Artificial intelligence (AI) is rapidly transforming stem cell and developmental biology, offering new strategies to analyze, interpret and optimize complex, dynamic systems such as organoids and stem cell-derived embryo models. In this Perspective, we chart the integration of AI into image-based analysis of stem cell systems, highlighting how deep learning, convolutional neural networks and emerging foundation models enable automated classification, segmentation and phenotyping at increasing scale and precision. We showcase applications in phenotyping, drug screening and mechanistic discovery, including real-time fate prediction and the identification of hidden morphological signatures linked to differentiation and disease. Practical challenges, including limited annotated data, model interpretability and live imaging constraints, are examined alongside future opportunities, such as multimodal integration, real-time experimental steering and protocol optimization. Altogether, we argue that AI is not merely an analytical tool, but a discovery engine that enhances reproducibility, accelerates insight and brings us closer to a mechanistic understanding of self-organization in complex stem cell-derived systems.
", "doi": "10.1038/s41592-026-03202-x", "issn": "1548-7091", "publisher": "Nature Publishing Group", "publication": "Nature Methods", "publication_date": "2026-08-14", "volume": "23", "pages": "1710\u20131723" }, { "id": "authors:qdw5t-n4d40", "collection": "authors", "collection_id": "qdw5t-n4d40", "cite_using_url": "https://authors.library.caltech.edu/records/qdw5t-n4d40", "type": "article", "title": "Genetic recording and in situ readout of single-cell signaling memory", "author": [ { "family_name": "Hao", "given_name": "Kai", "orcid": "0000-0002-8458-3404" }, { "family_name": "Liu", "given_name": "Yunzheng", "orcid": "0009-0003-0559-7440", "clpid": "Liu-Yunzheng" }, { "family_name": "Barrett", "given_name": "Mykel", "orcid": "0000-0003-4620-4116" }, { "family_name": "Samadi", "given_name": "Zainalabedin" }, { "family_name": "Zarezadeh", "given_name": "Amirhossein", "orcid": "0009-0002-1010-691X" }, { "family_name": "McGrath", "given_name": "Yuka" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Askary", "given_name": "Amjad", "orcid": "0000-0002-2913-8498" } ], "abstract": "Intensity and duration of biological signals encode a few pathways to direct diverse cellular behaviors, yet quantifying these features in single cells remains difficult. To address this challenge, we developed INSCRIBE, which uses a CRISPR base editor to mutate genomic targets at rates proportional to signaling activity. Edits are recovered at the endpoint through a new ratiometric readout strategy from images of two fluorescence channels. We engineered human cells to record WNT and BMP activity. Following defined exogenous stimulations, INSCRIBE accurately recovered signal intensity in dose\u2013response experiments and exposure duration in time-course experiments. Applying INSCRIBE revealed a persistent memory in the BMP pathway, where progeny of high-responding cells remained more sensitive to subsequent BMP stimulation for up to 3 weeks. Together, our results establish a scalable platform for genetic recording and in situ readout of signaling activity in single cells, advancing quantitative analysis of cell\u2013cell communication during development and disease.", "doi": "10.1038/s41589-026-02168-3", "issn": "1552-4450", "publisher": "Nature Publishing Group", "publication": "Nature Chemical Biology", "publication_date": "2026-03-11" }, { "id": "authors:136v6-w4h72", "collection": "authors", "collection_id": "136v6-w4h72", "cite_using_url": "https://authors.library.caltech.edu/records/136v6-w4h72", "type": "article", "title": "Using deep learning to predict the sex of human embryos", "author": [ { "family_name": "Liang", "given_name": "Mingshu" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Lamba", "given_name": "Adiyant" }, { "family_name": "Gokyer", "given_name": "Dilan" }, { "family_name": "King", "given_name": "Catherine" }, { "family_name": "Meglicki", "given_name": "Maciej" }, { "family_name": "Pardi\u00f1as", "given_name": "Mar\u00eda Luisa" }, { "family_name": "de Los Santos", "given_name": "Mar\u00eda Jos\u00e9" }, { "family_name": "Yang", "given_name": "Changhuei" } ], "abstract": "The existence of sex differences in human pre-implantation development remains an open question, with previous attempts based on human observations yielding inconclusive results. In this study, we combined manual annotation and deep learning analysis of a dataset comprising 515 time-lapse embryo movies to investigate whether birth sex influences early developmental dynamics. While manual assessment did not identify any developmental timing parameters that reliably distinguish male from female embryos, a deep learning model trained and tested on these videos achieves a statistically significant sex prediction accuracy of 61%. Importantly, our analyses identified the period after the eight-cell stage as critical for accurate prediction, indicating that subtle sex-related differences may begin to emerge around day 3 of human embryogenesis. Studying sex differences at this early stage may enhance our understanding of why some embryos fail to develop and why sex ratios can be skewed in the context of in vitro fertilization. More broadly, our findings open the possibility for an early, non-invasive detection tool that could assist in identifying and addressing sex-related embryonic developmental abnormalities.
", "doi": "10.1098/rsob.250419", "issn": "2046-2441", "publisher": "The Royal Society", "publication": "Open Biology", "publication_date": "2026-03-01", "series_number": "3", "volume": "16", "issue": "3", "pages": "250419" }, { "id": "authors:pmvqp-gwx39", "collection": "authors", "collection_id": "pmvqp-gwx39", "cite_using_url": "https://authors.library.caltech.edu/records/pmvqp-gwx39", "type": "article", "title": "John Gurdon 1933\u20132025", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "John Gurdon taught us that the beginning of life is never truly lost — only waiting to be reawakened. Long before his ideas were accepted and celebrated, he showed that a cell remembers more than it appears to know: that within a differentiated nucleus lies the latent capacity to begin anew. This insight, pursued with patience and extraordinary experimental care, would ultimately reshape the field of developmental biology. John’s approach to science arose from a lifelong curiosity about how life organizes itself, renews itself and surprises us. And, to those who knew John well, his science was inseparable from his character — gentle, persistent, quietly radical.
", "doi": "10.1038/s41587-026-03015-5", "issn": "1087-0156", "publisher": "Nature Publishing Group", "publication": "Nature Biotechnology", "publication_date": "2026-02", "volume": "44", "pages": "181\u2013182" }, { "id": "authors:v27bb-hhe02", "collection": "authors", "collection_id": "v27bb-hhe02", "cite_using_url": "https://authors.library.caltech.edu/records/v27bb-hhe02", "type": "article", "title": "Efficient stem cell-derived mouse embryo models for environmental studies", "author": [ { "family_name": "Jorgensen", "given_name": "Victoria", "orcid": "0000-0002-4205-6198", "clpid": "Jorgensen-Victoria-Lynn" }, { "family_name": "Bao", "given_name": "Min", "orcid": "0009-0002-4247-3511", "clpid": "Bao-Min" }, { "family_name": "Junyent", "given_name": "Sergi", "orcid": "0000-0003-2405-5885", "clpid": "Junyent-Sergi" }, { "family_name": "H\u00e4felfinger", "given_name": "Christoph M.", "orcid": "0009-0000-2800-9144", "clpid": "H\u00e4felfinger-Christoph-M" }, { "family_name": "Amaya", "given_name": "Laura", "orcid": "0000-0002-8742-9111", "clpid": "Amaya-Laura" }, { "family_name": "Liao", "given_name": "Zhaodi", "orcid": "0000-0002-5387-732X", "clpid": "Liao-Zhaodi" }, { "family_name": "Williams", "given_name": "Brian A.", "orcid": "0000-0003-3253-611X", "clpid": "Williams-Brian-A" }, { "family_name": "Chen", "given_name": "Dong-Yuan", "orcid": "0000-0003-2179-2847", "clpid": "Chen-Dong-Yuan" }, { "family_name": "Wu", "given_name": "Amanda", "orcid": "0000-0001-9546-1472" }, { "family_name": "Thomson", "given_name": "Matt", "orcid": "0000-0003-1021-1234", "clpid": "Thomson-M-W" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Blastoids are stem cell-derived structures that mimic natural blastocysts by incorporating all three lineages: trophectoderm, epiblast, and primitive endoderm. However, current methods often yield incomplete structures that fail to cavitate or to form a proper primitive endoderm. To overcome these limitations, we develop a modular approach by aggregating three murine stem cell types: embryonic stem cells (ESCs), ESCs with inducible GATA4 expression (iG4-ESCs), and trophoblast stem cells (TSCs). This method yields cavitated blastocyst-like structures-termed iG4-blastoids-with approximately 80% efficiency. Single-cell RNA sequencing confirms their close resemblance to mature mouse blastocysts. Notably, culturing iG4-blastoids without FGF4 enhances specification of the invasive mural trophectoderm, and approximately 12% of structures undergo post-implantation-like morphogenesis in vitro. Using this model, we show that caffeine, alcohol, nicotine, and amino acid variations affect iG4-blastoids and natural embryos similarly, underscoring their utility as a robust model for investigating the impact of diverse environmental factors on embryogenesis.", "doi": "10.1016/j.devcel.2025.08.004", "issn": "1534-5807", "publisher": "Elsevier", "publication": "Developmental Cell", "publication_date": "2026-01-14", "series_number": "1", "volume": "61", "issue": "1", "pages": "193-207.e6" }, { "id": "authors:sexh8-37h83", "collection": "authors", "collection_id": "sexh8-37h83", "cite_using_url": "https://authors.library.caltech.edu/records/sexh8-37h83", "type": "article", "title": "Asynchronous mouse embryo polarization leads to heterogeneity in cell fate specification", "author": [ { "family_name": "Lamba", "given_name": "Adiyant", "orcid": "0000-0001-5389-1730" }, { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840" }, { "family_name": "Meglicki", "given_name": "Maciej", "orcid": "0009-0000-9224-3795" }, { "family_name": "Czukiewska", "given_name": "Sylwia", "orcid": "0000-0001-9151-9008" }, { "family_name": "Balasubramaniam", "given_name": "Lakshmi", "orcid": "0000-0001-6881-8261" }, { "family_name": "Hadas", "given_name": "Ron", "orcid": "0000-0001-9301-6607", "clpid": "Hadas-Ron" }, { "family_name": "Weishaupt", "given_name": "Nina" }, { "family_name": "Patel", "given_name": "Ekta M", "clpid": "Patel-Ekta-M" }, { "family_name": "Kavanagh", "given_name": "Yu Hua", "orcid": "0009-0004-0882-6998" }, { "family_name": "Wang", "given_name": "Ran" }, { "family_name": "Jing", "given_name": "Naihe", "orcid": "0000-0003-1509-6378" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The first lineage allocation in mouse and human embryos separates the inner cell mass (ICM) from the outer trophectoderm (TE). This symmetry-breaking event is executed through polarization of cells at the 8 cell stage and subsequent asymmetric divisions, generating polar (TE) and apolar (ICM) cells. Here, we show that mouse embryo polarization is unexpectedly asynchronous. Cells polarizing at the early and late 8 cell stage have distinct molecular and morphological properties that direct their following lineage specification, with early polarizing cells being biased towards producing the TE lineage. More recent studies have also implicated heterogeneities between cells prior to the 8 cell stage in the first lineage allocation: cells exhibiting reduced methyltransferase CARM1 activity at the 4 cell stage are predisposed towards the TE fate. Here, we demonstrate that reduced CARM1 activity and upregulation of its substrate BAF155 promote early polarization and TE specification. These findings provide a link between asymmetries at the 4 cell stage and polarization at the 8 cell stage, mechanisms of the first lineage allocation that had been considered separate.", "doi": "10.7554/elife.101140", "issn": "2050-084X", "publisher": "eLife Sciences Publications, Ltd", "publication": "eLife", "publication_date": "2025-12-08", "volume": "13", "pages": "RP101140" }, { "id": "authors:mb1xy-59651", "collection": "authors", "collection_id": "mb1xy-59651", "cite_using_url": "https://authors.library.caltech.edu/records/mb1xy-59651", "type": "article", "title": "Fertilization triggers early proteomic symmetry breaking in mammalian embryos", "author": [ { "family_name": "Iwamoto-Stohl", "given_name": "Lisa K.", "orcid": "0000-0003-0776-054X", "clpid": "Iwamoto-Stohl-Lisa-K" }, { "family_name": "Petelski", "given_name": "Aleksandra A." }, { "family_name": "Quan", "given_name": "Baiyi", "orcid": "0000-0001-6313-4274", "clpid": "Quan-Baiyi" }, { "family_name": "Meglicki", "given_name": "Maciej" }, { "family_name": "Fu", "given_name": "Audrey" }, { "family_name": "Nakagawa", "given_name": "Shoma", "orcid": "0000-0002-7766-613X", "clpid": "Nakagawa-Shoma" }, { "family_name": "McMahon", "given_name": "Breanna", "orcid": "0009-0001-3618-9664", "clpid": "McMahon-Breanna-M" }, { "family_name": "Wang", "given_name": "Ting-Yu", "orcid": "0000-0002-9014-6825", "clpid": "Wang-Ting-Yu" }, { "family_name": "Khan", "given_name": "Saad" }, { "family_name": "Specht", "given_name": "Harrison" }, { "family_name": "Huffman", "given_name": "Gray" }, { "family_name": "Derks", "given_name": "Jason" }, { "family_name": "Junyent", "given_name": "Sergi", "orcid": "0000-0003-2405-5885", "clpid": "Junyent-Sergi" }, { "family_name": "Weatherbee", "given_name": "Bailey A.T." }, { "family_name": "Weberling", "given_name": "Antonia" }, { "family_name": "Gantner", "given_name": "Carlos W." }, { "family_name": "Mandelbaum", "given_name": "Rachel S." }, { "family_name": "Paulson", "given_name": "Richard J." }, { "family_name": "Lam", "given_name": "Lisa" }, { "family_name": "Chou", "given_name": "Tsui-Fen", "orcid": "0000-0003-2410-2186", "clpid": "Chou-Tsui-Fen" }, { "family_name": "Slavov", "given_name": "Nikolai" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "While non-mammalian embryos often rely on spatial pre-patterning, mammalian development has long been thought to begin with equivalent blastomeres. However, emerging evidence challenges this. Here, using multiplexed and label-free single-cell proteomics, we identify over 300 asymmetrically abundant proteins—many involved in protein degradation and transport—dividing mouse 2-cell-stage blastomeres into two distinct clusters, which we term alpha and beta. These proteomic asymmetries are detectable as early as the zygote stage, intensify by the 4-cell stage, and correlate with the sperm entry site, implicating fertilization as a symmetry-breaking event. Splitting 2-cell-stage embryos into halves reveals that beta blastomeres possess greater developmental potential than alpha blastomeres. Similar clustering and protein enrichment patterns found in human 2-cell embryos suggest this early asymmetry might be conserved. These findings uncover a previously unrecognized proteomic pre-patterning triggered by fertilization in mammalian embryos, with important implications for understanding totipotency and early lineage bias.
