (time constant, several hundred milliseconds), which can be partially reversed by flashes. The similarities suggest that the same rate-limiting step governs the opening and closing of channels for both reversible and tethered agonists. Therefore, this step is probably not the initial encounter between agonist and receptor molecules.", "date": "1980-02", "date_type": "published", "publication": "Journal of General Physiology", "volume": "75", "number": "2", "publisher": "Rockefeller University Press", "pagerange": "207-232", "id_number": "CaltechAUTHORS:LESjgp80", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LESjgp80", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "Muscular Dystrophy Associations of America" }, { "agency": "NIH", "grant_number": "NS-272" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NSF", "grant_number": "PCM-74-02140" } ] }, "doi": "10.1085/jgp.75.2.207", "pmcid": "PMC2215740", "primary_object": { "basename": "LESjgp80.pdf", "url": "https://authors.library.caltech.edu/records/3wd62-30q52/files/LESjgp80.pdf" }, "resource_type": "article", "pub_year": "1980", "author_list": "Lester, Henry A.; Krouse, Mauri E.; et el." }, { "id": "https://authors.library.caltech.edu/records/x2w6b-e7y33", "eprint_id": 105801, "eprint_status": "archive", "datestamp": "2023-08-19 12:41:55", "lastmod": "2023-10-20 22:26:49", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Armstrong-D-L", "name": { "family": "Armstrong", "given": "David L." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "The kinetics of tubocurarine action and restricted diffusion within the synaptic cleft", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1979 The Physiological Society. \n\n(Received 7 September 1978) \n\nWe are grateful to A. J. Hudspeth, S. R. Levinson, M. N. Nass, R. E. Sheridan and F. Strumwasser for their valuable discussion during these experiments, and to D. H. Jenkinson, D. D. Koblin and R. D. Purves for improving the manuscript. The work was supported by grants from the NIH (NS-11756) and the Muscular Dystrophy Association, Inc., and by an Alfred P. Sloan Research Fellowship and an NIH Research Career Development Award (NS-272) to H.A.L.", "abstract": "The kinetics of tubocurarine inhibition were studied at the post\u2010synaptic membrane of frog skeletal muscle fibres. Acetylcholine (ACh) and (+)\u2010tubocurarine were ionophoresed from twin\u2010barrel micropipettes, and the membrane potential of the muscle fibre was recorded intracellularly. Tubocurarine\u2010receptor binding was measured by decreases in the response to identical pulses of ACh. 2. The responses to both ACh and tubocurarine had brief latencies and reached their maxima rapidly. It is suggested that under these conditions the kinetics of tubocurarine action are not slowed by diffusion in the space outside the synaptic cleft. 3. After a pulse of tubocurarine, recovery from inhibition proceeds along a roughly exponential time course with a rate constant, 1/tau off approximately equal to 0.5 sec\u207b\u00b9. This rate constant does not depend on the maximal level of inhibition and varies only slightly with temperature (Q\u2081\u2080 = 1.25). 4. After a sudden maintained increase in tubocurarine release, the ACh responses decrease and eventually reach a new steady\u2010state level. Inhibition develops exponentially with time and the apparent rate constant, 1/tau on, is greater than 1/tau off. When the steady\u2010state inhibition reduces the ACh response to 1/n of its original level, the data are summarized by the relation, 1/tau on = n(1/tau off). 5. When the ACh sensitivity is reduced with cobra toxin, both 1/tau on and 1/tau off increase. Thus, the kinetics of tubocurarine inhibition depend on the density of ACh receptors in the synaptic cleft. 6. After treatment with collagenase, part of the nerve terminal is displaced and the post\u2010synaptic membrane is exposed directly to the external solution. Under these circumstances, 1/tau off increases more than tenfold. 7. Bath\u2010applied tubocurarine competitively inhibits the responses to brief ionophoretic ACh pulses with an apparent equilibrium dissociation constant, K = 0.5 microM. 8. In denervated frog muscle fibres, extrasynaptic receptors have a lower apparent affinity for tubocurarine. After a pulse of tubocurarine, inhibition decays tenfold more rapidly at these extrasynaptic sites than at the synapse. 9. It is suggested that each tubocurarine molecule binds repeatedly to several ACh receptors before escaping from the synaptic from the synaptic cleft and that the probability of this repetitive binding is enhanced because the nerve terminal presents a physical barrier to diffusion out of the cleft. Consequently, the receptor transiently buffer the concentration of tubocurarine in the cleft, and the macroscopic kinetics of inhibition are much slower than the molecular binding rates for tubocurarine.", "date": "1979-09-01", "date_type": "published", "publication": "Journal of Physiology", "volume": "294", "number": "1", "publisher": "Physiological Society", "pagerange": "365-386", "id_number": "CaltechAUTHORS:20201005-113706376", "issn": "0022-3751", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113706376", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Alfred P. Sloan Foundation" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" } ] }, "doi": "10.1113/jphysiol.1979.sp012935", "pmcid": "PMC1280562", "resource_type": "article", "pub_year": "1979", "author_list": "Armstrong, David L. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/45pm4-zm735", "eprint_id": 105800, "eprint_status": "archive", "datestamp": "2023-08-19 12:37:36", "lastmod": "2023-10-20 22:26:46", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Krouse-M-E", "name": { "family": "Krouse", "given": "Mauri E." } }, { "id": "Nass-M-M", "name": { "family": "Nass", "given": "Menasche M." } }, { "id": "Wassermann-N-H", "name": { "family": "Wassermann", "given": "Norbert H." } }, { "id": "Erlanger-B-F", "name": { "family": "Erlanger", "given": "Bernard F." } } ] }, "title": "Light-activated drug confirms a mechanism of ion channel blockade", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1979 Nature Publishing Group. \n\nReceived 05 March 1979. Accepted 06 June 1979. \n\nWe thank D. Williams for performing the dissections, M. Walsh for help with the equipment, and J. Nerbonne for discussion. Supported by the NIH (RCDA NS-272 to H.A.L. and grant NS-11756), by the Muscular Dystrophy Association (postdoctoral fellowship to M.M.N. and grant-in-aid), and by the NSF (grant PCM 74-02140).", "abstract": "A variety of mechanisms underlie the pharmacological blockade of membrane excitability. Some drugs seem to reduce the frequency at which ion channels open; a good example is the effect of curare on acetylcholine receptor channels at normal resting potentials. Another sort of mechanism may account for the action of many local anaesthetics and related drugs containing charged ammonium groups. It is postulated that such molecules block transmembrane currents as they bind to sites within open ion channels, much like a plug in a drain, with the important difference that the events occur on a millisecond time scale. This model, which we shall call 'open-channel blockade', was first applied to the effect of internal tetraethylammonium ions on K\u207a channels in squid axon and more recently to similar actions of local anaesthetics on acetylcholine receptor channels and on electrically excitable Na\u207a channels. (Curare seems to exert an additional open-channel blockade at high negative potentials.) The concept of open-channel blockade would receive direct experimental support from the demonstration that the blockade is exerted even if the blocking molecule is not bound to the channel (or indeed is not present at all) until after the channel opens. Such a demonstration is made possible by a drug that (1) blocks acetylcholine receptor channels in Electrophorus electroplaque, and (2) is created, in less than a millisecond, by a flash of light.", "date": "1979-08", "date_type": "published", "publication": "Nature", "volume": "280", "number": "5722", "publisher": "Nature Publishing Group", "pagerange": "509-510", "id_number": "CaltechAUTHORS:20201005-113706274", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113706274", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" }, { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "NSF", "grant_number": "PCM 74-02140" } ] }, "doi": "10.1038/280509a0", "resource_type": "article", "pub_year": "1979", "author_list": "Lester, Henry A.; Krouse, Mauri E.; et el." }, { "id": "https://authors.library.caltech.edu/records/axvnb-nke39", "eprint_id": 105799, "eprint_status": "archive", "datestamp": "2023-08-19 12:33:03", "lastmod": "2023-10-20 22:26:38", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Wathey-J-C", "name": { "family": "Wathey", "given": "John C." } }, { "id": "Nass-M-M", "name": { "family": "Nass", "given": "Menasche M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Numerical reconstruction of the quantal event at nicotinic synapses", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1979 The Biophysical Society. Published by Elsevier Under an Elsevier user license. \n\nReceived for publication 30 November 1978 and in revised form 10 February 1979. \n\nWe thank D. L. Armstrong, M. E. Krouse, T. L. Rosenberry, M. M. Salpeter, and R. E. Sheridan for helpful discussion. \n\nThis work was supported by the Muscular Dystrophy Association of America (postdoctoral fellowship to M.M.N. and grant-in-aid) and by the U.S. National Institutes of Health (Research Career Development Award NS-272 to H.A.L. and grant NS-11756).", "abstract": "To test our present quantitative knowledge of nicotinic transmission, we reconstruct the postsynaptic conductance change that results after a presynaptic nerve terminal liberates a quantum of acetylcholine (ACh) into the synaptic cleft. The theory assumes that ACh appears suddenly in the cleft and that is subsequent fate is determined by radial diffusion, by enzymatic hydrolysis, and by binding to receptors. Each receptor has one channel and two ACh binding sites; the channel opens when both sites are occupied and the rate-limiting step id the binding and dissociation of the second ACh molecule. The calculations reproduce the experimentally measured growth phase (200 microseconds), peak number of open channels (2,000), and exponential decay phase. The time constant of the decay phase exceeds the channel duration by approximately equal to 20%. The normal event is highly localized: at the peak, two-thirds of the open channels are within an area of 0.15 micrometer 2. This represents 75% of the available channels within this area. The model also simulates voltage and temperature dependence and effects of inactivating esterase and receptors. The calculations show that in the absence of esterase, transmitter is buffered by binding to receptors and the postsynaptic response can be potentiated.", "date": "1979-07", "date_type": "published", "publication": "Biophysical Journal", "volume": "27", "number": "1", "publisher": "Biophysical Society", "pagerange": "145-164", "id_number": "CaltechAUTHORS:20201005-113706172", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113706172", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Muscular Dystrophy Association" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1016/s0006-3495(79)85208-x", "pmcid": "PMC1328553", "resource_type": "article", "pub_year": "1979", "author_list": "Wathey, John C.; Nass, Menasche M.; et el." }, { "id": "https://authors.library.caltech.edu/records/wnfbf-1xf49", "eprint_id": 105792, "eprint_status": "archive", "datestamp": "2023-08-19 12:25:55", "lastmod": "2023-10-20 22:26:00", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Koblin-D-D", "name": { "family": "Koblin", "given": "Donald D." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Voltage-dependent and voltage-independent blockade of acetylcholine receptors by local anesthetics in Electrophorus electroplaques", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1979 by Academic Press. \n\n(Received June 19, 1978) (Accepted November 21, 1978) \n\nThis work was supported by National Institutes of Health Grant NS-11756; by a Grant-in-Aid from the Muscular Dystrophy Association, Inc.; by a Muscular Dystrophy Postdoctoral Fellowship to D.D.K.; and by an Alfred P. Sloan Fellowship and N.I.H. Career Development Award NS272 to H.A.L.", "abstract": "This study employs voltage-clamp techniques to survey the action of several local anesthetics on the nicotinic acetylcholine receptor of Electrophorus electroplaques. These drugs partially block the steady-state activation of receptors during bath application of agonist. Certain anesthetics alter the kinetics of voltage-jump relaxations during such application and alter the waveform of neurally-evoked postsynaptic currents. Procaine, tetracaine, and QX-222 act similarly at concentrations of 25-50 \u00b5M. When the agonist is carbamylcholine, acetylcholine, or suberyldicholine, the steady-state blockade depends on voltage, with a greater fractional inhibition of agonist-induced currents at more negative membrane potential. Alterations of voltage-jump relaxations are most noticeable with suberyldicholine. Under some conditions, at least two exponential relaxation components are seen; one component is much faster and another much slower than the single component seen in the absence of procaine. In contrast, the percentage blockade by dibucaine and chlorpromazine is about the same at all membrane potentials, and these compounds have little influence on suberyldicholine relaxation kinetics. The uncharged local anesthetic, benzocaine, at a concentration ten times that of the other local anesthetics employed (250 \u00b5M), only slightly decreases suberyldicholine-induced currents and has little effect on the relaxation rates. Thus, voltage affects the relative potencies of local anesthetics in blocking agonist-induced currents. At low negative membrane potentials, the potency of the anesthetics parallels their lipid solubiity. At high negative membrane potentials this correlation disappears. The results suggest a dual mode of action for the local anesthetics: an indirect interaction with the lipids surrounding the receptor, and a direct, voltage-dependent interaction with the receptor-channel complex.", "date": "1979-05-01", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "15", "number": "3", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "559-580", "id_number": "CaltechAUTHORS:20201005-113630529", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113630529", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Alfred P. Sloan Foundation" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" } ] }, "resource_type": "article", "pub_year": "1979", "author_list": "Koblin, Donald D. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/aykm1-qbb17", "eprint_id": 11552, "eprint_status": "archive", "datestamp": "2023-08-22 01:53:20", "lastmod": "2023-10-17 15:08:07", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Analysis of sodium and potassium redistribution during sustained permeability increases at the innervated face of Electrophorus electroplaques", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1978 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nReceived for publication 24 March 1978. \n\nI thank D. Armstrong, P. Ascher, M. Brodwick, J-P. Changeux, J.B. Cohen, D.D. Koblin, M.M. Nass, J-L. Popot, R.E. Sheridan, F. Strumwasser, and D. Van Essen for helpful discussion. E.X. Albuquerque supplied a sample of batrachotoxin. \n\nThis research was supported by the National Institutes of Health (Research Grant NS-11756 and Research Career Development Award NS-272) and by a grant from the Muscular Dystrophy Association.\n\nPublished - LESjgp78.pdf
", "abstract": "Cholinergic agonists cause an increase in the membrane permeability of Na and K at the innervated face of Electrophorus electroplaques. Therefore, sustained exposure to agonist reduces Na and K concentration gradients. There gradients are monitored with voltage-clamp sequences and pharmacological treatments that selectively measure the Nernst potentials for individual ions. EK is normally near--90 mV but moves toward zero during bath application of agonist. Depolarizations by bath- applied agonist measure primarily this shift of EK, not short- circuiting of EK by the agonist-induced conductance. After a rapid jump of agonist concentration, there is a fast (millisecond) depolarization due to the conductance increase, followed by a much slower additional \"creep\" due to the shift in EK. Sodium replaces the lost intracellular potassium: ENa, normally very positive, also moves toward zero. The shifts in EK and ENa are normally reversible but become permanent after blockade of the Na-K pump. In the presence of agonist, the shifts can be driven further by passing current of the appropriate polarity. Similar ion redistribution occurs with other drugs, such as batrachotoxin and nystatin, which induce prolonged increases in Na permeability. The redistributions cause little net change in the reversal potential of the neurally evoked postsynaptic current.", "date": "1978-12", "date_type": "published", "publication": "Journal of General Physiology", "volume": "72", "number": "6", "publisher": "Rockefeller University Press", "pagerange": "857-862", "id_number": "CaltechAUTHORS:LESjgp78", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LESjgp78", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-1175" }, { "agency": "NIH", "grant_number": "NS-272" }, { "agency": "Muscular Dystrophy Association" } ] }, "doi": "10.1085/jgp.72.6.847", "pmcid": "PMC2228490", "primary_object": { "basename": "LESjgp78.pdf", "url": "https://authors.library.caltech.edu/records/aykm1-qbb17/files/LESjgp78.pdf" }, "resource_type": "article", "pub_year": "1978", "author_list": "Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/8wj8h-s5150", "eprint_id": 105798, "eprint_status": "archive", "datestamp": "2023-08-19 11:40:35", "lastmod": "2023-10-20 22:26:35", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Nass-M-M", "name": { "family": "Nass", "given": "Menasche M." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Krouse-M-E", "name": { "family": "Krouse", "given": "Mauri E." } } ] }, "title": "Response of acetylcholine receptors to photoisomerizations of bound agonist molecules", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1978 The Biophysical Society. Published by Elsevier Under an Elsevier user license. \n\nReceived for publication 1 December 1977. \n\nWe thank D. Williams for help with the animals, M. Walsh for help with the optics and electronics, H. W. Chang for furnishing the Bis-Q, and D. Armstrong, R. K. Clayton, B. F. Erlanger, R. E. Sheridan, and N. H. Wassermann for helpful discussion. \n\nThis research was supported by a postdoctoral fellowship from the Muscular Dystrophy Association of America to M.M.N., by an Alfred P. Sloan Research Fellowship and National Institutes of Health Career Development Award (NS-272) to H.A.L., by an N.I.H. predoctoral training grant to M.E.K., and by research grants from the Muscular Dystrophy Association and the N.I.H. (NS-11756).", "abstract": "In these experiments, agonist-induced conductance is measured while a sudden perturbation is produced at the agonist-receptor binding site. A voltage-clamped Electrophorus electroplaque is exposed to trans-Bis-Q, a potent agonist. Some channels are open; these receptors have bound agonist molecules. A light flash isomerizes 3(-35)% of the trans-Bis-Q molecules to their cis form, a far poorer agonist. This causes a rapid decrease of membrane conductance (phase 1), followed by a slower increase (phase 2). Phase 1 has the amplitude and wavelength dependence expected if the channel closes within 100 mus after a single bound trans-Bis-Q is isomerized, and if the photochemistry of bound Bis-Q resembles that in solution. Therefore, the receptor channel responds rapidly, and with a hundred-fold greater closing rate, after this change in the structure of a bound ligand. Phase 2 (the conductance increase) seems to represent the relaxation back toward equilibrium after phase 1, because (a) phase 2 has the same time constant (1(-5) ms) as a voltage- or concentration-jump relaxation under identical conditions; and (b) phase 2 is smaller if the flash has led to a net decrease in (trans-Bis-Q). Still slower signals follow: phase 3, a decrease of conductance (time constant 5(-10 ms); and phase 4, an equal and opposite increase (several seconds). Phase 3 is abolished by curare and does not depend on the history of the membrane voltage. We consider several mechanisms for phases 3 and 4.", "date": "1978-10", "date_type": "published", "publication": "Biophysical Journal", "volume": "24", "number": "1", "publisher": "Biophysical Society", "pagerange": "135-160", "id_number": "CaltechAUTHORS:20201005-113706084", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113706084", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "Muscular Dystrophy Association of America" }, { "agency": "Alfred P. Sloan Foundation" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" }, { "agency": "NIH Predoctoral Fellowship" }, { "agency": "NIH", "grant_number": "NS-11756" } ] }, "doi": "10.1016/s0006-3495(78)85352-1", "pmcid": "PMC1473870", "resource_type": "article", "pub_year": "1978", "author_list": "Nass, Menasche M.; Lester, Henry A.; et el." }, { "id": "https://authors.library.caltech.edu/records/986tv-wxr27", "eprint_id": 105797, "eprint_status": "archive", "datestamp": "2023-08-19 11:17:54", "lastmod": "2023-10-20 22:26:29", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Koblin-D-D", "name": { "family": "Koblin", "given": "Donald D." } }, { "id": "Sheridan-R-E", "name": { "family": "Sheridan", "given": "Robert E." } } ] }, "title": "Role of voltage-sensitive receptors in nicotinic transmission", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1978 The Biophysical Society. Published by Elsevier Under an Elsevier user license. \n\nReceived for publication 23 June 1977 and in revised form 10 November 1977. \n\nWe thank D. Williams for assistance with the animals and dissections, and P. Hartig, J. Heuser, M. Salpeter, and R. Zucker for helpful discussion. \n\nThis work was supported by National Institutes of Health grant NS-11756; by a Grant-in-Aid from the Muscular Dystrophy Association, Inc.; by a Muscular Dystrophy Fellowship to D.D.K.; by an Alfred P. Sloan Fellowship and NIH Career Development Award NS272 to H.A.L.; and by a Spencer Foundation Fellowship to R.E.S.", "abstract": "This paper compares the conductance induced by bath-applied acetyl-choline (ACh) and by the same transmitter released from nerve terminals at Electrophorus electroplaques. For the former case, dose-response relations are characterized by the maximal agonist-induced conductance, r\u03b3 (130 mmho/cm\u00b2), and by the concentration which induces half this conductance; this concentration is termed K_(app) and equals 50 micron at -85 mV. For the latter case, neurally evoked postsynaptic currents (PSCs) are characterized by the peak conductance during strongly facilitated release, g_(PSC), and by the rate constant for decay, \u03b1. Since g_(PSC) roughly equals r\u03b3, it is concluded that the PSC activates nearly all available receptor channels. These and other data agree with recent estimates that during the growth phase of the quantal response, (a) the ACh concentration is at least several hundred micromolar; and (b) most nearby channels are activated. However both \u03b1 and K_(app) increase during depolarization, at a rate of about e-fold per 86 mV. These observations on voltage sensitivity suggest that a suprathreshold synaptic event is rapidly terminated because the action potential abruptly releases ACh molecules from receptors.", "date": "1978-03", "date_type": "published", "publication": "Biophysical Journal", "volume": "21", "number": "3", "publisher": "Biophysical Society", "pagerange": "181-194", "id_number": "CaltechAUTHORS:20201005-113705979", "issn": "0006-3495", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113705979", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Alfred P. Sloan Foundation" }, { "agency": "NIH Postdoctoral Fellowship", "grant_number": "NS-272" }, { "agency": "Spencer Foundation" } ] }, "doi": "10.1016/s0006-3495(78)85518-0", "pmcid": "PMC1473691", "resource_type": "article", "pub_year": "1978", "author_list": "Lester, Henry A.; Koblin, Donald D.; et el." }, { "id": "https://authors.library.caltech.edu/records/jkb4n-9pg07", "eprint_id": 28863, "eprint_status": "archive", "datestamp": "2023-08-19 10:48:12", "lastmod": "2023-10-24 18:12:03", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Sheridan-R-E", "name": { "family": "Sheridan", "given": "Robert E." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Rates and equilibria at the acetylcholine receptor of electrophorus electroplaques. A study of neurally evoked postsynaptic currents and of voltage-jump relaxations", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1977 Rockefeller University Press.\n\nReceived for publication 12 July 1976.\n\nWe thank D. Williams for assistance with the animals and dissections, W. L. Crowder and P. Goodeve for help with the computer, F. Sigworth for building the voltage clamp, J. Heesemann for a gift of suberyldicholine, E. Neher for a discussion of edge effects, and D. D. Koblin for\nparticipating in some of the experiments. This work was supported by National Institutes of Health grant NS-11756; by a grant-in-aid from the Muscular Dystrophy Association, Inc.; by an Alfred P. Sloan fellowship to H. A. Lester; and by a National Science Foundation predoctoral fellowship to R. E. Sheridan.\n\nPublished - SHEjgp97.pdf
", "abstract": "Kinetic measurements are employed to reconstruct the steady-state activation of acetylcholine [Ach] receptor channels in electrophorus electroplaques. Neurally evoked postsynaptic currents (PSCs) decay exponentially; at 15\u00b0C the rate constant, \u03b1, equals 1.2 ms^(-1) at 0 mV and decreases e-fold for every 86 mV as the membrane voltage is made more negative. Voltage-jump relaxations have been measured with bath-applied ACh, decamethonium, carbachol, or suberylcholine. We interpret the reciprocal relaxation time 1/\u03c4 as the sum of the rate constant \u03b1 for channel closing and a first-order rate constant for channel opening. Where measureable, the opening rate increases linearly with [agonist] and does not vary with voltage. The voltage sensitivity of small steady-state conductances (e- fold for 86 mV) equals that of the closing rate \u03b1, confirming that the opening rate has little or no additional voltage sensitivity. Exposure to \u03b1-bungarotoxin irreversibly decreases the agonist-induced conductance but does not affect the relaxation kinetics. Tubocurarine reversibly reduces both the conductance and the opening rate. In the simultaneous presence of two agonist species, voltage-jump relaxations have at least two exponential components. The data are fit by a model in which (a) the channel opens as the receptor binds the second in a sequence of two agonist molecules, with a forward rate constant to 10^(7) to 2x10^(8) M^(-1)s^(-1); and (b) the channel then closes as either agonist molecule dissociates, with a voltage-dependent rate constant of 10^(2) to 3x10^(3)s^(-1).", "date": "1977-08-01", "date_type": "published", "publication": "Journal of General Physiology", "volume": "70", "number": "2", "publisher": "Rockefeller University Press", "pagerange": "187-219", "id_number": "CaltechAUTHORS:20120119-120416762", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20120119-120416762", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Alfred P. Sloan Foundation" }, { "agency": "NSF Graduate Research Fellowship" } ] }, "doi": "10.1085/jgp.70.2.187", "pmcid": "PMC2228462", "primary_object": { "basename": "SHEjgp97.pdf", "url": "https://authors.library.caltech.edu/records/jkb4n-9pg07/files/SHEjgp97.pdf" }, "resource_type": "article", "pub_year": "1977", "author_list": "Sheridan, Robert E. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/t76st-3gw20", "eprint_id": 105796, "eprint_status": "archive", "datestamp": "2023-08-19 10:33:14", "lastmod": "2023-10-20 22:26:21", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Chang-Hai Won", "name": { "family": "Chang", "given": "Hai Won" } } ] }, "title": "Response of acetylcholine receptors to rapid photochemically produced increases in agonist concentration", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 Nature Publishing Group 1977. \n\nReceived 01 November 1976. Accepted 25 January 1977. \n\nWe thank D. Williams for assistance with the animals, W. L. Crowder for help with electronics, D. D. Koblin for participating in some of the experiments, M. A. Raftery for the use of a spectrophotometer, E. Neumann for helpful discussion, and the Chadwick-Helmuth Corporation (Monrovia, California) for the loan of a flash tube and power supply. This work was supported by the NIH, by the Muscular Dystrophy Association, and by an Alfred P. Sloan Fellowship to H.A.L.", "abstract": "SYNAPTIC transmission to muscle fibres and electroplaques takes place on a millisecond timescale. To study the molecular events leading to activation of the cholinergic receptor channel, we have developed a method to produce similarly rapid, spatially uniform steps of agonist concentration near intact postsynaptic membranes. Electrophorus electroplaques in a voltage-clamp apparatus were exposed to a solution containing a photochromic compound. Initially the compound was in a predominantly cis form, which had little effect on the membrane. During a