", "doi": "10.1016/j.cell.2025.11.006", "issn": "0092-8674", "publisher": "Cell Press", "publication": "Cell", "publication_date": "2025-12-03" }, { "id": "authors:tfxmd-bwd37", "collection": "authors", "collection_id": "tfxmd-bwd37", "cite_using_url": "https://authors.library.caltech.edu/records/tfxmd-bwd37", "type": "article", "title": "Role of CEBPa in trophectoderm competence installment", "author": [ { "family_name": "Wei", "given_name": "Xiao", "orcid": "0000-0001-5150-7483" }, { "family_name": "Salvador-Martinez", "given_name": "Irepan", "orcid": "0000-0002-3188-9784" }, { "family_name": "Meglicki", "given_name": "Maciej", "orcid": "0009-0000-9224-3795" }, { "family_name": "Plana-Carmona", "given_name": "Marcos", "orcid": "0000-0002-1976-7506" }, { "family_name": "Klonizakis", "given_name": "Antonios", "orcid": "0000-0001-9850-6971" }, { "family_name": "Pernaute", "given_name": "Barbara", "orcid": "0000-0002-8706-1126" }, { "family_name": "Irimia", "given_name": "Manuel", "orcid": "0000-0002-2179-2567" }, { "family_name": "Stik", "given_name": "Gregoire", "orcid": "0000-0002-1404-1992" }, { "family_name": "Popovic", "given_name": "Mina", "orcid": "0000-0001-9657-0611" }, { "family_name": "Garcia", "given_name": "Guillem Torcal", "orcid": "0000-0002-0843-7296" }, { "family_name": "Heyn", "given_name": "Holger", "orcid": "0000-0002-3276-1889" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Graf", "given_name": "Thomas", "orcid": "0000-0003-2774-4117" } ], "abstract": "During mouse embryogenesis, totipotency is gradually lost, and, at the 16-cell stage, blastomeres begin to bifurcate into trophectoderm (future placenta) and inner cell mass (future fetus). Although this process is well studied, when and how blastomeres acquire the competence for lineage specification remains unclear. Here, we describe that CEBPa becomes up-regulated at the transition from the two- to the four-cell stage by NR5A2 and is also selectively expressed in the trophectoderm at the blastocyst stage. Its knockout decreases the proportion of trophectoderm cells and delays the morula to blastocyst transition. Conversely, CEBPa overexpression in mouse embryonic stem cells, used as a proxy, drives their differentiation into trophectoderm-like cells, enabling the identification of CEBPa-regulated trophectoderm-specific enhancers. A subset of these enhancers, associated with key trophectoderm-related transcription factor genes, is primed or activated in four- and eight-cell embryos. Together, our data suggest that CEBPa plays a role in the installment of trophectoderm competence before the first lineage bifurcation and in trophectoderm specification.", "doi": "10.1126/sciadv.ady1693", "pmcid": "PMC12662221", "issn": "2375-2548", "publisher": "American Association for the Advancement of Science", "publication": "Science Advances", "publication_date": "2025-11-28", "series_number": "48", "volume": "11", "issue": "48", "pages": "eady1693" }, { "id": "authors:rkhm5-hpr97", "collection": "authors", "collection_id": "rkhm5-hpr97", "cite_using_url": "https://authors.library.caltech.edu/records/rkhm5-hpr97", "type": "article", "title": "HIF1A-mediated pathways promote euploid cell survival in chromosomally mosaic embryos", "author": [ { "family_name": "Sanchez-Vasquez", "given_name": "Estefania", "orcid": "0000-0002-6585-8548", "clpid": "Sanchez-Vasquez-Estefania" }, { "family_name": "Bronner", "given_name": "Marianne E", "orcid": "0000-0003-4274-1862", "clpid": "Bronner-M-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Human fertility is suboptimal in part by error-prone divisions during early cleavage stages, which frequently result in chromosomal aneuploidy. Most human pre-implantation embryos are mosaics of euploid and aneuploid cells, yet those with a low proportion of aneuploid cells can develop to term at rates similar to fully euploid embryos. How embryos manage aneuploidy during early development remains poorly understood – yet this knowledge is crucial for improving fertility outcomes and reducing developmental defects. To investigate these mechanisms, we established a new mouse model of chromosome mosaicism to trace the fate of aneuploid cells during pre-implantation development. We previously used the Mps1 inhibitor reversine to induce aneuploidy. Here, we demonstrate that the more specific Mps1 inhibitor AZ3146 similarly disrupts chromosome segregation but supports higher developmental potential than reversine. AZ3146-treated embryos transiently upregulate hypoxia-inducible factor-1A (HIF1A) without triggering Trp53 activation. Given that pre-implantation embryos develop in a hypoxic environment in vivo, we further explored the role of oxygen tension. Hypoxia exposure in vitro reduced DNA damage in response to Mps1 inhibition and increased the proportion of euploid cells in mosaic epiblast. Conversely, HIF1A inhibition decreased the proportion of aneuploid cells. Together, these findings uncover a role for hypoxia signaling in modulating the response to chromosomal errors and suggest new strategies to improve the developmental potential of mosaic human embryos.
", "doi": "10.7554/elife.101912", "issn": "2050-084X", "publisher": "eLife Sciences Publications, Ltd", "publication": "eLife", "publication_date": "2025-10-14", "volume": "13", "pages": "RP101912" }, { "id": "authors:2ma17-86y37", "collection": "authors", "collection_id": "2ma17-86y37", "cite_using_url": "https://authors.library.caltech.edu/records/2ma17-86y37", "type": "article", "title": "Spindle-localized F-actin regulates polar MTOC organization and the fidelity of meiotic spindle formation", "author": [ { "family_name": "Soto-Moreno", "given_name": "Edgar J." }, { "family_name": "Ali", "given_name": "Nourhan N." }, { "family_name": "K\u00fcllmer", "given_name": "Florian", "orcid": "0000-0002-0671-1546" }, { "family_name": "Nasufovic", "given_name": "Veselin", "orcid": "0000-0001-6812-637X" }, { "family_name": "Frolikova", "given_name": "Michaela", "orcid": "0000-0002-9834-8405" }, { "family_name": "Tepla", "given_name": "Olga" }, { "family_name": "Masata", "given_name": "Jaromir" }, { "family_name": "Trauner", "given_name": "Dirk", "orcid": "0000-0002-6782-6056" }, { "family_name": "Patterson", "given_name": "Amanda A." }, { "family_name": "Arndt", "given_name": "Hans-Dieter", "orcid": "0000-0002-0792-1422" }, { "family_name": "Komrskova", "given_name": "Katerina", "orcid": "0000-0002-6837-2148" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Balboula", "given_name": "Ahmed Z.", "orcid": "0000-0001-9663-6262" } ], "abstract": "Mammalian oocytes are notoriously prone to chromosome segregation errors leading to aneuploidy. The spindle provides the machinery for accurate chromosome segregation during cell division. Mammalian oocytes lack centrioles and, therefore, mouse meiotic spindle relies on the organization of numerous acentriolar microtubule organizing centers into two poles (polar microtubule organizing centers, pMTOCs). The traditional view is that, in mammalian oocytes, microtubules are the sole cytoskeletal component responsible for regulating pMTOC organization and spindle assembly. We identify a previously unrecognized F-actin pool that surrounds pMTOCs, forming F-actin cage-like structure. We demonstrate that F-actin localization on the spindle depends on unconventional myosins X and VIIb. Selective disruption of spindle-localized F-actin, using myosin X/VIIb knockdown oocytes or photoswitchable Optojasp-1, perturbs pMTOC organization, leading to unfocused spindle poles and chromosome missegregation. Here, we unveil an important function of spindle-localized\u00a0F-actin in regulating pMTOC organization, a critical process for ensuring the fidelity of meiotic spindle formation and proper chromosome segregation.", "doi": "10.1038/s41467-025-63586-w", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2025-09-19", "series_number": "1", "volume": "16", "issue": "1", "pages": "8323" }, { "id": "authors:wp1g5-a0413", "collection": "authors", "collection_id": "wp1g5-a0413", "cite_using_url": "https://authors.library.caltech.edu/records/wp1g5-a0413", "type": "article", "title": "Basement membrane perforations guide anterior\u2013posterior axis formation", "author": [ { "family_name": "Chen", "given_name": "Dong-Yuan", "orcid": "0000-0003-2179-2847" }, { "family_name": "Claussen", "given_name": "Nikolas H.", "orcid": "0000-0002-9020-6437" }, { "family_name": "Titus", "given_name": "Shiny" }, { "family_name": "Hu", "given_name": "Wenqi", "clpid": "Hu-Wenqi" }, { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278" }, { "family_name": "Mandelbaum", "given_name": "Rachel S.", "orcid": "0000-0001-8611-1462" }, { "family_name": "Scott Jr", "given_name": "Richard T." }, { "family_name": "Seli", "given_name": "Emre" }, { "family_name": "Streichan", "given_name": "Sebastian J.", "orcid": "0000-0002-6105-9087" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Establishment of the anterior-posterior (AP) axis is a critical symmetry-breaking event in mammalian development. In mice, this process involves the directed migration of the distal visceral endoderm (DVE). Here, we use targeted perturbations to demonstrate that asymmetric perforations in the basement membrane guide DVE migration. During implantation, matrix metalloproteinases in extra-embryonic tissues create uneven basement membrane perforations, establishing directional cues for cohesive DVE migration. Using light-sheet microscopy and tissue cartography, we show that migrating DVE deforms surrounding tissues. Physical modeling and live imaging of DVE protrusions indicate that basement membrane perforations orchestrate active force generation within the DVE. Extending these findings to human embryos and stem cell-derived models, we identify basement membranes with enriched perforations near the anterior hypoblast in embryos, suggesting a conserved mechanism for AP axis specification. These findings reveal an unrecognized role of basement membrane remodeling and mechanical heterogeneity in guiding directional tissue migration during mammalian development.", "doi": "10.1038/s41467-025-61441-6", "pmcid": "PMC12284222", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2025-07-22", "series_number": "1", "volume": "16", "issue": "1", "pages": "6763" }, { "id": "authors:rryjh-r3641", "collection": "authors", "collection_id": "rryjh-r3641", "cite_using_url": "https://authors.library.caltech.edu/records/rryjh-r3641", "type": "article", "title": "Atlas of amnion development during the first trimester of human pregnancy", "author": [ { "family_name": "Hu", "given_name": "Wenqi", "orcid": "0000-0002-7934-9429", "clpid": "Hu-Wenqi" }, { "family_name": "Sancho-Serra", "given_name": "Carmen" }, { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786" }, { "family_name": "Szafranska", "given_name": "Hanna M.", "clpid": "Szafranska-Hanna-M" }, { "family_name": "Solanky", "given_name": "Nita", "orcid": "0000-0001-8304-5543" }, { "family_name": "Metcalfe", "given_name": "Kate" }, { "family_name": "Vento-Tormo", "given_name": "Roser" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The amnion is a critical extra-embryonic structure that supports foetal development, yet its ontogeny remains poorly defined. Here, using single-cell transcriptomics, we identified major cell types and subtypes in the human amnion across the first trimester of pregnancy, broadly categorized into epithelial, mesenchymal and macrophage lineages. We uncovered epithelial–mesenchymal and epithelial–immune transitions, highlighting dynamic remodelling during early pregnancy. Our results further revealed key intercellular communication pathways, including BMP4 signalling from mesenchymal to epithelial cells and TGF-β signalling from macrophages to mesenchymal cells, suggesting coordinated interactions that drive amnion morphogenesis. In addition, integrative comparisons across humans, non-human primates and in vitro stem cell-based models reveal that stem cell-based models recapitulate various stages of amnion development, emphasizing the need for careful selection of model systems to accurately recapitulate in vivo amnion formation. Collectively, our findings provide a detailed view of amnion cellular composition and interactions, advancing our understanding of its developmental role and regenerative potential.
", "doi": "10.1038/s41556-025-01696-9", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2025-07-14", "volume": "27", "pages": "1175\u20131185" }, { "id": "authors:fs39e-xeb27", "collection": "authors", "collection_id": "fs39e-xeb27", "cite_using_url": "https://authors.library.caltech.edu/records/fs39e-xeb27", "type": "article", "title": "AI-based approach to dissect the variability of mouse stem cell-derived embryo models", "author": [ { "family_name": "Caldarelli", "given_name": "Paolo", "orcid": "0000-0003-4455-7778", "clpid": "Caldarelli-Paolo" }, { "family_name": "Deininger", "given_name": "Luca", "orcid": "0009-0005-5125-6422" }, { "family_name": "Zhao", "given_name": "Shi", "clpid": "Zhao-Shi" }, { "family_name": "Panda", "given_name": "Pallavi", "orcid": "0000-0001-7572-6728", "clpid": "Panda-Pallavi" }, { "family_name": "Yang", "given_name": "Changhuei", "orcid": "0000-0001-8791-0354", "clpid": "Yang-Changhuei" }, { "family_name": "Mikut", "given_name": "Ralf", "orcid": "0000-0001-9100-5496" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Recent advances in stem cell-derived embryo models have transformed developmental biology, offering insights into embryogenesis without the constraints of natural embryos. However, variability in their development challenges research standardization. To address this, we use deep learning to enhance the reproducibility of selecting stem cell-derived embryo models. Through live imaging and AI-based models, we classify 900 mouse post-implantation stem cell-derived embryo-like structures (ETiX-embryos) into normal and abnormal categories. Our best-performing model achieves 88% accuracy at 90 h post-cell seeding and 65% accuracy at the initial cell-seeding stage, forecasting developmental trajectories. Our analysis reveals that normally developed ETiX-embryos have higher cell counts and distinct morphological features such as larger size and more compact shape. Perturbation experiments increasing initial cell numbers further supported this finding by improving normal development outcomes. This study demonstrates deep learning’s utility in improving embryo model selection and reveals critical features of ETiX-embryo self-organization, advancing consistency in this evolving field.
", "doi": "10.1038/s41467-025-56908-5", "pmcid": "PMC11839995", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2025-02-19", "series_number": "1", "volume": "16", "issue": "1", "pages": "1772" }, { "id": "authors:9hamt-naq41", "collection": "authors", "collection_id": "9hamt-naq41", "cite_using_url": "https://authors.library.caltech.edu/records/9hamt-naq41", "type": "article", "title": "Primitive to visceral endoderm maturation is essential for mouse epiblast survival beyond implantation", "author": [ { "family_name": "Weberling", "given_name": "Antonia", "orcid": "0000-0001-8282-5695" }, { "family_name": "Siriwardena", "given_name": "Dylan", "orcid": "0000-0002-9172-7947" }, { "family_name": "Penfold", "given_name": "Christopher", "orcid": "0000-0001-5823-4705" }, { "family_name": "Christodoulou", "given_name": "Neophytos", "orcid": "0000-0003-4338-5847" }, { "family_name": "Boroviak", "given_name": "Thorsten E.", "orcid": "0000-0001-8703-8949" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The implantation of the mouse blastocyst initiates a complex sequence of tissue remodeling and cell differentiation events required for morphogenesis, during which the extraembryonic primitive endoderm transitions into the visceral endoderm. Through single-cell RNA sequencing of embryos at embryonic day 5.0, shortly after implantation, we reveal that this transition is driven by dynamic signaling activities, notably the upregulation of BMP signaling and a transient increase in Sox7 expression. Embryos deficient in Hepatocyte nuclear factor-1-beta (Hnf1b-/-), a gene critical for visceral endoderm differentiation, showed an interaction between visceral endoderm and epiblast, crucial for epiblast survival. Single-cell RNA profiling of Hnf1b-/- visceral endoderm shows developmental delays and severe dysregulation in several nutrient transport pathways. Impaired glucose uptake in Hnf1b-/- embryos suggests that the activation of nutrient transport mechanisms during the primitive-to-visceral endoderm transition may be vital for post-implantation epiblast development. These findings offer new insights into the molecular regulation of early mammalian development.", "doi": "10.1016/j.isci.2024.111671", "pmcid": "PMC11761342", "issn": "2589-0042", "publisher": "Cell Press", "publication": "iScience", "publication_date": "2025-01-17", "series_number": "1", "volume": "28", "issue": "1", "pages": "111671" }, { "id": "authors:ncph6-nke14", "collection": "authors", "collection_id": "ncph6-nke14", "cite_using_url": "https://authors.library.caltech.edu/records/ncph6-nke14", "type": "article", "title": "Assembly of a stem cell-derived human postimplantation embryo model", "author": [ { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786" }, { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278" }, { "family_name": "Wang", "given_name": "Yuntao", "orcid": "0009-0001-9239-5892" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The embryonic and extraembryonic tissue interactions underlying human embryogenesis at implantation stages are not currently understood. We have generated a pluripotent stem cell-derived model that mimics aspects of peri-implantation development, allowing tractable experimentation otherwise impossible in the human embryo. Activation of the extraembryonic lineage-specific transcription factors GATA6 and SOX17 (hypoblast factors) or GATA3 and TFAP2C (encoding AP2\u03b3; trophoblast factors) in human embryonic stem (ES) cells drive conversion to extraembryonic-like cells. When combined with wild-type ES cells, self-organized embryo-like structures form in the absence of exogenous factors, termed human inducible embryoids (hiEmbryoids). The epiblast-like domain of hiEmbryoids polarizes and differentiates in response to extraembryonic-secreted extracellular matrix and morphogen cues. Extraembryonic mesenchyme, amnion and primordial germ cells are specified in hiEmbryoids in a stepwise fashion. After establishing stable inducible ES lines and converting ES cells to RSeT culture media, the protocol takes 7\u201310 d to generate hiEmbryoids. Generation of hiEmbryoids can be performed by researchers with basic expertise in stem cell culture.", "doi": "10.1038/s41596-024-01042-7", "issn": "1754-2189", "publisher": "Nature Publishing Group", "publication": "Nature Protocols", "publication_date": "2025-01", "series_number": "1", "volume": "20", "issue": "1", "pages": "67-91" }, { "id": "authors:wgy1x-2tf47", "collection": "authors", "collection_id": "wgy1x-2tf47", "cite_using_url": "https://authors.library.caltech.edu/records/wgy1x-2tf47", "type": "article", "title": "Temporal BMP4 effects on mouse embryonic and extraembryonic development", "author": [ { "family_name": "Hadas", "given_name": "Ron" }, { "family_name": "Rubinstein", "given_name": "Hernan" }, { "family_name": "Mittnenzweig", "given_name": "Markus" }, { "family_name": "Mayshar", "given_name": "Yoav" }, { "family_name": "Ben-Yair", "given_name": "Raz" }, { "family_name": "Cheng", "given_name": "Saifeng" }, { "family_name": "Aguilera-Castrejon", "given_name": "Alejandro", "orcid": "0000-0002-1339-7778" }, { "family_name": "Reines", "given_name": "Netta" }, { "family_name": "Orenbuch", "given_name": "Ayelet-Hashahar" }, { "family_name": "Lifshitz", "given_name": "Aviezer", "orcid": "0000-0002-8458-9507" }, { "family_name": "Chen", "given_name": "Dong-Yuan", "orcid": "0000-0003-2179-2847", "clpid": "Chen-Dong-Yuan" }, { "family_name": "Elowitz", "given_name": "Michael B.", "orcid": "0000-0002-1221-0967", "clpid": "Elowitz-M-B" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Hanna", "given_name": "Jacob H.", "orcid": "0000-0003-2042-9974" }, { "family_name": "Tanay", "given_name": "Amos", "orcid": "0000-0001-9419-3824" }, { "family_name": "Stelzer", "given_name": "Yonatan", "orcid": "0000-0001-9207-1479" } ], "abstract": "The developing placenta, which in mice originates through the extraembryonic ectoderm (ExE), is essential for mammalian embryonic development. Yet unbiased characterization of the differentiation dynamics of the ExE and its interactions with the embryo proper remains incomplete. Here we develop a temporal single-cell model of mouse gastrulation that maps continuous and parallel differentiation in embryonic and extraembryonic lineages. This is matched with a three-way perturbation approach to target signalling from the embryo proper, the ExE alone, or both. We show that ExE specification involves early spatial and transcriptional bifurcation of uncommitted ectoplacental cone cells and chorion progenitors. Early BMP4 signalling from chorion progenitors is required for proper differentiation of uncommitted ectoplacental cone cells and later for their specification towards trophoblast giant cells. We also find biphasic regulation by BMP4 in the embryo. The early ExE-originating BMP4 signal is necessary for proper mesoendoderm bifurcation and for allantois and primordial germ cell specification. However, commencing at embryonic day 7.5, embryo-derived BMP4 restricts the primordial germ cell pool size by favouring differentiation of their extraembryonic mesoderm precursors towards an allantois fate. ExE and embryonic tissues are therefore entangled in time, space and signalling axes, highlighting the importance of their integrated understanding and modelling in vivo and in vitro.
", "doi": "10.1038/s41586-024-07937-5", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2024-10-17", "volume": "634", "pages": "652\u2013661" }, { "id": "authors:1pjnf-anz02", "collection": "authors", "collection_id": "1pjnf-anz02", "cite_using_url": "https://authors.library.caltech.edu/records/1pjnf-anz02", "type": "article", "title": "Embryo models need consistent ethical oversight", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Hyun", "given_name": "Insoo", "orcid": "0000-0001-6605-7112", "clpid": "Hyun-Insoo" } ], "abstract": "Studies of stem-cell-derived models of human embryos can uncover obscure developmental events that cause pregnancies to fail. Studying ‘non-integrated’ models, which lack extra-embryonic lineages required to make an embryo biologically complete, can seem to pose fewer ethical challenges than do integrated models, which contain embryonic and extra-embryonic lineages (Nature 622, 469–470; 2023).
", "doi": "10.1038/d41586-024-01728-8", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2024-06-13", "series_number": "8016", "volume": "630", "issue": "8016", "pages": "305" }, { "id": "authors:x3gb1-p2s80", "collection": "authors", "collection_id": "x3gb1-p2s80", "cite_using_url": "https://authors.library.caltech.edu/records/x3gb1-p2s80", "type": "article", "title": "Tead4 and Tfap2c generate bipotency and a bistable switch in totipotent embryos to promote robust lineage diversification", "author": [ { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Meglicki", "given_name": "Maciej", "orcid": "0009-0000-9224-3795", "clpid": "Meglicki-Maciej" }, { "family_name": "Lamba", "given_name": "Adiyant", "orcid": "0000-0001-5389-1730", "clpid": "Lamba-Adiyant" }, { "family_name": "Wang", "given_name": "Peizhe", "orcid": "0000-0003-4298-2471", "clpid": "Wang-Peizhe" }, { "family_name": "Royer", "given_name": "Christophe", "orcid": "0000-0002-9882-7676", "clpid": "Royer-Christophe" }, { "family_name": "Turner", "given_name": "Karen", "clpid": "Turner-Karen" }, { "family_name": "Jauhar", "given_name": "Muhammad Abdullah", "orcid": "0009-0003-2916-5693", "clpid": "Jauhar-Muhammad--Abdullah" }, { "family_name": "Jones", "given_name": "Celine", "clpid": "Jones-Celine" }, { "family_name": "Child", "given_name": "Tim", "orcid": "0000-0001-6668-0529", "clpid": "Child-Tim" }, { "family_name": "Coward", "given_name": "Kevin", "orcid": "0000-0003-3577-4041", "clpid": "Coward-Kevin" }, { "family_name": "Na", "given_name": "Jie", "orcid": "0000-0003-1820-0548", "clpid": "Na-Jie" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The mouse and human embryo gradually loses totipotency before diversifying into the inner cell mass (ICM, future organism) and trophectoderm (TE, future placenta). The transcription factors TFAP2C and TEAD4 with activated RHOA accelerate embryo polarization. Here we show that these factors also accelerate the loss of totipotency. TFAP2C and TEAD4 paradoxically promote and inhibit Hippo signaling before lineage diversification: they drive expression of multiple Hippo regulators while also promoting apical domain formation, which inactivates Hippo. Each factor activates TE specifiers in bipotent cells, while TFAP2C also activates specifiers of the ICM fate. Asymmetric segregation of the apical domain reconciles the opposing regulation of Hippo signaling into Hippo OFF and the TE fate, or Hippo ON and the ICM fate. We propose that the bistable switch established by TFAP2C and TEAD4 is exploited to trigger robust lineage diversification in the developing embryo.
\nRetrospective lineage reconstruction of humans predicts that dramatic clonal imbalances in the body can be traced to the 2-cell stage embryo. However, whether and how such clonal asymmetries arise in the embryo is unclear. Here, we performed prospective lineage tracing of human embryos using live imaging, non-invasive cell labeling, and computational predictions to determine the contribution of each 2-cell stage blastomere to the epiblast (body), hypoblast (yolk sac), and trophectoderm (placenta). We show that the majority of epiblast cells originate from only one blastomere of the 2-cell stage embryo. We observe that only one to three cells become internalized at the 8-to-16-cell stage transition. Moreover, these internalized cells are more frequently derived from the first cell to divide at the 2-cell stage. We propose that cell division dynamics and a cell internalization bottleneck in the early embryo establish asymmetry in the clonal composition of the future human body.
\nThe blueprint of the mammalian body plan is laid out during gastrulation, when a trilaminar embryo is formed. This process entails a burst of proliferation, the ingression of embryonic epiblast cells at the primitive streak, and their priming toward primitive streak fates. How these different events are coordinated remains unknown. Here, we developed and characterized a 3D culture of self-renewing mouse embryonic cells that captures the main transcriptional and architectural features of the early gastrulating mouse epiblast. Using this system in combination with microfabrication and in vivo experiments, we found that proliferation-induced crowding triggers delamination of cells that express high levels of the apical polarity protein aPKC. Upon delamination, cells become more sensitive to Wnt signaling and upregulate the expression of primitive streak markers such as Brachyury. This mechanistic coupling between ingression and differentiation ensures that the right cell types become specified at the right place during embryonic development.
\nDevelopment requires coordinated interactions between the epiblast, which generates the embryo proper; the trophectoderm, which generates the placenta; and the hypoblast, which forms both the anterior signalling centre and the yolk sac. These interactions remain poorly understood in human embryogenesis because mechanistic studies have only recently become possible. Here we examine signalling interactions post-implantation using human embryos and stem cell models of the epiblast and hypoblast. We find anterior hypoblast specification is NODAL dependent, as in the mouse. However, while BMP inhibits anterior signalling centre specification in the mouse, it is essential for its maintenance in human. We also find contrasting requirements for BMP in the naive pre-implantation epiblast of mouse and human embryos. Finally, we show that NOTCH signalling is important for human epiblast survival. Our findings of conserved and species-specific factors that drive these early stages of embryonic development highlight the strengths of comparative species studies.
\nStem cell-based mammalian embryo models facilitate the discovery of developmental mechanisms because they are more amenable to genetic and epigenetic perturbations than natural embryos. Here, we highlight exciting recent advances that have yielded a plethora of models of embryonic development. Imperfections in these models highlight gaps in our current understanding and outline future research directions, ushering in an exciting new era for embryology.
", "doi": "10.1016/j.gde.2023.102134", "issn": "0959-437X", "publisher": "Elsevier", "publication": "Current Opinion in Genetics & Development", "publication_date": "2024-02", "volume": "84", "pages": "102134" }, { "id": "authors:yxn76-3fk50", "collection": "authors", "collection_id": "yxn76-3fk50", "cite_using_url": "https://authors.library.caltech.edu/records/yxn76-3fk50", "type": "article", "title": "The evolution of embryo models", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The creation of multiple stem-cell-derived models of mammalian embryogenesis is opening many new doors to study human development and brings a need for scientists to demonstrate responsible dialog over the associated ethical issues.
", "doi": "10.1038/s41592-023-02077-6", "issn": "1548-7091", "publisher": "Nature Publishing Group", "publication": "Nature Methods", "publication_date": "2023-12-06" }, { "id": "authors:xq10b-71b55", "collection": "authors", "collection_id": "xq10b-71b55", "cite_using_url": "https://authors.library.caltech.edu/records/xq10b-71b55", "type": "article", "title": "Assembly of complete mouse embryo models from embryonic and induced stem cell types in vitro", "author": [ { "family_name": "Lau", "given_name": "Kasey Y. C.", "clpid": "Lau-Kasey-Y-C" }, { "family_name": "Amadei", "given_name": "Gianluca", "orcid": "0000-0001-5405-968X", "clpid": "Amadei-Gianluca" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The interaction between embryonic and extraembryonic tissues is critical in natural mouse embryogenesis. Here, to enable such interaction in vitro, we describe a protocol to assemble a complete mouse embryo model using mouse embryonic stem cells and induced embryonic stem cells to express Cdx2 (or trophoblast stem cells) and Gata4 to reconstitute the epiblast, extraembryonic ectoderm and visceral endoderm lineages, respectively. The resulting complete embryo models recapitulate development from embryonic day 5.0 to 8.5, generating advanced embryonic and extraembryonic tissues that develop through gastrulation to initiate organogenesis to form a head and a beating heart structure as well as a yolk sac and chorion. Once the required stem cell lines are stably maintained in culture, the protocol requires 1 day to assemble complete embryo models and a further 8 days to culture them until headfold stages, although structures can be collected at earlier developmental stages as required. This protocol can be easily performed by researchers with experience in mouse stem cell culture, although they will benefit from knowledge of natural mouse embryos at early postimplantation stages.
", "doi": "10.1038/s41596-023-00891-y", "issn": "1754-2189", "publisher": "Nature Publishing Group", "publication": "Nature Protocols", "publication_date": "2023-12", "series_number": "12", "volume": "18", "issue": "12", "pages": "3662-3689" }, { "id": "authors:5bqk7-b1e47", "collection": "authors", "collection_id": "5bqk7-b1e47", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20230602-251540000.11", "type": "article", "title": "Stem cell-derived synthetic embryos self-assemble by exploiting cadherin codes and cortical tension", "author": [ { "family_name": "Bao", "given_name": "Min", "orcid": "0000-0003-0992-9388", "clpid": "Bao-Min" }, { "family_name": "Cornwall-Scoones", "given_name": "Jake", "orcid": "0000-0002-7435-486X", "clpid": "Cornwall-Scoones-Jake" }, { "family_name": "Sanchez-Vasquez", "given_name": "Estefania", "orcid": "0000-0002-6585-8548", "clpid": "Sanchez-Vasquez-Estefania" }, { "family_name": "Cox", "given_name": "Andy L.", "clpid": "Cox-Andy-L" }, { "family_name": "Chen", "given_name": "Dong-Yuan", "orcid": "0000-0003-2179-2847", "clpid": "Chen-Dong-Yuan" }, { "family_name": "De Jonghe", "given_name": "Joachim", "orcid": "0000-0003-0584-8265", "clpid": "De-Jonghe-Joachim" }, { "family_name": "Shadkhoo", "given_name": "Shahriar", "orcid": "0000-0003-3582-0634", "clpid": "Shadkhoo-Shahriar" }, { "family_name": "Hollfelder", "given_name": "Florian", "orcid": "0000-0002-1367-6312", "clpid": "Hollfelder-Florian" }, { "family_name": "Thomson", "given_name": "Matt", "orcid": "0000-0003-1021-1234", "clpid": "Thomson-M-W" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Mammalian embryos sequentially differentiate into trophectoderm and an inner cell mass, the latter of which differentiates into primitive endoderm and epiblast. Trophoblast stem (TS), extraembryonic endoderm (XEN) and embryonic stem (ES) cells derived from these three lineages can self-assemble into synthetic embryos, but the mechanisms remain unknown. Here, we show that a stem cell-specific cadherin code drives synthetic embryogenesis. The XEN cell cadherin code enables XEN cell sorting into a layer below ES cells, recapitulating the sorting of epiblast and primitive endoderm before implantation. The TS cell cadherin code enables TS cell sorting above ES cells, resembling extraembryonic ectoderm clustering above epiblast following implantation. Whereas differential cadherin expression drives initial cell sorting, cortical tension consolidates tissue organization. By optimizing cadherin code expression in different stem cell lines, we tripled the frequency of correctly formed synthetic embryos. Thus, by exploiting cadherin codes from different stages of development, lineage-specific stem cells bypass the preimplantation structure to directly assemble a postimplantation embryo.", "doi": "10.1038/s41556-022-00984-y", "pmcid": "PMC9481465", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2023-09", "series_number": "9", "volume": "24", "issue": "9", "pages": "1341-2349" }, { "id": "authors:8w05t-tbd96", "collection": "authors", "collection_id": "8w05t-tbd96", "cite_using_url": "https://authors.library.caltech.edu/records/8w05t-tbd96", "type": "article", "title": "spinDrop: a droplet microfluidic platform to maximise single-cell sequencing information content", "author": [ { "family_name": "De Jonghe", "given_name": "Joachim", "orcid": "0000-0003-0584-8265", "clpid": "De-Jonghe-Joachim" }, { "family_name": "Kaminski", "given_name": "Tomasz S.", "orcid": "0000-0001-5124-4548", "clpid": "Kaminski-Tomasz-S" }, { "family_name": "Morse", "given_name": "David B.", "clpid": "Morse-David-B" }, { "family_name": "Tabaka", "given_name": "Marcin", "orcid": "0000-0002-6166-2820", "clpid": "Tabaka-Marcin" }, { "family_name": "Ellermann", "given_name": "Anna L.", "orcid": "0000-0002-3713-2870", "clpid": "Ellermann-Anna-L" }, { "family_name": "Kohler", "given_name": "Timo N.", "orcid": "0000-0003-1949-0655", "clpid": "Kohler-Timo-N" }, { "family_name": "Amadei", "given_name": "Gianluca", "orcid": "0000-0001-5405-968X", "clpid": "Amadei-Gianluca" }, { "family_name": "Handford", "given_name": "Charlotte E.", "orcid": "0000-0002-5245-8027", "clpid": "Handford-Charlotte-E" }, { "family_name": "Findlay", "given_name": "Gregory M.", "orcid": "0000-0002-7767-8608", "clpid": "Findlay-Gregory-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Teichmann", "given_name": "Sarah A.", "orcid": "0000-0002-6294-6366", "clpid": "Teichmann-Sarah-A" }, { "family_name": "Hollfelder", "given_name": "Florian", "orcid": "0000-0002-1367-6312", "clpid": "Hollfelder-Florian" } ], "abstract": "Droplet microfluidic methods have massively increased the throughput of single-cell sequencing campaigns. The benefit of scale-up is, however, accompanied by increased background noise when processing challenging samples and the overall RNA capture efficiency is lower. These drawbacks stem from the lack of strategies to enrich for high-quality material or specific cell types at the moment of cell encapsulation and the absence of implementable multi-step enzymatic processes that increase capture. Here we alleviate both bottlenecks using fluorescence-activated droplet sorting to enrich for droplets that contain single viable cells, intact nuclei, fixed cells or target cell types and use reagent addition to droplets by picoinjection to perform multi-step lysis and reverse transcription. Our methodology increases gene detection rates fivefold, while reducing background noise by up to half. We harness these properties to deliver a high-quality molecular atlas of mouse brain development, despite starting with highly damaged input material, and provide an atlas of nascent RNA transcription during mouse organogenesis. Our method is broadly applicable to other droplet-based workflows to deliver sensitive and accurate single-cell profiling at a reduced cost.