brief light flash, some of the compound was isomerised to the trans isomer, which is a cholinergic agonist. As a result the membrane conductance increased along a timecourse which reflects the rate processes underlying activation of the cholinergic receptor channel.", "date": "1977-03-24", "date_type": "published", "publication": "Nature", "volume": "266", "number": "5600", "publisher": "Nature Publishing Group", "pagerange": "373-374", "id_number": "CaltechAUTHORS:20201005-113705890", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113705890", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH" }, { "agency": "Muscular Dystrophy Association" }, { "agency": "Alfred P. Sloan Foundation" } ] }, "doi": "10.1038/266373a0", "resource_type": "article", "pub_year": "1977", "author_list": "Lester, Henry A. and Chang, Hai Won" }, { "id": "https://authors.library.caltech.edu/records/5y1s9-93342", "eprint_id": 105795, "eprint_status": "archive", "datestamp": "2023-08-19 10:28:21", "lastmod": "2023-10-20 22:26:18", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "The response to acetylcholine", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1977 Scientific American.", "abstract": "When a nerve makes a muscle cell contract, it gives the cell a tiny squirt of acetylcholine. Receptors on the cell respond by opening so that ions can travel through the cell membrane.", "date": "1977-02", "date_type": "published", "publication": "Scientific American", "volume": "236", "number": "2", "publisher": "Scientific American", "pagerange": "106-118", "id_number": "CaltechAUTHORS:20201005-113705797", "issn": "0036-8733", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113705797", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "doi": "10.1038/scientificamerican0277-106", "resource_type": "article", "pub_year": "1977", "author_list": "Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/gx6ts-ne038", "eprint_id": 7740, "eprint_status": "archive", "datestamp": "2023-08-22 00:53:42", "lastmod": "2023-10-16 21:04:37", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Sheridan-R-E", "name": { "family": "Sheridan", "given": "Robert E." } }, { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Relaxation measurements on the acetylcholine receptor", "ispublished": "pub", "full_text_status": "public", "keywords": "synaptic transmission; voltage clamp; ion channels; Electrophorus electroplaque", "note": "\u00a9 1975 by the National Academy of Sciences. \n\nCommunicated by James Olds, June 25, 1975. \n\nWe thank F. Sigworth for constructing the voltage-clamp circuit, D. Williams for technical assistance, and N.J.M. Birdsall, J.-P. Changeux, T. Jovin, W. Newsome, M.A. Raftery, and G. Zweig for helpful discussion. This work was supported by NIH Grant NS-11756, by an NSF Predoctoral Fellowship (to R.E.S.), and by an Alfred P. Sloan Fellowship (to H.A.L.).\n\nPublished - SHEpnas75.pdf
", "abstract": "In Electrophorus electroplaques, the agonist-induced postsynaptic conductance depends on membrane potential. During steady exposure to agonists, after a voltage step the conductance relaxes on a millisecond time scale, exponentially approaching a new equilibrium value. The relaxation rate constant k is an instantaneous function of voltage, insensitive to the past or present conductance. \n\nTwo components sum to form k. A concentration-sensitive component increases linearly with agonist concentration and decreases during desensitization or exposure to curare. Thus this component reflects the average frequency at which acetylcholine receptors are opening. \n\nThe voltage-sensitive component, obtained by extrapolating k to zero agonist concentration, increases at more positive potentials. For acetylcholine, the voltage-sensitive component equals the rate constant for the exponential decay of postsynaptic currents; it thus seems to be the closing rate for active receptors. The voltage-sensitive component has the relative amplitudes acetylcholine < carbamoylcholine < decamethonium, and for each agonist equals the closing rate determined from \"noise\" measurements at neuromuscular junctions. \n\nThe kinetic data explain several aspects of the steady-state conductance induced by agonists, but shed no light on apparent cooperative effects.", "date": "1975-09", "date_type": "published", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "volume": "72", "number": "9", "publisher": "National Academy of Sciences", "pagerange": "3496-3500", "id_number": "CaltechAUTHORS:SHEpnas75", "issn": "0027-8424", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:SHEpnas75", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "NSF Predoctoral Fellowship" }, { "agency": "Alfred P. Sloan Foundation" } ] }, "doi": "10.1073/pnas.72.9.3496", "pmcid": "PMC433021", "primary_object": { "basename": "SHEpnas75.pdf", "url": "https://authors.library.caltech.edu/records/gx6ts-ne038/files/SHEpnas75.pdf" }, "resource_type": "article", "pub_year": "1975", "author_list": "Sheridan, Robert E. and Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/nw2k8-70188", "eprint_id": 11553, "eprint_status": "archive", "datestamp": "2023-08-22 00:49:57", "lastmod": "2023-10-17 15:08:10", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" }, { "id": "Changeux-J-P", "name": { "family": "Changeux", "given": "Jean-Pierre" } }, { "id": "Sheridan-R-E", "name": { "family": "Sheridan", "given": "Robert E." } } ] }, "title": "Conductance increases produced by bath application of cholinergic agonists to Electrophorus electroplaques", "ispublished": "pub", "full_text_status": "public", "note": "\u00a9 1975 by The Rockefeller University Press. RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode. \n\nReceived for publication 15 November 1974. \n\nWe thank Mmes. S. Meugon and H. Nuret for expert assistance and P. Ascher, E.A. Barnard, H. Buc, J. B. Cohen, F.A. Dodge, A. Marry, R. Olsen, J.-L. Popot, and M. Weber for fruitful discussions. F. Sigworth built an improved voltage-clamp circuit for use in the instantaneous experiments. The investigations were supported by the U.S. National Institutes of Health (postdoctoral fellowship and grant NS-11756 to H.A.L.) and National Science Foundation (predoctoral Fellowship to R.E.S.), the Centre National de la Recherche Scientifique, the D\u00e9l\u00e9gation G\u00e9n\u00e9rale a la Recherche Scientifique et Technique, the Fondation pour la Recherche M\u00e9dicale Fran\u00e7aise, the Coll\u00e8ge de France, and the Commissariat \u00e0 l'Energie Atomique.\n\nPublished - LESjgp75.pdf