", "doi": "10.1038/s41467-023-40322-w", "pmcid": "PMC10409775", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2023-08-08", "volume": "14", "pages": "4788" }, { "id": "authors:4ntb3-vv811", "collection": "authors", "collection_id": "4ntb3-vv811", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20230628-295417000.9", "type": "article", "title": "A model of the post-implantation human embryo derived from pluripotent stem cells", "author": [ { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278", "clpid": "Weatherbee-Bailey-A-T" }, { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786", "clpid": "Gantner-Carlos-W" }, { "family_name": "Iwamoto-Stohl", "given_name": "Lisa K.", "orcid": "0000-0003-0776-054X", "clpid": "Iwamoto-Stohl-Lisa-K" }, { "family_name": "Daza", "given_name": "Riza M.", "orcid": "0000-0003-1635-8675", "clpid": "Daza-Riza-M" }, { "family_name": "Hamazaki", "given_name": "Nobuhiko", "orcid": "0000-0001-9228-233X", "clpid": "Hamazaki-Nobuhiko" }, { "family_name": "Shendure", "given_name": "Jay", "orcid": "0000-0002-1516-1865", "clpid": "Shendure-Jay" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The human embryo undergoes morphogenetic transformations following implantation into the uterus, yet our knowledge of this crucial stage is limited by the inability to observe the embryo in vivo. Stem cell-derived models of the embryo are important tools to interrogate developmental events and tissue-tissue crosstalk during these stages. Here, we establish a model of the human post-implantation embryo, a human embryoid, comprised of embryonic and extraembryonic tissues. We combine two types of extraembryonic-like cells generated by transcription factor overexpression with wildtype embryonic stem cells and promote their self-organization into structures that mimic several aspects of the post-implantation human embryo. These self-organized aggregates contain a pluripotent epiblast-like domain surrounded by extraembryonic-like tissues. Our functional studies demonstrate that the epiblast-like domain robustly differentiates to amnion, extraembryonic mesenchyme, and primordial germ cell-like cells in response to BMP cues. In addition, we identify an inhibitory role for SOX17 in the specification of anterior hypoblast-like cells. Modulation of the subpopulations in the hypoblast-like compartment demonstrated that extraembryonic-like cells impact epiblast-like domain differentiation, highlighting functional tissue-tissue crosstalk. In conclusion, we present a modular, tractable, integrated model of the human embryo that will allow us to probe key questions of human post-implantation development, a critical window when significant numbers of pregnancies fail.", "doi": "10.1038/s41586-023-06368-y", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2023-06-28" }, { "id": "authors:at8q6-yzk07", "collection": "authors", "collection_id": "at8q6-yzk07", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20230629-647980000.1", "type": "monograph", "title": "Transgene directed induction of a stem cell-derived human embryo model", "author": [ { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278", "clpid": "Weatherbee-Bailey-A-T" }, { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786", "clpid": "Gantner-Carlos-W" }, { "family_name": "Daza", "given_name": "Riza M.", "orcid": "0000-0003-1635-8675", "clpid": "Daza-Riza-M" }, { "family_name": "Hamazaki", "given_name": "Nobuhiko", "orcid": "0000-0001-9228-233X", "clpid": "Hamazaki-Nobuhiko" }, { "family_name": "Iwamoto-Stohl", "given_name": "Lisa K.", "orcid": "0000-0003-0776-054X", "clpid": "Iwamoto-Stohl-Lisa-K" }, { "family_name": "Shendure", "given_name": "Jay", "orcid": "0000-0002-1516-1865", "clpid": "Shendure-Jay" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The human embryo undergoes morphogenetic transformations following implantation into the uterus, yet our knowledge of this crucial stage is limited by the inability to observe the embryo in vivo. Stem cell-derived models of the embryo are important tools to interrogate developmental events and tissue-tissue crosstalk during these stages. Here, we establish a human post-implantation embryo model comprised of embryonic and extraembryonic tissues. We combine two types of extraembryonic-like cells generated by transcription factor overexpression with wildtype embryonic stem cells and promote their self-organization into structures that mimic aspects of the post-implantation human embryo. These self-organized aggregates contain a pluripotent epiblast-like domain surrounded by hypoblast-and trophoblast-like tissues. We demonstrate that these inducible human embryoids robustly generate several cell types, including amnion, extraembryonic mesenchyme, and primordial germ cell-like cells in response to BMP signaling. This model also allowed us to identify an inhibitory role for SOX17 in the specification of anterior hypoblast-like cells. Modulation of the subpopulations in the hypoblast-like compartment demonstrated that extraembryonic-like cells impact epiblast-like domain differentiation, highlighting functional tissue-tissue crosstalk. In conclusion, we present a modular, tractable, integrated model of the human embryo that will allow us to probe key questions of human post-implantation development, a critical window when significant numbers of pregnancies fail.", "doi": "10.1101/2023.06.15.545082", "publication_date": "2023-06-15" }, { "id": "authors:x6fp2-2w646", "collection": "authors", "collection_id": "x6fp2-2w646", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20230725-857148000.45", "type": "article", "title": "Progress and challenges in stem cell biology", "author": [ { "family_name": "Apostolou", "given_name": "Effie", "orcid": "0000-0002-8111-0863", "clpid": "Apostolou-Effie" }, { "family_name": "Blau", "given_name": "Helen", "orcid": "0000-0001-6503-5480", "clpid": "Blau-Helen" }, { "family_name": "Chien", "given_name": "Kenneth", "orcid": "0000-0002-2759-8378", "clpid": "Chien-Kenneth" }, { "family_name": "Lancaster", "given_name": "Madeline A.", "orcid": "0000-0003-2324-8853", "clpid": "Lancaster-Madeline-A" }, { "family_name": "Tata", "given_name": "Purushothama Rao", "orcid": "0000-0003-4837-0337", "clpid": "Tata-Purushothama-Rao" }, { "family_name": "Trompouki", "given_name": "Eirini", "orcid": "0000-0002-7242-8810", "clpid": "Trompouki-Eirini" }, { "family_name": "Watt", "given_name": "Fiona M.", "orcid": "0000-0001-9151-5154", "clpid": "Watt-Fiona-M" }, { "family_name": "Zeng", "given_name": "Yi Arial", "orcid": "0000-0003-1898-8099", "clpid": "Zeng-Yi-Arial" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Since stem cells were first discovered, researchers have identified distinct stem cell populations in different organs and with various functions, converging on the unique abilities of self-renewal and differentiation toward diverse cell types. These abilities make stem cells an incredibly promising tool in therapeutics and have turned stem cell biology into a fast-evolving field. Here, stem cell biologists express their view on the most striking advances and current challenges in their field.", "doi": "10.1038/s41556-023-01087-y", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2023-02", "series_number": "2", "volume": "25", "issue": "2", "pages": "203-206" }, { "id": "authors:vvfn6-skp75", "collection": "authors", "collection_id": "vvfn6-skp75", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20230316-182577000.43", "type": "monograph", "title": "spinDrop: a droplet microfluidic platform to maximise single-cell sequencing information content", "author": [ { "family_name": "De Jonghe", "given_name": "Joachim", "orcid": "0000-0003-0584-8265", "clpid": "De-Jonghe-Joachim" }, { "family_name": "Kaminski", "given_name": "Tomasz S.", "orcid": "0000-0001-5124-4548", "clpid": "Kaminski-Tomasz-S" }, { "family_name": "Morse", "given_name": "David B.", "clpid": "Morse-David-B" }, { "family_name": "Tabaka", "given_name": "Marcin", "orcid": "0000-0002-6166-2820", "clpid": "Tabaka-Marcin" }, { "family_name": "Ellermann", "given_name": "Anna L.", "clpid": "Ellermann-Anna-L" }, { "family_name": "Kohler", "given_name": "Timo N.", "orcid": "0000-0003-1949-0655", "clpid": "Kohler-Timo-N" }, { "family_name": "Amadei", "given_name": "Gianluca", "orcid": "0000-0001-5405-968X", "clpid": "Amadei-Gianluca" }, { "family_name": "Handford", "given_name": "Charlotte", "orcid": "0000-0002-5245-8027", "clpid": "Handford-Charlotte-E" }, { "family_name": "Findlay", "given_name": "Gregory M.", "clpid": "Findlay-Gregory-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Teichmann", "given_name": "Sarah A.", "orcid": "0000-0002-6294-6366", "clpid": "Teichmann-Sarah-A" }, { "family_name": "Hollfelder", "given_name": "Florian", "orcid": "0000-0002-1367-6312", "clpid": "Hollfelder-Florian" } ], "abstract": "Droplet microfluidic methods have massively increased the throughput of single-cell sequencing campaigns. The benefit of scale-up is, however, accompanied by increased background noise when processing challenging samples and the overall RNA capture efficiency is lower. These drawbacks stem from the lack of strategies to enrich for high-quality material or specific cell types at the moment of cell encapsulation and the absence of implementable multi-step enzymatic processes that increase capture. Here we alleviate both bottlenecks using fluorescence-activated droplet sorting to enrich for droplets that contain single viable cells, intact nuclei, fixed cells or target cell types and use reagent addition to droplets by picoinjection to perform multi-step lysis and reverse transcription. Our methodology increases gene detection rates fivefold, while reducing background noise by up to half. We harness these unique properties to deliver a high-quality molecular atlas of mouse brain development, despite starting with highly damaged input material, and provide an atlas of nascent RNA transcription during mouse organogenesis. Our method is broadly applicable to other droplet-based workflows to deliver sensitive and accurate single-cell profiling at a reduced cost.", "doi": "10.1101/2023.01.12.523500", "publication_date": "2023-01-14" }, { "id": "authors:b2j0f-2xk53", "collection": "authors", "collection_id": "b2j0f-2xk53", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20221122-564647900.8", "type": "article", "title": "Mouse embryo model derived exclusively from embryonic stem cells undergoes neurulation and heart development", "author": [ { "family_name": "Lau", "given_name": "Kasey Y. C.", "clpid": "Lau-Kasey-Y-C" }, { "family_name": "Rubinstein", "given_name": "Hernan", "clpid": "Rubinstein-Hernan" }, { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786", "clpid": "Gantner-Carlos-W" }, { "family_name": "Hadas", "given_name": "Ron", "orcid": "0000-0001-9301-6607", "clpid": "Hadas-Ron" }, { "family_name": "Amadei", "given_name": "Gianluca", "orcid": "0000-0001-5405-968X", "clpid": "Amadei-Gianluca" }, { "family_name": "Stelzer", "given_name": "Yonatan", "orcid": "0000-0001-9207-1479", "clpid": "Stelzer-Yonatan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Several in vitro models have been developed to recapitulate mouse embryogenesis solely from embryonic stem cells (ESCs). Despite mimicking many aspects of early development, they fail to capture the interactions between embryonic and extraembryonic tissues. To overcome this difficulty, we have developed a mouse ESC-based in vitro model that reconstitutes the pluripotent ESC lineage and the two extraembryonic lineages of the post-implantation embryo by transcription-factor-mediated induction. This unified model recapitulates developmental events from embryonic day 5.5 to 8.5, including gastrulation; formation of the anterior-posterior axis, brain, and a beating heart structure; and the development of extraembryonic tissues, including yolk sac and chorion. Comparing single-cell RNA sequencing from individual structures with time-matched natural embryos identified remarkably similar transcriptional programs across lineages but also showed when and where the model diverges from the natural program. Our findings demonstrate an extraordinary plasticity of ESCs to self-organize and generate a whole-embryo-like structure.", "doi": "10.1016/j.stem.2022.08.013", "pmcid": "PMC9648694", "issn": "1934-5909", "publisher": "Cell Press", "publication": "Cell Stem Cell", "publication_date": "2022-10-06", "series_number": "10", "volume": "29", "issue": "10", "pages": "1445-1458.e8" }, { "id": "authors:ykz2r-3p577", "collection": "authors", "collection_id": "ykz2r-3p577", "cite_using_url": "https://authors.library.caltech.edu/records/ykz2r-3p577", "type": "article", "title": "Mouse embryo model derived exclusively from embryonic stem cells undergoes neurulation and heart development", "author": [ { "family_name": "Lau", "given_name": "Kasey Y. C.", "clpid": "Lau-Kasey-Y-C" }, { "family_name": "Rubinstein", "given_name": "Hernan", "clpid": "Rubinstein-Hernan" }, { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786", "clpid": "Gantner-Carlos-W" }, { "family_name": "Hadas", "given_name": "Ron", "orcid": "0000-0001-9301-6607", "clpid": "Hadas-Ron" }, { "family_name": "Amadei", "given_name": "Gianluca" }, { "family_name": "Stelzer", "given_name": "Yonatan", "orcid": "0000-0001-9207-1479", "clpid": "Stelzer-Yonatan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Several in vitro models have been developed to recapitulate mouse embryogenesis solely from embryonic stem cells (ESCs). Despite mimicking many aspects of early development, they fail to capture the interactions between embryonic and extraembryonic tissues. To overcome this difficulty, we have developed a mouse ESC-based in vitro model that reconstitutes the pluripotent ESC lineage and the two extraembryonic lineages of the post-implantation embryo by transcription-factor-mediated induction. This unified model recapitulates developmental events from embryonic day 5.5 to 8.5, including gastrulation; formation of the anterior-posterior axis, brain, and a beating heart structure; and the development of extraembryonic tissues, including yolk sac and chorion. Comparing single-cell RNA sequencing from individual structures with time-matched natural embryos identified remarkably similar transcriptional programs across lineages but also showed when and where the model diverges from the natural program. Our findings demonstrate an extraordinary plasticity of ESCs to self-organize and generate a whole-embryo-like structure.