", "abstract": "When solutions containing agonists are applied to the innervated face of an Electrophorus electroplaque, the membrane's conductance increases. The agonist-induced conductance is increased at more negative membrane potentials. The \"instantaneous\" current-voltage curve for agonist-induced currents is linear and shows a reversal potential near zero mV; chord conductances, calculated on the basis of this reversal potential, change epsilon-fold for every 62-mV change in potential when the conductance is small. Conductance depends non- linearly on small agonist concentrations; at all potentials, the dose-response curve has a Hill coefficient of 1.45 for decamethonium (Deca) and 1.90 for carbamylcholine (Carb). With agonist concentrations greater than 10^(-4) M Carb or 10^(-5) M Deca, the conductance rises to a peak 0.5-1.5 min after introduction of agonist, then declines with time; this effect resembles the \"desensitization\" reported for myoneural junctions. Elapid alpha-toxin, tubocurarine, and desensitization reduce the conductance without changing the effects of potential; the apparent dissociation constant for tubocurarine is 2 X 10^(-7) M. By contrast, procaine effects a greater fractional inhibition of the conductance at high negative potentials.", "date": "1975-06", "date_type": "published", "publication": "Journal of General Physiology", "volume": "65", "number": "6", "publisher": "Rockefeller University Press", "pagerange": "797-816", "id_number": "CaltechAUTHORS:LESjgp75", "issn": "0022-1295", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:LESjgp75", "rights": "RUP grants the public the non-exclusive right to copy, distribute, or display the Work under a Creative Commons Attribution\u2013Noncommercial\u2013Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/ and http://creativecommons.org/licenses/by-nc-sa/3.0/legalcode.", "funders": { "items": [ { "agency": "NIH", "grant_number": "NS-11756" }, { "agency": "Centre National de la Recherche Scientifique (CNRS)" }, { "agency": "D\u00e9l\u00e9gation G\u00e9n\u00e9rale a la Recherche Scientifique et Technique" }, { "agency": "Fondation pour la Recherche M\u00e9dicale Fran\u00e7aise" }, { "agency": "Coll\u00e8ge de France" }, { "agency": "Commissariat \u00e0 l'Energie Atomique (CEA)" } ] }, "doi": "10.1085/jgp.65.6.797", "pmcid": "PMC2214893", "primary_object": { "basename": "LESjgp75.pdf", "url": "https://authors.library.caltech.edu/records/nw2k8-70188/files/LESjgp75.pdf" }, "resource_type": "article", "pub_year": "1975", "author_list": "Lester, Henry A.; Changeux, Jean-Pierre; et el." }, { "id": "https://authors.library.caltech.edu/records/kk471-tst60", "eprint_id": 105790, "eprint_status": "archive", "datestamp": "2023-08-19 08:07:39", "lastmod": "2023-10-20 22:25:52", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Blockade of acetylcholine receptors by cobra toxin: electrophysiological studies", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1972 by Academic Press. \n\n(Received March 14, 1972) \n\nI am indebted to F. A. Dodge, Jr., and to F. Ratliff, my predoctoral advisors, for their unfailing guidance. I also thank C. M. Connelly, H. K. Hartline, A. Mauro, and D. Mauzerall for advice. H. P. Rang helped with preparation of the manuscript. \n\nThese studies were supported by Grant EY 188 from the National Institutes of Health, Grant GB 6540 from the National Science Foundation, and a Predoctoral Fellowship from the National Institutes of Health, United States Public Health Service.", "abstract": "Microelectrode techniques were used to investigate the manner in which a pure polypeptide toxin from cobra venom blocks transmission at the frog myoneural junction. At concentrations of 16-114 nM, the toxin causes an irreversible exponential decline in the amplitude of the end plate potential as a result of a decrease in the sensitivity of the postsynaptic receptors for acetylcholine. Other processes, such as spontaneous and impulse-evoked acetylcholine release, acetylcholinesterase activity, and passive electrical properties of the muscle fiber membrane, remain unaffected. The rate constant for inactivation of receptors increases linearly with toxin concentration. The constant of proportionality probably describes the binding of toxin to the receptor and equals 1.5 x 10\u2075 M\u207b\u00b9 sec\u207b\u00b9. Certain complications in these experiments arise because diffusion barriers limit the access of toxin molecules to the receptors.", "date": "1972-11-01", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "8", "number": "6", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "623-631", "id_number": "CaltechAUTHORS:20201005-113628628", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113628628", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "EY 188" }, { "agency": "NSF", "grant_number": "GB 6540" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "resource_type": "article", "pub_year": "1972", "author_list": "Lester, H. A." }, { "id": "https://authors.library.caltech.edu/records/342dx-ecw66", "eprint_id": 105791, "eprint_status": "archive", "datestamp": "2023-08-19 08:07:47", "lastmod": "2023-10-20 22:25:57", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "H. A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Vulnerability of desensitized or curare-treated acetylcholine receptors to irreversible blockade by cobra toxin", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9, 1972, by Academic Press. \n\n(Received March 14, 1972) \n\nI am grateful to F. A. Dodge, Jr., and F. Ratliff, my predoctoral advisors. D. Colquhoun, D. Eaker, H. K. Hartline, B. W. Knight, A. Mauro, and D. Mauzerall also contributed insights. H. P. Rang advised on the preparation of the manuscript. \n\nThese studies were supported by Grant EY 188 from the National Institutes of Health, Grant GB 6544) from the National Science Foundation, and a Graduate Fellowship from the National Institutes of Health, United States Public Health Service.", "abstract": "Inactivation of frog myoneural acetylcholine receptors by cobra toxin (24-120 nM) is studied in the presence of reversible antagonists (d-tubocurarine, dihydro-\u03b2-erythroidine) and agonists (carbachol, nicotine). During agonist treatments most receptors are densensitized rather than activated. Neither agonists nor antagonists reverse the blockade produced by the toxin. \n\nAt 26 \u00b5M d-tubocurarine, receptors are partially protected from the toxin. At 5 \u00b5M d-tubocurarine, a concentration which blocks 97% of the receptors, or lower concentrations, curare-blocked and free receptors are equally vulnerable to the toxin. \n\nReceptors are protected against the toxin at 7-140 \u00b5M carbachol, concentrations which desensitize 60-99.6% of the receptors. During combined treatment with agonist and toxin, end plate potential amplitudes decline in 300-600 sec to an irreversible \"plateau\" rather than exponentially to zero. The plateau is well described, as a very slow decline in the number of available receptors, by the following scheme. (A) The receptor population undergoes three simultaneous processes with first-order rate constants in the range 10-4 to 10-2 sec-1: reversible desensitization by agonists, recovery from desensitization, and irreversible inactivation by toxin. (B) Desensitized receptors are essentially invulnerable to the toxin. \n\nThe agonist data are consistent with the model for desensitization proposed by Katz and Thesleff [J. Physiol. (London) 138, 63 (1957)].", "date": "1972-11-01", "date_type": "published", "publication": "Molecular Pharmacology", "volume": "8", "number": "6", "publisher": "American Society for Pharmacology and Experimental Therapeutics", "pagerange": "632-644", "id_number": "CaltechAUTHORS:20201005-113629507", "issn": "0026-895X", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113629507", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "EY 188" }, { "agency": "NSF", "grant_number": "GB 6544" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "resource_type": "article", "pub_year": "1972", "author_list": "Lester, H. A." }, { "id": "https://authors.library.caltech.edu/records/bfwgc-9b482", "eprint_id": 105794, "eprint_status": "archive", "datestamp": "2023-08-19 07:12:18", "lastmod": "2023-10-20 22:26:12", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Postsynaptic Action of Cobra Toxin at the Myoneural Junction", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1970 Nature Publishing Group. \n\nReceived April 2, 1970. \n\nthank Mr D. Eaker for toxin and Mr F. A. Dodge, Mr. A. Mauro, and Mr F. Ratliff for advice. This work was supported by grants from tho National Institutes of Neurological Diseases and Blindness and the National Science Foundation and by a predoctoral fellowship from the National Institutes of Health.", "abstract": "Recent investigations of peptide fractions from cobra venoms suggest that the most potent acute toxicity arises from a curare-like block of the action of acetylcholine at the skeletal myoneural junction. I wish to report the effects on myoneural transmission of toxin T\u2083 from Naja naja siamensis\u2014(Kaouthia) (Thailand cobra) venom, a pure peptide of known amino-acid composition.", "date": "1970-08-15", "date_type": "published", "publication": "Nature", "volume": "227", "number": "5259", "publisher": "Nature Publishing Group", "pagerange": "727-728", "id_number": "CaltechAUTHORS:20201005-113705712", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113705712", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NSF" }, { "agency": "NIH Predoctoral Fellowship" } ] }, "doi": "10.1038/227727a0", "resource_type": "article", "pub_year": "1970", "author_list": "Lester, Henry A." }, { "id": "https://authors.library.caltech.edu/records/1cagg-q6w43", "eprint_id": 105793, "eprint_status": "archive", "datestamp": "2023-08-19 07:11:41", "lastmod": "2023-10-20 22:26:06", "type": "article", "metadata_visibility": "show", "creators": { "items": [ { "id": "Lester-H-A", "name": { "family": "Lester", "given": "Henry A." }, "orcid": "0000-0002-5470-5255" } ] }, "title": "Transmitter Release by Presynaptic Impulses in the Squid Stellate Ganglion", "ispublished": "pub", "full_text_status": "restricted", "note": "\u00a9 1970 Nature Publishing Group. \n\nReceived January 27; revised March 2, 1970. \n\nI thank F. A. Dodge, J. P. Hervey and F. Ratliff for advice and discussion and H. K. Hartline for support under NIH and NSF grants; NIH provided predoctoral fellowship support.", "abstract": "In spite of phylogenetic differences, the vertebrate (frog) myoneural junction and the giant synapse in the squid stellate ganglion have similar characteristics of transmitter release. In this article I report quantitative data on this similarity, on the roles of Ca\u00b2\u207a and Mg\u00b2\u207a, and the dependence on temperature.", "date": "1970-08-01", "date_type": "published", "publication": "Nature", "volume": "227", "number": "5257", "publisher": "Nature Publishing Group", "pagerange": "493-496", "id_number": "CaltechAUTHORS:20201005-113705571", "issn": "0028-0836", "official_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201005-113705571", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH Predoctoral Fellowship" }, { "agency": "NSF" } ] }, "doi": "10.1038/227493a0", "resource_type": "article", "pub_year": "1970", "author_list": "Lester, Henry A." } ]