", "doi": "10.1016/j.stem.2022.08.013", "pmcid": "PMC9648694", "issn": "1934-5909", "publisher": "Cell Press", "publication": "Cell Stem Cell", "publication_date": "2022-10-06", "series_number": "10", "volume": "29", "issue": "10", "pages": "1445-1458.e8" }, { "id": "authors:ny5rx-x3886", "collection": "authors", "collection_id": "ny5rx-x3886", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20221010-454096500.25", "type": "article", "title": "Embryo\u00a0model completes gastrulation to neurulation and organogenesis", "author": [ { "family_name": "Amadei", "given_name": "Gianluca", "orcid": "0000-0001-5405-968X", "clpid": "Amadei-Gianluca" }, { "family_name": "Handford", "given_name": "Charlotte E.", "orcid": "0000-0002-5245-8027", "clpid": "Handford-Charlotte-E" }, { "family_name": "Qiu", "given_name": "Chengxiang", "orcid": "0000-0002-6346-8669", "clpid": "Qiu-Chengxiang" }, { "family_name": "De Jonghe", "given_name": "Joachim", "orcid": "0000-0003-0584-8265", "clpid": "De-Jonghe-Joachim" }, { "family_name": "Greenfeld", "given_name": "Hannah", "clpid": "Greenfeld-Hannah" }, { "family_name": "Tran", "given_name": "Martin", "clpid": "Tran-Martin" }, { "family_name": "Martin", "given_name": "Beth K.", "orcid": "0000-0002-9661-014X", "clpid": "Martin-Beth-K" }, { "family_name": "Chen", "given_name": "Dong-Yuan", "orcid": "0000-0003-2179-2847", "clpid": "Chen-Dong-Yuan" }, { "family_name": "Aguilera-Castrejon", "given_name": "Alejandro", "orcid": "0000-0002-1339-7778", "clpid": "Aguilera-Castrejon-Alejandro" }, { "family_name": "Hanna", "given_name": "Jacob H.", "orcid": "0000-0003-2042-9974", "clpid": "Hanna-Jacob-H" }, { "family_name": "Elowitz", "given_name": "Michael B.", "orcid": "0000-0002-1221-0967", "clpid": "Elowitz-M-B" }, { "family_name": "Hollfelder", "given_name": "Florian", "orcid": "0000-0002-1367-6312", "clpid": "Hollfelder-Florian" }, { "family_name": "Shendure", "given_name": "Jay", "orcid": "0000-0002-1516-1865", "clpid": "Shendure-Jay" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Embryonic stem (ES) cells can undergo many aspects of mammalian embryogenesis in vitro, but their developmental potential is substantially extended by interactions with extraembryonic stem cells, including trophoblast stem (TS) cells, extraembryonic endoderm stem (XEN) cells and inducible XEN (iXEN) cells. Here we assembled stem cell-derived embryos in vitro from mouse ES cells, TS cells and iXEN cells and showed that they recapitulate the development of whole natural mouse embryo in utero up to day 8.5 post-fertilization. Our embryo model displays headfolds with defined forebrain and midbrain regions and develops a beating heart-like structure, a trunk comprising a neural tube and somites, a tail bud containing neuromesodermal progenitors, a gut tube, and primordial germ cells. This complete embryo model develops within an extraembryonic yolk sac that initiates blood island development. Notably, we demonstrate that the neurulating embryo model assembled from Pax6-knockout ES cells aggregated with wild-type TS cells and iXEN cells recapitulates the ventral domain expansion of the neural tube that occurs in natural, ubiquitous Pax6-knockout embryos. Thus, these complete embryoids are a powerful in vitro model for dissecting the roles of diverse cell lineages and genes in development. Our results demonstrate the self-organization ability of ES cells and two types of extraembryonic stem cells to reconstitute mammalian development through and beyond gastrulation to neurulation and early organogenesis.", "doi": "10.1038/s41586-022-05246-3", "pmcid": "PMC9534772", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2022-10-06", "series_number": "7930", "volume": "610", "issue": "7930", "pages": "143-153" }, { "id": "authors:78vb2-pyc16", "collection": "authors", "collection_id": "78vb2-pyc16", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220909-232668000", "type": "article", "title": "Stem-cell-based human and mouse embryo models", "author": [ { "family_name": "Bao", "given_name": "Min", "orcid": "0000-0003-0992-9388", "clpid": "Bao-Min" }, { "family_name": "Cornwall-Scoones", "given_name": "Jake", "orcid": "0000-0002-7435-486X", "clpid": "Cornwall-Scoones-Jake" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Synthetic embryology aims to develop embryo-like structures from stem cells to provide new insight into early stages of mammalian development. Recent advances in synthetic embryology have highlighted the remarkable capacity of stem cells to self-organize under certain biochemical or biophysical stimulations, generating structures that recapitulate the fate and form of early mouse/human embryos, in which symmetry breaking, pattern formation, or proper morphogenesis can be observed spontaneously. Here we review recent progress on the design principles for different types of embryoids and discuss the impact of different biochemical and biophysical factors on the process of stem-cell self-organization. We also offer our thoughts about the principal future challenges.", "doi": "10.1016/j.gde.2022.101970", "pmcid": "PMC10309046", "issn": "0959-437X", "publisher": "Elsevier", "publication": "Current Opinion in Genetics and Development", "publication_date": "2022-10", "volume": "76", "pages": "101970" }, { "id": "authors:am7bp-3g465", "collection": "authors", "collection_id": "am7bp-3g465", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220919-149101800", "type": "article", "title": "Stem cell-derived synthetic embryos self-assemble by exploiting cadherin codes and cortical tension", "author": [ { "family_name": "Bao", "given_name": "Min", "orcid": "0000-0003-0992-9388", "clpid": "Bao-Min" }, { "family_name": "Cornwall-Scoones", "given_name": "Jake", "orcid": "0000-0002-7435-486X", "clpid": "Cornwall-Scoones-Jake" }, { "family_name": "Sanchez-Vasquez", "given_name": "Estefania", "orcid": "0000-0002-6585-8548", "clpid": "Sanchez-Vasquez-Estefania" }, { "family_name": "Cox", "given_name": "Andy L.", "clpid": "Cox-Andy-L" }, { "family_name": "Chen", "given_name": "Dong-Yuan", "orcid": "0000-0003-2179-2847", "clpid": "Chen-Dong-Yuan" }, { "family_name": "De Jonghe", "given_name": "Joachim", "orcid": "0000-0003-0584-8265", "clpid": "De-Jonghe-Joachim" }, { "family_name": "Shadkhoo", "given_name": "Shahriar", "orcid": "0000-0003-3582-0634", "clpid": "Shadkhoo-Shahriar" }, { "family_name": "Hollfelder", "given_name": "Florian", "orcid": "0000-0002-1367-6312", "clpid": "Hollfelder-Florian" }, { "family_name": "Thomson", "given_name": "Matt", "orcid": "0000-0003-1021-1234", "clpid": "Thomson-M-W" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Mammalian embryos sequentially differentiate into trophectoderm and an inner cell mass, the latter of which differentiates into primitive endoderm and epiblast. Trophoblast stem (TS), extraembryonic endoderm (XEN) and embryonic stem (ES) cells derived from these three lineages can self-assemble into synthetic embryos, but the mechanisms remain unknown. Here, we show that a stem cell-specific cadherin code drives synthetic embryogenesis. The XEN cell cadherin code enables XEN cell sorting into a layer below ES cells, recapitulating the sorting of epiblast and primitive endoderm before implantation. The TS cell cadherin code enables TS cell sorting above ES cells, resembling extraembryonic ectoderm clustering above epiblast following implantation. Whereas differential cadherin expression drives initial cell sorting, cortical tension consolidates tissue organization. By optimizing cadherin code expression in different stem cell lines, we tripled the frequency of correctly formed synthetic embryos. Thus, by exploiting cadherin codes from different stages of development, lineage-specific stem cells bypass the preimplantation structure to directly assemble a postimplantation embryo.", "doi": "10.1038/s41556-022-00984-y", "pmcid": "PMC9481465", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2022-09-13", "series_number": "9", "volume": "24", "issue": "9", "pages": "1341-1349" }, { "id": "authors:3tv59-q5m26", "collection": "authors", "collection_id": "3tv59-q5m26", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220826-428674800.766", "type": "article", "title": "E-cadherin mediates apical membrane initiation site localisation during de\u2009novo polarisation of epithelial cavities", "author": [ { "family_name": "Liang", "given_name": "Xuan", "orcid": "0000-0003-4019-1457", "clpid": "Liang-Xuan" }, { "family_name": "Weberling", "given_name": "Antonia", "orcid": "0000-0001-8282-5695", "clpid": "Weberling-Antonia" }, { "family_name": "Hii", "given_name": "Chun Yuan", "orcid": "0000-0002-5344-9747", "clpid": "Hii-Chun-Yuan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Buckley", "given_name": "Clare E.", "orcid": "0000-0003-3329-3973", "clpid": "Buckley-Clare-E" } ], "abstract": "Individual cells within de novo polarising tubes and cavities must integrate their forming apical domains into a centralised apical membrane initiation site (AMIS). This is necessary to enable organised lumen formation within multi-cellular tissue. Despite the well-documented importance of cell division in localising the AMIS, we have found a division-independent mechanism of AMIS localisation that relies instead on Cadherin-mediated cell\u2013cell adhesion. Our study of de novo polarising mouse embryonic stem cells (mESCs) cultured in 3D suggests that cell\u2013cell adhesion localises apical proteins such as PAR-6 to a centralised AMIS. Unexpectedly, we also found that mESC clusters lacking functional E-cadherin still formed a lumen-like cavity in the absence of AMIS localisation but did so at a later stage of development via a \"closure\" mechanism, instead of via hollowing. This work suggests that there are two, interrelated mechanisms of apical polarity localisation: cell adhesion and cell division. Alignment of these mechanisms in space allows for redundancy in the system and ensures the development of a coherent epithelial structure within a growing organ.", "doi": "10.15252/embj.2022111021", "issn": "0261-4189", "publisher": "European Molecular Biology Organization", "publication": "EMBO Journal", "publication_date": "2022-08-31", "pages": "Art No. e111021" }, { "id": "authors:z18s9-5vg81", "collection": "authors", "collection_id": "z18s9-5vg81", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220803-224511000", "type": "monograph", "title": "Mouse-embryo model derived exclusively from embryonic stem cells undergo neurulation and heart development", "author": [ { "family_name": "Lau", "given_name": "Kasey Y. C.", "clpid": "Lau-Kasey-Y-C" }, { "family_name": "Rubinstein", "given_name": "Hernan", "clpid": "Rubinstein-Hernan" }, { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786", "clpid": "Gantner-Carlos-W" }, { "family_name": "Amadei", "given_name": "Gianluca", "orcid": "0000-0001-5405-968X", "clpid": "Amadei-Gianluca" }, { "family_name": "Stelzer", "given_name": "Yonatan", "orcid": "0000-0001-9207-1479", "clpid": "Stelzer-Yonatan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Several in vitro models have been developed to recapitulate mouse embryogenesis solely from embryonic stem cells (ESCs). Despite mimicking many aspects of early development, they fail to capture the interactions between embryonic and extraembryonic tissues. To overcome this difficulty, we have developed a mouse ESC-based in vitro model that reconstitutes the pluripotent ESC lineage and the two extra-embryonic lineages of the post-implantation embryo by transcription factor-mediated induction. This unified model recapitulates developmental events from embryonic day 5.5 to 8.5, including gastrulation, and formation of the anterior-posterior axis, brain, a beating heart structure, and the development of extraembryonic tissues, including yolk sac and chorion. Comparing single-cell RNA sequencing from individual structures with time-matched natural embryos identified remarkably similar transcriptional programs across lineages, but also showed when and where the model diverges from the natural program. Our findings demonstrate an extra-ordinary plasticity of ESCs to self-organize and generate a whole embryo-like structure.", "doi": "10.1101/2022.08.01.502371", "publication_date": "2022-08-02" }, { "id": "authors:1g1ew-0ne83", "collection": "authors", "collection_id": "1g1ew-0ne83", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220803-224696000", "type": "monograph", "title": "Stem cell-derived mouse embryos develop within an extra-embryonic yolk sac to form anterior brain regions and a beating heart", "author": [ { "family_name": "Amadei", "given_name": "Gianluca", "clpid": "Amadei-Gianluca" }, { "family_name": "Handford", "given_name": "Charlotte E.", "clpid": "Handford-Charlotte-E" }, { "family_name": "De Jonghe", "given_name": "Joachim", "clpid": "De-Jonghe-Joachim" }, { "family_name": "Hollfelder", "given_name": "Florian", "orcid": "0000-0002-1367-6312", "clpid": "Hollfelder-Florian" }, { "family_name": "Glover", "given_name": "David", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Embryo-like structures generated from stem cells can achieve varying developmental milestones, but none have been shown to progress through gastrulation, neurulation, and organogenesis. Here, we show that ETiX mouse embryos, established from embryonic stem cells aggregated with trophoblast stem cells and inducible extraembryonic endoderm stem cells, can develop through gastrulation and beyond to undertake neural induction and generate the progenitors needed to create the entire organism. The head-folds of ETiX embryos show anterior expression of Otx2, defining forebrain and midbrain regions that resemble those of the natural mouse embryo. ETiX embryos also develop beating hearts, trunk structures comprising a neural tube and somites, tail buds containing neuromesodermal progenitors and primordial germ cells, and gut tubes derived from definitive endoderm. A fraction of ETiX embryos show neural tube abnormalities, which can be partially rescued by treatment with the metabolically active form of folic acid, reminiscent of common birth defects and therapies in humans. Notably, ETiX embryos also develop a yolk sac with blood islands. Overall, ETiX embryos uniquely recapitulate natural embryos, developing further than any other stem-cell derived model, through multiple post-implantation stages and within extra-embryonic membranes.", "doi": "10.1101/2022.08.01.502375", "publication_date": "2022-08-02" }, { "id": "authors:k7p4y-d6485", "collection": "authors", "collection_id": "k7p4y-d6485", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220215-496417800", "type": "article", "title": "Stain-free detection of embryo polarization using deep learning", "author": [ { "family_name": "Shen", "given_name": "Cheng", "orcid": "0000-0001-7136-4715", "clpid": "Shen-Cheng" }, { "family_name": "Lamba", "given_name": "Adiyant", "clpid": "Lamba-Adiyant" }, { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Zhang", "given_name": "Ray", "clpid": "Zhang-Ray" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Yang", "given_name": "Changhuei", "orcid": "0000-0001-8791-0354", "clpid": "Yang-Changhuei" } ], "abstract": "Polarization of the mammalian embryo at the right developmental time is critical for its development to term and would be valuable in assessing the potential of human embryos. However, tracking polarization requires invasive fluorescence staining, impermissible in the in vitro fertilization clinic. Here, we report the use of artificial intelligence to detect polarization from unstained time-lapse movies of mouse embryos. We assembled a dataset of bright-field movie frames from 8-cell-stage embryos, side-by-side with corresponding images of fluorescent markers of cell polarization. We then used an ensemble learning model to detect whether any bright-field frame showed an embryo before or after onset of polarization. Our resulting model has an accuracy of 85% for detecting polarization, significantly outperforming human volunteers trained on the same data (61% accuracy). We discovered that our self-learning model focuses upon the angle between cells as one known cue for compaction, which precedes polarization, but it outperforms the use of this cue alone. By compressing three-dimensional time-lapsed image data into two-dimensions, we are able to reduce data to an easily manageable size for deep learning processing. In conclusion, we describe a method for detecting a key developmental feature of embryo development that avoids clinically impermissible fluorescence staining.", "doi": "10.1038/s41598-022-05990-6", "issn": "2045-2322", "publisher": "Nature Publishing Group", "publication": "Scientific Reports", "publication_date": "2022-02-14", "volume": "12", "pages": "Art. No. 2404" }, { "id": "authors:5pbcs-yw564", "collection": "authors", "collection_id": "5pbcs-yw564", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20220201-160162400", "type": "article", "title": "Lima1 mediates the pluripotency control of membrane dynamics and cellular metabolism", "author": [ { "family_name": "Duethorn", "given_name": "Binyamin", "clpid": "Duethorn-Binyamin" }, { "family_name": "Groll", "given_name": "Fabian", "clpid": "Groll-Fabian" }, { "family_name": "Rieger", "given_name": "Bettina", "clpid": "Rieger-Bettina" }, { "family_name": "Drexler", "given_name": "Hannes C. A.", "orcid": "0000-0002-8285-0644", "clpid": "Drexler-Hannes-C-A" }, { "family_name": "Brinkmann", "given_name": "Heike", "clpid": "Brinkmann-Heike" }, { "family_name": "Kremer", "given_name": "Ludmila", "clpid": "Kremer-Ludmila" }, { "family_name": "Stehling", "given_name": "Martin", "clpid": "Stehling-Martin" }, { "family_name": "Borowski", "given_name": "Marie-Theres", "orcid": "0000-0003-3648-7757", "clpid": "Borowski-Marie-Theres" }, { "family_name": "Mildner", "given_name": "Karina", "orcid": "0000-0002-6229-6519", "clpid": "Mildner-Karina" }, { "family_name": "Zeuschner", "given_name": "Dagmar", "orcid": "0000-0002-6712-0192", "clpid": "Zeuschner-Dagmar" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Stemmler", "given_name": "Marc P.", "orcid": "0000-0002-7866-3686", "clpid": "Stemmler-Marc-P" }, { "family_name": "Busch", "given_name": "Karin B.", "orcid": "0000-0003-0525-0191", "clpid": "Busch-Karin-B" }, { "family_name": "Vaquerizas", "given_name": "Juan M.", "orcid": "0000-0002-6583-6541", "clpid": "Vaquerizas-Juan-M" }, { "family_name": "Bedzhov", "given_name": "Ivan", "orcid": "0000-0002-2122-6485", "clpid": "Bedzhov-Ivan" } ], "abstract": "Lima1 is an extensively studied prognostic marker of malignancy and is also considered to be a tumour suppressor, but its role in a developmental context of non-transformed cells is poorly understood. Here, we characterise the expression pattern and examined the function of Lima1 in mouse embryos and pluripotent stem cell lines. We identify that Lima1 expression is controlled by the na\u00efve pluripotency circuit and is required for the suppression of membrane blebbing, as well as for proper mitochondrial energetics in embryonic stem cells. Moreover, forcing Lima1 expression enables primed mouse and human pluripotent stem cells to be incorporated into murine pre-implantation embryos. Thus, Lima1 is a key effector molecule that mediates the pluripotency control of membrane dynamics and cellular metabolism.", "doi": "10.1038/s41467-022-28139-5", "pmcid": "PMC8807836", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2022-02-01", "volume": "13", "pages": "Art. No. 610" }, { "id": "authors:t2m9x-pbn24", "collection": "authors", "collection_id": "t2m9x-pbn24", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20211214-786549900", "type": "monograph", "title": "E-cadherin mediated Apical Membrane Initiation Site localisation", "author": [ { "family_name": "Liang", "given_name": "Xuan", "orcid": "0000-0003-4019-1457", "clpid": "Liang-Xuan" }, { "family_name": "Weberling", "given_name": "Antonia", "orcid": "0000-0001-8282-5695", "clpid": "Weberling-Antonia" }, { "family_name": "Hii", "given_name": "Chun Yuan", "orcid": "0000-0002-5344-9747", "clpid": "Hii-Chun-Yuan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Buckley", "given_name": "Clare E.", "orcid": "0000-0003-3329-3973", "clpid": "Buckley-Clare-E" } ], "abstract": "Individual cells within de novo polarising tubes and cavities must integrate their forming apical domains into a centralised apical membrane initiation site (AMIS). This is necessary to enable organised lumen formation within multi-cellular tissue. Despite the well documented importance of cell division in localising the AMIS, we have found a division-independent mechanism of AMIS localisation that relies instead on Cadherin-mediated cell-cell adhesion. Our study of de novo polarising mouse embryonic stem cells (mESCs) cultured in 3D suggest that cell-cell adhesion localises apical proteins such as PAR-6 to a centralised AMIS. Unexpectedly, we also found that mESC cell clusters lacking functional E-cadherin still formed a lumen-like cavity in the absence of AMIS localisation but did so at a later stage of development via a closure mechanism, instead of via hollowing. This work suggests that there are two, interrelated mechanisms of apical polarity localisation: cell adhesion and cell division. Alignment of these mechanisms in space allows for redundancy in the system and ensures the development of a coherent epithelial structure within a growing organ.", "doi": "10.1101/2021.11.30.470571", "publication_date": "2021-12-02" }, { "id": "authors:ews1q-6g437", "collection": "authors", "collection_id": "ews1q-6g437", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20211124-161127351", "type": "article", "title": "Modelling the impact of decidual senescence on embryo implantation in human endometrial assembloids", "author": [ { "family_name": "Rawlings", "given_name": "Thomas M.", "clpid": "Rawlings-Thomas-M" }, { "family_name": "Makwana", "given_name": "Komal", "clpid": "Makwana-Komal" }, { "family_name": "Taylor", "given_name": "Deborah M.", "clpid": "Taylor-Deborah-M" }, { "family_name": "Mol\u00e8", "given_name": "Matteo A.", "orcid": "0000-0001-7342-4849", "clpid": "Mol\u00e8-Matteo-A" }, { "family_name": "Fishwick", "given_name": "Katherine J.", "clpid": "Fishwick-Katherine-J" }, { "family_name": "Tryfonos", "given_name": "Maria", "clpid": "Tryfonos-Maria" }, { "family_name": "Odendaal", "given_name": "Joshua", "orcid": "0000-0001-7829-3729", "clpid": "Odendaal-Joshua" }, { "family_name": "Hawkes", "given_name": "Amelia", "clpid": "Hawkes-Amelia" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Hartshorne", "given_name": "Geraldine M.", "clpid": "Hartshorne-Gerladine-M" }, { "family_name": "Brosens", "given_name": "Jan J.", "orcid": "0000-0003-0116-9329", "clpid": "Brosens-Jan-J" }, { "family_name": "Lucas", "given_name": "Emma S.", "orcid": "0000-0002-8571-8921", "clpid": "Lucas-Emma-S" } ], "abstract": "Decidual remodelling of midluteal endometrium leads to a short implantation window after which the uterine mucosa either breaks down or is transformed into a robust matrix that accommodates the placenta throughout pregnancy. To gain insights into the underlying mechanisms, we established and characterized endometrial assembloids, consisting of gland-like organoids and primary stromal cells. Single-cell transcriptomics revealed that decidualized assembloids closely resemble midluteal endometrium, harbouring differentiated and senescent subpopulations in both glands and stroma. We show that acute senescence in glandular epithelium drives secretion of multiple canonical implantation factors, whereas in the stroma it calibrates the emergence of anti-inflammatory decidual cells and pro-inflammatory senescent decidual cells. Pharmacological inhibition of stress responses in pre-decidual cells accelerated decidualization by eliminating the emergence of senescent decidual cells. In co-culture experiments, accelerated decidualization resulted in entrapment of collapsed human blastocysts in a robust, static decidual matrix. By contrast, the presence of senescent decidual cells created a dynamic implantation environment, enabling embryo expansion and attachment, although their persistence led to gradual disintegration of assembloids. Our findings suggest that decidual senescence controls endometrial fate decisions at implantation and highlight how endometrial assembloids may accelerate the discovery of new treatments to prevent reproductive failure.", "doi": "10.7554/elife.69603", "pmcid": "PMC8523170", "issn": "2050-084X", "publisher": "eLife Sciences Publications", "publication": "eLife", "publication_date": "2021-10-18", "volume": "10", "pages": "Art. No. e69603" }, { "id": "authors:k7txd-cgj35", "collection": "authors", "collection_id": "k7txd-cgj35", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200925-104734056", "type": "article", "title": "Human embryo polarization requires PLC signaling to mediate trophectoderm specification", "author": [ { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Shahbazi", "given_name": "Marta", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-Marta-N" }, { "family_name": "Martin", "given_name": "Angel", "clpid": "Martin-Angel" }, { "family_name": "Zhang", "given_name": "Chuanxin", "orcid": "0000-0003-0814-9275", "clpid": "Zhang-Chuanxin" }, { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-Berna" }, { "family_name": "Borsos", "given_name": "Mate", "orcid": "0000-0002-2801-8910", "clpid": "Borsos-M\u00e1t\u00e9" }, { "family_name": "Mandelbaum", "given_name": "Rachel S.", "orcid": "0000-0001-8611-1462", "clpid": "Mandelbaum-Rachel-S" }, { "family_name": "Paulson", "given_name": "Richard J.", "clpid": "Paulson-Richard-J" }, { "family_name": "Mol\u00e8", "given_name": "Matteo A.", "clpid": "Mol\u00e8-Matteo-A" }, { "family_name": "Esbert", "given_name": "Marga", "orcid": "0000-0002-0002-2272", "clpid": "Esbert-Marga" }, { "family_name": "Titus", "given_name": "Shiny", "orcid": "0000-0002-8391-825X", "clpid": "Titus-Shiny" }, { "family_name": "Scott", "given_name": "Richard T.", "clpid": "Scott-Richard-T-W" }, { "family_name": "Campbell", "given_name": "Alison", "orcid": "0000-0002-4065-5672", "clpid": "Campbell-Alison" }, { "family_name": "Fishel", "given_name": "Simon", "clpid": "Fishel-Simon" }, { "family_name": "Gradinaru", "given_name": "Viviana", "orcid": "0000-0001-5868-348X", "clpid": "Gradinaru-V" }, { "family_name": "Zhao", "given_name": "Han", "clpid": "Zhao-Han" }, { "family_name": "Wu", "given_name": "Keliang", "clpid": "Wu-Keliang" }, { "family_name": "Chen", "given_name": "Zi-Jiang", "clpid": "Chen-Zi-Jiang" }, { "family_name": "Seli", "given_name": "Emre", "orcid": "0000-0001-7464-8203", "clpid": "Seli-Emre" }, { "family_name": "de los Santos", "given_name": "Maria J.", "clpid": "de-los-Santos-Maria-Jose" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Apico-basal polarization of cells within the embryo is critical for the segregation of distinct lineages during mammalian development. Polarized cells become the trophectoderm (TE), which forms the placenta, and apolar cells become the inner cell mass (ICM), the founding population of the fetus. The cellular and molecular mechanisms leading to polarization of the human embryo and its timing during embryogenesis have remained unknown. Here, we show that human embryo polarization occurs in two steps: it begins with the apical enrichment of F-actin and is followed by the apical accumulation of the PAR complex. This two-step polarization process leads to the formation of an apical domain at the 8\u201316 cell stage. Using RNA interference, we show that apical domain formation requires Phospholipase C (PLC) signaling, specifically the enzymes PLCB1 and PLCE1, from the eight-cell stage onwards. Finally, we show that although expression of the critical TE differentiation marker GATA3 can be initiated independently of embryo polarization, downregulation of PLCB1 and PLCE1 decreases GATA3 expression through a reduction in the number of polarized cells. Therefore, apical domain formation reinforces a TE fate. The results we present here demonstrate how polarization is triggered to regulate the first lineage segregation in human embryos.", "doi": "10.7554/eLife.65068", "issn": "2050-084X", "publisher": "eLife Sciences Publications", "publication": "eLife", "publication_date": "2021-09-27", "volume": "10", "pages": "Art. No. e65068" }, { "id": "authors:7pmdv-pjp54", "collection": "authors", "collection_id": "7pmdv-pjp54", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210317-080317393", "type": "article", "title": "Reconstructing aspects of human embryogenesis with pluripotent stem cells", "author": [ { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-Berna" }, { "family_name": "Jorgensen", "given_name": "Victoria", "orcid": "0000-0002-4205-6198", "clpid": "Jorgensen-Victoria" }, { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278", "clpid": "Weatherbee-Bailey-A-T" }, { "family_name": "Chen", "given_name": "Sisi", "orcid": "0000-0001-9448-9713", "clpid": "Chen-Sisi" }, { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Understanding human development is of fundamental biological and clinical importance. Despite its significance, mechanisms behind human embryogenesis remain largely unknown. Here, we attempt to model human early embryo development with expanded pluripotent stem cells (EPSCs) in 3-dimensions. We define a protocol that allows us to generate self-organizing cystic structures from human EPSCs that display some hallmarks of human early embryogenesis. These structures mimic polarization and cavitation characteristic of pre-implantation development leading to blastocyst morphology formation and the transition to post-implantation-like organization upon extended culture. Single-cell RNA sequencing of these structures reveals subsets of cells bearing some resemblance to epiblast, hypoblast and trophectoderm lineages. Nevertheless, significant divergences from natural blastocysts persist in some key markers, and signalling pathways point towards ways in which morphology and transcriptional-level cell identities may diverge in stem cell models of the embryo. Thus, this stem cell platform provides insights into the design of stem cell models of embryogenesis.", "doi": "10.1038/s41467-021-25853-4", "pmcid": "PMC8455697", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2021-09-21", "volume": "12", "pages": "Art. No. 5550" }, { "id": "authors:081bn-j0d09", "collection": "authors", "collection_id": "081bn-j0d09", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210930-211337700", "type": "article", "title": "An in vitro stem cell model of human epiblast and yolk sac interaction", "author": [ { "family_name": "Mackinlay", "given_name": "Kirsty M. L.", "orcid": "0000-0001-6276-4274", "clpid": "Mackinlay-Kirsty-M-L" }, { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278", "clpid": "Weatherbee-Bailey-A-T" }, { "family_name": "Souza Rosa", "given_name": "Viviane", "clpid": "Souza-Rosa-Viviane" }, { "family_name": "Handford", "given_name": "Charlotte E.", "clpid": "Handford-Charlotte-E" }, { "family_name": "Hudson", "given_name": "George", "orcid": "0000-0003-2506-6965", "clpid": "Hudson-George" }, { "family_name": "Coorens", "given_name": "Tim", "clpid": "Coorens-Tim" }, { "family_name": "Pereira", "given_name": "Lygia V.", "clpid": "Pereira-Lygia-V" }, { "family_name": "Behjati", "given_name": "Sam", "orcid": "0000-0002-6600-7665", "clpid": "Behjati-Sam" }, { "family_name": "Vallier", "given_name": "Ludovic", "orcid": "0000-0002-3848-2602", "clpid": "Vallier-Ludovic" }, { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-Marta-N" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Human embryogenesis entails complex signalling interactions between embryonic and extra-embryonic cells. However, how extra-embryonic cells direct morphogenesis within the human embryo remains largely unknown due to a lack of relevant stem cell models. Here, we have established conditions to differentiate human pluripotent stem cells (hPSCs) into yolk sac-like cells (YSLCs) that resemble the post-implantation human hypoblast molecularly and functionally. YSLCs induce the expression of pluripotency and anterior ectoderm markers in human embryonic stem cells (hESCs) at the expense of mesoderm and endoderm markers. This activity is mediated by the release of BMP and WNT signalling pathway inhibitors, and, therefore, resembles the functioning of the anterior visceral endoderm signalling centre of the mouse embryo, which establishes the anterior-posterior axis. Our results implicate the yolk sac in epiblast cell fate specification in the human embryo and propose YSLCs as a tool for studying post-implantation human embryo development in vitro.", "doi": "10.7554/elife.63930", "pmcid": "PMC8370770", "issn": "2050-084X", "publisher": "eLife Sciences Publications", "publication": "eLife", "publication_date": "2021-08-17", "volume": "10", "pages": "Art. No. e63930" }, { "id": "authors:wv5gj-gyb57", "collection": "authors", "collection_id": "wv5gj-gyb57", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210618-210040654", "type": "article", "title": "A single cell characterisation of human embryogenesis identifies pluripotency transitions and putative anterior hypoblast centre", "author": [ { "family_name": "Mol\u00e8", "given_name": "Matteo A.", "orcid": "0000-0001-7342-4849", "clpid": "Mol\u00e8-Matteo-A" }, { "family_name": "Coorens", "given_name": "Tim H. H.", "orcid": "0000-0002-5826-3554", "clpid": "Coorens-Tim-H-H" }, { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-Marta-N" }, { "family_name": "Weberling", "given_name": "Antonia", "orcid": "0000-0001-8282-5695", "clpid": "Weberling-Antonia" }, { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278", "clpid": "Weatherbee-Bailey-A-T" }, { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786", "clpid": "Gantner-Carlos-W" }, { "family_name": "Sancho-Serra", "given_name": "Carmen", "clpid": "Sancho-Serra-Carmen" }, { "family_name": "Richardson", "given_name": "Lucy", "clpid": "Richardson-Lucy" }, { "family_name": "Drinkwater", "given_name": "Abbie", "clpid": "Drinkwater-Abbie" }, { "family_name": "Syed", "given_name": "Najma", "clpid": "Syed-Najma" }, { "family_name": "Engley", "given_name": "Stephanie", "clpid": "Engley-Stephanie" }, { "family_name": "Snell", "given_name": "Philip", "clpid": "Snell-Philip" }, { "family_name": "Christie", "given_name": "Leila", "clpid": "Christie-Leila" }, { "family_name": "Elder", "given_name": "Kay", "orcid": "0000-0003-3510-8268", "clpid": "Elder-Kay" }, { "family_name": "Campbell", "given_name": "Alison", "clpid": "Campbell-Alison" }, { "family_name": "Fishel", "given_name": "Simon", "clpid": "Fishel-Simon" }, { "family_name": "Behjati", "given_name": "Sam", "orcid": "0000-0002-6600-7665", "clpid": "Behjati-Sam" }, { "family_name": "Vento-Tormo", "given_name": "Roser", "orcid": "0000-0002-9870-8474", "clpid": "Vento-Tormo-Roser" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Following implantation, the human embryo undergoes major morphogenetic transformations that establish the future body plan. While the molecular events underpinning this process are established in mice, they remain unknown in humans. Here we characterise key events of human embryo morphogenesis, in the period between implantation and gastrulation, using single-cell analyses and functional studies. First, the embryonic epiblast cells transition through different pluripotent states and act as a source of FGF signals that ensure proliferation of both embryonic and extra-embryonic tissues. In a subset of embryos, we identify a group of asymmetrically positioned extra-embryonic hypoblast cells expressing inhibitors of BMP, NODAL and WNT signalling pathways. We suggest that this group of cells can act as the anterior singalling centre to pattern the epiblast. These results provide insights into pluripotency state transitions, the role of FGF signalling and the specification of anterior-posterior axis during human embryo development.", "doi": "10.1038/s41467-021-23758-w", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2021-06-17", "volume": "12", "pages": "Art. No. 3679" }, { "id": "authors:qb3mc-07a51", "collection": "authors", "collection_id": "qb3mc-07a51", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210322-130659658", "type": "article", "title": "Unifying synthetic embryology", "author": [ { "family_name": "Cornwall-Scoones", "given_name": "Jake", "orcid": "0000-0002-7435-486X", "clpid": "Cornwall-Scoones-Jake" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "During embryonic development, a fertilized egg transitions into a complex organism, whereby diverse cell types are spatially organized into functional tissues. This sequential unfurling of complexity is exemplified perhaps best at gastrulation, where the major axes of the adult are laid down and the principal germ layers are defined in tandem with a dramatic change in morphology (Tam and Behringer, 1997; Arnold and Robertson, 2009). Understanding the mechanisms by which this highly choreographed process plays out demands investigation at different spatial scales (Shahbazi et al., 2019): from the cell-intrinsic gene-regulatory networks that govern cell fate decisions; to the signaling interactions that coordinate divergent fate trajectories across tissues. It requires understanding of how these regulatory processes are coupled to global characteristics including embryonic geometry, patterning, and inter-tissue interactions. Dissecting mechanisms across such scales \u2014 and crucially understanding their interactions \u2014 requires the capacity to manipulate genes, signals and morphology, a task that has proved challenging particularly for mammalian experimental embryology given the inaccessibility of the conceptus at implantation, a time when many of these important events occur (Hadjantonakis et al., 2020; Shahbazi and Zernicka-Goetz, 2018). Synthetic embryology has risen to this challenge, engineering embryo-like structures \u2014 \"stembryos\" \u2014 using stem cells derived from embryos situating cells in both near-native and more foreign contexts, and probing the consequences for patterning and morphogenesis. Given the recent explosion of diversity in stembryo models (Harrison et al., 2017; Rivron et al., 2018; Shao et al., 2017a; Sozen et al., 2018; Van den Brink et al., 2014; Veenvliet et al., 2020; Warmflash et al., 2014; Amadei et al., 2020), this Special Issue reflects on the successes made and the challenges that remain in using in vitro culture platforms to study early mammalian embryogenesis. Here, we offer a perspective on the purpose and utility of synthetic embryology, proposing that focusing efforts on probing fundamental mechanisms of self-organization, spurred via novel bioengineering strategies, can help unify the field towards its shared motivations of understanding natural development and building reliable, translatable experimental models.", "doi": "10.1016/j.ydbio.2021.03.007", "pmcid": "PMC8257322", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2021-06", "volume": "474", "pages": "1-4" }, { "id": "authors:5z1b8-hjd91", "collection": "authors", "collection_id": "5z1b8-hjd91", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201217-133744809", "type": "article", "title": "The dynamics of morphogenesis in stem cell-based embryology: Novel insights for symmetry breaking", "author": [ { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-Berna" }, { "family_name": "Cornwall-Scoones", "given_name": "Jake", "orcid": "0000-0002-7435-486X", "clpid": "Cornwall-Scoones-Jake" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Breaking embryonic symmetry is an essential prerequisite to shape the initially symmetric embryo into a highly organized body plan that serves as the blueprint of the adult organism. This critical process is driven by morphogen signaling gradients that instruct anteroposterior axis specification. Despite its fundamental importance, what triggers symmetry breaking and how the signaling gradients are established in time and space in the mammalian embryo remain largely unknown. Stem cell-based in vitro models of embryogenesis offer an unprecedented opportunity to quantitatively dissect the multiple physical and molecular processes that shape the mammalian embryo. Here we review biochemical mechanisms governing early mammalian patterning in vivo and highlight recent advances to recreate this in vitro using stem cells. We discuss how the novel insights from these model systems extend previously proposed concepts to illuminate the extent to which embryonic cells have the intrinsic capability to generate specific, reproducible patterns during embryogenesis.", "doi": "10.1016/j.ydbio.2020.12.005", "pmcid": "PMC8259461", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2021-06", "volume": "474", "pages": "82-90" }, { "id": "authors:nmyqc-63778", "collection": "authors", "collection_id": "nmyqc-63778", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201217-133744400", "type": "article", "title": "Modeling human embryo development with embryonic and extra-embryonic stem cells", "author": [ { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278", "clpid": "Weatherbee-Bailey-A-T" }, { "family_name": "Cui", "given_name": "Tongtong", "orcid": "0000-0002-2474-3832", "clpid": "Cui-Tongtong" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Early human post-implantation development involves extensive growth combined with a series of complex morphogenetic events. The lack of precise spatial and temporal control over these processes leads to pregnancy loss. Given the ethical and technical limitations in studying the natural human embryo, alternative approaches are needed to investigate mechanisms underlying this critical stage of human development. Here, we present an overview of the different stem cells and stem cell-derived models which serve as useful, albeit imperfect, tools in understanding human embryogenesis. Current models include stem cells that represent each of the three earliest lineages: human embryonic stem cells corresponding to the epiblast, hypoblast-like stem cells and trophoblast stem cells. We also review the use of human embryonic stem cells to model complex aspects of epiblast morphogenesis and differentiation. Additionally, we propose that the combination of both embryonic and extra-embryonic stem cells to form three-dimensional embryo models will provide valuable insights into cell-cell chemical and mechanical interactions that are essential for natural embryogenesis.", "doi": "10.1016/j.ydbio.2020.12.010", "pmcid": "PMC8232073", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2021-06", "volume": "474", "pages": "91-99" }, { "id": "authors:y61cc-1kg62", "collection": "authors", "collection_id": "y61cc-1kg62", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210504-094816688", "type": "article", "title": "Machine learning-assisted high-content analysis of pluripotent stem cell-derived embryos in\u00a0vitro", "author": [ { "family_name": "Guo", "given_name": "Jianying", "clpid": "Guo-Jianying" }, { "family_name": "Wang", "given_name": "Peizhe", "clpid": "Wang-Peizhe" }, { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-Berna" }, { "family_name": "Qiu", "given_name": "Hui", "clpid": "Qiu-Hui" }, { "family_name": "Zhu", "given_name": "Yonglin", "clpid": "Zhu-Yonglin" }, { "family_name": "Zhang", "given_name": "Xingwu", "clpid": "Zhang-Xingwu" }, { "family_name": "Ming", "given_name": "Jia", "clpid": "Ming-Jia" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Na", "given_name": "Jie", "orcid": "0000-0003-1820-0548", "clpid": "Na-Jie" } ], "abstract": "Stem cell-based embryo models by cultured pluripotent and extra-embryonic lineage stem cells are novel platforms to model early postimplantation development. We showed that induced pluripotent stem cells (iPSCs) could form ITS (iPSCs and trophectoderm stem cells) and ITX (iPSCs, trophectoderm stem cells, and XEN cells) embryos, resembling the early gastrula embryo developed in vivo. To facilitate the efficient and unbiased analysis of the stem cell-based embryo model, we set up a machine learning workflow to extract multi-dimensional features and perform quantification of ITS embryos using 3D images collected from a high-content screening system. We found that different PSC lines differ in their ability to form embryo-like structures. Through high-content screening of small molecules and cytokines, we identified that BMP4 best promoted the morphogenesis of the ITS embryo. Our study established an innovative strategy to analyze stem cell-based embryo models and uncovered new roles of BMP4 in stem cell-based embryo models.", "doi": "10.1016/j.stemcr.2021.03.018", "pmcid": "PMC8185434", "issn": "2213-6711", "publisher": "Cell Press", "publication": "Stem Cell Reports", "publication_date": "2021-05-11", "series_number": "5", "volume": "16", "issue": "5", "pages": "1331-1346" }, { "id": "authors:f1y3m-sn581", "collection": "authors", "collection_id": "f1y3m-sn581", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201009-083350141", "type": "article", "title": "Embryo Size Regulates the Timing and Mechanism of Pluripotent Tissue Morphogenesis", "author": [ { "family_name": "Orietti", "given_name": "Lorenzo C.", "clpid": "Orietti-Lorenzo-C" }, { "family_name": "Rosa", "given_name": "Viviane Souza", "clpid": "Rosa-Viviane-Souza" }, { "family_name": "Antonica", "given_name": "Francesco", "orcid": "0000-0001-9359-9689", "clpid": "Antonica-Francesco" }, { "family_name": "Kyprianou", "given_name": "Christos", "orcid": "0000-0001-8516-9195", "clpid": "Kyprianou-Christos" }, { "family_name": "Mansfield", "given_name": "William", "clpid": "Mansfield-William" }, { "family_name": "Marques-Souza", "given_name": "Henrique", "orcid": "0000-0002-5008-8413", "clpid": "Marques-Souza-Henrique" }, { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-Marta-N" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Mammalian embryogenesis is a paradigm of regulative development as mouse embryos show plasticity in the regulation of cell fate, cell number, and tissue morphogenesis. However, the mechanisms behind embryo plasticity remain largely unknown. Here, we determine how mouse embryos respond to an increase in cell numbers to regulate the timing and mechanism of embryonic morphogenesis, leading to the formation of the pro-amniotic cavity. Using embryos and embryonic stem cell aggregates of different size, we show that while pro-amniotic cavity formation in normal-sized embryos is achieved through basement membrane-induced polarization and exocytosis, cavity formation of increased-size embryos is delayed and achieved through apoptosis of cells that lack contact with the basement membrane. Importantly, blocking apoptosis, both genetically and pharmacologically, alters pro-amniotic cavity formation but does not affect size regulation in enlarged embryos. We conclude that the regulation of embryonic size and morphogenesis, albeit concomitant, have distinct molecular underpinnings.", "doi": "10.1016/j.stemcr.2020.09.004", "pmcid": "PMC8185375", "issn": "2213-6711", "publisher": "Cell Press", "publication": "Stem Cell Reports", "publication_date": "2021-05-11", "series_number": "5", "volume": "16", "issue": "5", "pages": "1182-1196" }, { "id": "authors:y1arb-67246", "collection": "authors", "collection_id": "y1arb-67246", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210211-165540246", "type": "article", "title": "PANDORA-seq expands the repertoire of regulatory small RNAs by overcoming RNA modifications", "author": [ { "family_name": "Shi", "given_name": "Junchao", "orcid": "0000-0002-1548-5069", "clpid": "Shi-Junchao" }, { "family_name": "Zhang", "given_name": "Yunfang" }, { "family_name": "Tan", "given_name": "Dongmei" }, { "family_name": "Zhang", "given_name": "Xudong" }, { "family_name": "Yan", "given_name": "Menghong" }, { "family_name": "Zhang", "given_name": "Ying" }, { "family_name": "Franklin", "given_name": "Reuben" }, { "family_name": "Shahbazi", "given_name": "Marta", "orcid": "0000-0002-1599-5747" }, { "family_name": "Mackinlay", "given_name": "Kirsty" }, { "family_name": "Liu", "given_name": "Shichao" }, { "family_name": "Kuhle", "given_name": "Bernhard", "orcid": "0000-0002-0501-1150" }, { "family_name": "James", "given_name": "Emma R." }, { "family_name": "Zhang", "given_name": "Liwen" }, { "family_name": "Qu", "given_name": "Yongcun" }, { "family_name": "Zhai", "given_name": "Qiwei", "orcid": "0000-0002-9617-7403" }, { "family_name": "Zhao", "given_name": "Wenxin" }, { "family_name": "Zhao", "given_name": "Linlin", "orcid": "0000-0002-8821-4198" }, { "family_name": "Zhou", "given_name": "Changcheng", "orcid": "0000-0001-7649-0710" }, { "family_name": "Gu", "given_name": "Weifeng" }, { "family_name": "Murn", "given_name": "Jernej" }, { "family_name": "Guo", "given_name": "Jingtao" }, { "family_name": "Carrell", "given_name": "Douglas T.", "orcid": "0000-0002-6471-2803" }, { "family_name": "Wang", "given_name": "Yinsheng", "orcid": "0000-0001-5565-283X" }, { "family_name": "Chen", "given_name": "Xuemei", "orcid": "0000-0002-5209-1157" }, { "family_name": "Cairns", "given_name": "Bradley R.", "orcid": "0000-0002-9864-8811" }, { "family_name": "Yang", "given_name": "Xiang-lei", "orcid": "0000-0003-2554-084X" }, { "family_name": "Schimmel", "given_name": "Paul", "orcid": "0000-0002-0448-022X" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Cheloufi", "given_name": "Sihem", "orcid": "0000-0002-1726-4796" }, { "family_name": "Zhang", "given_name": "Ying", "clpid": "Zhang-Ying" }, { "family_name": "Zhou", "given_name": "Tong", "orcid": "0000-0003-2361-1931" }, { "family_name": "Chen", "given_name": "Qi", "orcid": "0000-0001-6353-9589" } ], "abstract": "Although high-throughput RNA sequencing (RNA-seq) has greatly advanced small non-coding RNA (sncRNA) discovery, the currently widely used complementary DNA library construction protocol generates biased sequencing results. This is partially due to RNA modifications that interfere with adapter ligation and reverse transcription processes, which prevent the detection of sncRNAs bearing these modifications. Here, we present PANDORA-seq (panoramic RNA display by overcoming RNA modification aborted sequencing), employing a combinatorial enzymatic treatment to remove key RNA modifications that block adapter ligation and reverse transcription. PANDORA-seq identified abundant modified sncRNAs\u2014mostly transfer RNA-derived small RNAs (tsRNAs) and ribosomal RNA-derived small RNAs (rsRNAs)\u2014that were previously undetected, exhibiting tissue-specific expression across mouse brain, liver, spleen and sperm, as well as cell-specific expression across embryonic stem cells (ESCs) and HeLa cells. Using PANDORA-seq, we revealed unprecedented landscapes of microRNA, tsRNA and rsRNA dynamics during the generation of induced pluripotent stem cells. Importantly, tsRNAs and rsRNAs that are downregulated during somatic cell reprogramming impact cellular translation in ESCs, suggesting a role in lineage differentiation.", "doi": "10.1038/s41556-021-00652-7", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2021-04", "series_number": "4", "volume": "23", "issue": "4", "pages": "424-436" }, { "id": "authors:98k8h-qfv94", "collection": "authors", "collection_id": "98k8h-qfv94", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210310-141520229", "type": "article", "title": "Integrin \u03b21 coordinates survival and morphogenesis of the embryonic lineage upon implantation and pluripotency transition", "author": [ { "family_name": "Mol\u00e8", "given_name": "Matteo Amitaba", "orcid": "0000-0001-7342-4849", "clpid": "Mol\u00e8-Matteo-Amitaba" }, { "family_name": "Weberling", "given_name": "Antonia", "orcid": "0000-0001-8282-5695", "clpid": "Weberling-Antonia" }, { "family_name": "F\u00e4ssler", "given_name": "Reinhard", "clpid": "F\u00e4ssler-Reinhard" }, { "family_name": "Campbell", "given_name": "Alison", "orcid": "0000-0002-4065-5672", "clpid": "Campbell-Alison" }, { "family_name": "Fishel", "given_name": "Simon", "clpid": "Fishel-Simon" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "At implantation, the embryo establishes contacts with the maternal endometrium. This stage is associated with a high incidence of preclinical pregnancy losses. While the maternal factors underlying uterine receptivity have been investigated, the signals required by the embryo for successful peri-implantation development remain elusive. To explore these, we studied integrin \u03b21 signaling, as embryos deficient for this receptor degenerate at implantation. We demonstrate that the coordinated action of pro-survival signals and localized actomyosin suppression via integrin \u03b21 permits the development of the embryo beyond implantation. Failure of either process leads to developmental arrest and apoptosis. Pharmacological stimulation through fibroblast growth factor 2 (FGF2) and insulin-like growth factor 1 (IGF1), coupled with ROCK-mediated actomyosin inhibition, rescues the deficiency of integrin \u03b21, promoting progression to post-implantation stages. Mutual exclusion between integrin \u03b21 and actomyosin seems to be conserved in the human embryo, suggesting the possibility that these mechanisms could also underlie the transition of the human epiblast from pre- to post-implantation.", "doi": "10.1016/j.celrep.2021.108834", "pmcid": "PMC7966855", "issn": "2211-1247", "publisher": "Cell Press", "publication": "Cell Reports", "publication_date": "2021-03-09", "series_number": "10", "volume": "34", "issue": "10", "pages": "Art. No. 108834" }, { "id": "authors:zkpyq-38e39", "collection": "authors", "collection_id": "zkpyq-38e39", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210104-164232285", "type": "article", "title": "Inducible Stem-Cell-Derived Embryos Capture Mouse Morphogenetic Events In\u00a0Vitro", "author": [ { "family_name": "Amadei", "given_name": "Gianluca", "orcid": "0000-0001-5405-968X", "clpid": "Amadei-Gianluca" }, { "family_name": "Lau", "given_name": "Kasey Y. C.", "clpid": "Lau-Kasey-Y-C" }, { "family_name": "De Jonghe", "given_name": "Joachim", "orcid": "0000-0003-0584-8265", "clpid": "De-Jonghe-Joachim" }, { "family_name": "Gantner", "given_name": "Carlos W.", "orcid": "0000-0003-0825-7786", "clpid": "Gantner-Carlos-W" }, { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-Berna" }, { "family_name": "Chan", "given_name": "Christopher", "clpid": "Chan-Christopher" }, { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Kyprianou", "given_name": "Christos", "orcid": "0000-0001-8516-9195", "clpid": "Kyprianou-Christos" }, { "family_name": "Hollfelder", "given_name": "Florian", "orcid": "0000-0002-1367-6312", "clpid": "Hollfelder-Florian" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The development of mouse embryos can be partially recapitulated by combining embryonic stem cells (ESCs), trophoblast stem cells (TS), and extra-embryonic endoderm (XEN) stem cells to generate embryo-like structures called ETX embryos. Although ETX embryos transcriptionally capture the mouse gastrula, their ability to recapitulate complex morphogenic events such as gastrulation is limited, possibly due to the limited potential of XEN cells. To address this, we generated ESCs transiently expressing transcription factor Gata4, which drives the extra-embryonic endoderm fate, and combined them with ESCs and TS cells to generate induced ETX embryos (iETX embryos). We show that iETX embryos establish a robust anterior signaling center that migrates unilaterally to break embryo symmetry. Furthermore, iETX embryos gastrulate generating embryonic and extra-embryonic mesoderm and definitive endoderm. Our findings reveal that replacement of XEN cells with ESCs transiently expressing Gata4 endows iETX embryos with greater developmental potential, thus enabling the study of the establishment of anterior-posterior patterning and gastrulation in an in vitro system.", "doi": "10.1016/j.devcel.2020.12.004", "pmcid": "PMC7883308", "issn": "1534-5807", "publisher": "Cell Press", "publication": "Developmental Cell", "publication_date": "2021-02-08", "series_number": "3", "volume": "56", "issue": "3", "pages": "366-382" }, { "id": "authors:hm711-1m846", "collection": "authors", "collection_id": "hm711-1m846", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201117-110225184", "type": "article", "title": "BMP signalling is required for extra-embryonic ectoderm development during pre-to-post-implantation transition of the mouse embryo", "author": [ { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-Berna" }, { "family_name": "Demir", "given_name": "Necdet", "orcid": "0000-0002-8166-5535", "clpid": "Demir-Necdet" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "At implantation, the mouse embryo undergoes a critical transformation which requires the precise spatiotemporal control of signalling pathways necessary for morphogenesis and developmental progression. The role played by such signalling pathways during this transition are largely unexplored, due to the inaccessibility of the embryo during the implantation when it becomes engulfed by uterine tissues. Genetic studies demonstrate that mutant embryos for BMPs die around gastrulation. Here we have aimed to dissect the role of BMPs during pre-to post-implantation transition by using a protocol permitting the development of the embryo beyond implantation stages in vitro and using stem cells to mimic post-implantation tissue organisation. By assessing both the canonical and non-canonical mechanisms of BMP, we show that the loss of canonical BMP activity compromises the extra-embryonic ectoderm development. Our analyses demonstrate that BMP signalling maintains stem cell populations within both embryonic/extra-embryonic tissues during pre-to post-implantation development. These results may provide insight into the role played by BMP signalling in controlling early embryogenesis.", "doi": "10.1016/j.ydbio.2020.11.005", "pmcid": "PMC8219371", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2021-02", "volume": "470", "pages": "84-94" }, { "id": "authors:nr1ny-74q16", "collection": "authors", "collection_id": "nr1ny-74q16", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20210119-143307098", "type": "article", "title": "Trophectoderm mechanics direct epiblast shape upon embryo implantation", "author": [ { "family_name": "Weberling", "given_name": "Antonia", "orcid": "0000-0001-8282-5695", "clpid": "Weberling-Antonia" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Implantation is a hallmark of mammalian embryogenesis during which embryos establish their contacts with the maternal endometrium, remodel, and undertake growth and differentiation. The mechanisms and sequence of events through which embryos change their shape during this transition are largely unexplored. Here, we show that the first extraembryonic lineage, the polar trophectoderm, is the key regulator for remodeling the embryonic epiblast. Loss of its function after immuno-surgery or inhibitor treatments prevents the epiblast shape transitions. In the mouse, the polar trophectoderm exerts physical force upon the epiblast, causing it to transform from an oval into a cup shape. In human embryos, the polar trophectoderm behaves in the opposite manner, exerting a stretching force. By mimicking this stretching behavior in mouse embryogenesis, we could direct the epiblast to adopt the disc-like shape characteristic of human embryos at this stage. Thus, the polar trophectoderm acts as a conserved regulator of epiblast shape.", "doi": "10.1016/j.celrep.2020.108655", "pmcid": "PMC7816124", "issn": "2211-1247", "publisher": "Cell Press", "publication": "Cell Reports", "publication_date": "2021-01-19", "series_number": "3", "volume": "34", "issue": "3", "pages": "Art. No. 108655" }, { "id": "authors:teegz-hk924", "collection": "authors", "collection_id": "teegz-hk924", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200212-084819179", "type": "article", "title": "Developmental clock and mechanism of de novo polarization of the mouse embryo", "author": [ { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Cornwall-Scoones", "given_name": "Jake", "orcid": "0000-0002-7435-486X", "clpid": "Cornwall-Scoones-Jake" }, { "family_name": "Wang", "given_name": "Peizhe", "clpid": "Wang-Peizhe" }, { "family_name": "Handford", "given_name": "Charlotte E.", "orcid": "0000-0002-5245-8027", "clpid": "Handford-Charlotte-E" }, { "family_name": "Na", "given_name": "Jie", "orcid": "0000-0003-1820-0548", "clpid": "Na-Jie" }, { "family_name": "Thomson", "given_name": "Matt", "orcid": "0000-0003-1021-1234", "clpid": "Thomson-M-W" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Embryo polarization is critical for mouse development; however, neither the regulatory clock nor the molecular trigger that it activates is known. Here, we show that the embryo polarization clock reflects the onset of zygotic genome activation, and we identify three factors required to trigger polarization. Advancing the timing of transcription factor AP-2 gamma (Tfap2c) and TEA domain transcription factor 4 (Tead4) expression in the presence of activated Ras homolog family member A (RhoA) induces precocious polarization as well as subsequent cell fate specification and morphogenesis. Tfap2c and Tead4 induce expression of actin regulators that control the recruitment of apical proteins on the membrane, whereas RhoA regulates their lateral mobility, allowing the emergence of the apical domain. Thus, Tfap2c, Tead4, and RhoA are regulators for the onset of polarization and cell fate segregation in the mouse.", "doi": "10.1126/science.abd2703", "pmcid": "PMC8210885", "issn": "0036-8075", "publisher": "American Association for the Advancement of Science", "publication": "Science", "publication_date": "2020-12-11", "series_number": "6522", "volume": "370", "issue": "6522", "pages": "Art. No. eabd2703" }, { "id": "authors:yf3k4-4mz35", "collection": "authors", "collection_id": "yf3k4-4mz35", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201210-160239379", "type": "article", "title": "Principles of Self-Organization of the Mammalian Embryo", "author": [ { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Early embryogenesis is a conserved and self-organized process. In the mammalian embryo, the potential for self-organization is manifested in its extraordinary developmental plasticity, allowing a correctly patterned embryo to arise despite experimental perturbation. The underlying mechanisms enabling such regulative development have long been a topic of study. In this Review, we summarize our current understanding of the self-organizing principles behind the regulative nature of the early mammalian embryo. We argue that geometrical constraints, feedback between mechanical and biochemical factors, and cellular heterogeneity are all required to ensure the developmental plasticity of mammalian embryo development.", "doi": "10.1016/j.cell.2020.11.003", "pmcid": "PMC8212876", "issn": "0092-8674", "publisher": "Cell Press", "publication": "Cell", "publication_date": "2020-12-10", "series_number": "6", "volume": "183", "issue": "6", "pages": "1467-1478" }, { "id": "authors:yez9s-st641", "collection": "authors", "collection_id": "yez9s-st641", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201012-163632801", "type": "article", "title": "Global hyperactivation of enhancers stabilizes human and mouse naive pluripotency through inhibition of CDK8/19 Mediator kinases", "author": [ { "family_name": "Lynch", "given_name": "Cian J.", "clpid": "Lynch-Cian-J" }, { "family_name": "Bernad", "given_name": "Raquel" }, { "family_name": "Mart\u00ednez-Val", "given_name": "Ana" }, { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747" }, { "family_name": "N\u00f3brega-Pereira", "given_name": "Sandrina", "orcid": "0000-0002-9652-1382" }, { "family_name": "Calvo", "given_name": "Isabel", "orcid": "0000-0003-4107-296X" }, { "family_name": "Blanco-Aparicio", "given_name": "Carmen", "orcid": "0000-0002-3249-6595" }, { "family_name": "Tarantino", "given_name": "Carolina" }, { "family_name": "Garreta", "given_name": "Elena" }, { "family_name": "Richart-Gin\u00e9s", "given_name": "Laia" }, { "family_name": "Alcazar", "given_name": "Noelia" }, { "family_name": "Gra\u00f1a-Castro", "given_name": "Osvaldo" }, { "family_name": "G\u00f3mez-Lopez", "given_name": "Gonzalo", "orcid": "0000-0002-4146-0551" }, { "family_name": "Aksoy", "given_name": "Irene" }, { "family_name": "Mu\u00f1oz-Mart\u00edn", "given_name": "Maribel" }, { "family_name": "Martinez", "given_name": "Sonia", "orcid": "0000-0003-2230-7794" }, { "family_name": "Ortega", "given_name": "Sagrario" }, { "family_name": "Prieto", "given_name": "Susana", "orcid": "0000-0002-9746-2396" }, { "family_name": "Simboeck", "given_name": "Elisabeth" }, { "family_name": "Camasses", "given_name": "Alain" }, { "family_name": "Stephan-Otto Attolini", "given_name": "Camille", "orcid": "0000-0001-8045-320X" }, { "family_name": "Fernandez", "given_name": "Agustin F.", "orcid": "0000-0002-3792-4085" }, { "family_name": "Sierra", "given_name": "Marta I." }, { "family_name": "Fraga", "given_name": "Mario F." }, { "family_name": "Pastor", "given_name": "Joaquin" }, { "family_name": "Fisher", "given_name": "Daniel", "orcid": "0000-0002-0822-3482" }, { "family_name": "Montserrat", "given_name": "Nuria" }, { "family_name": "Savatier", "given_name": "Pierre" }, { "family_name": "Mu\u00f1oz", "given_name": "Javier", "orcid": "0000-0003-3288-3496" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Serrano", "given_name": "Manuel", "orcid": "0000-0001-7177-9312" } ], "abstract": "Pluripotent stem cells (PSCs) transition between cell states in vitro, reflecting developmental changes in the early embryo. PSCs can be stabilized in the naive state by blocking extracellular differentiation stimuli, particularly FGF\u2013MEK signalling. Here, we report that multiple features of the naive state in human and mouse PSCs can be recapitulated without affecting FGF\u2013MEK signalling or global DNA methylation. Mechanistically, chemical inhibition of CDK8 and CDK19 (hereafter CDK8/19) kinases removes their ability to repress the Mediator complex at enhancers. CDK8/19 inhibition therefore increases Mediator-driven recruitment of RNA polymerase II (RNA Pol II) to promoters and enhancers. This efficiently stabilizes the naive transcriptional program and confers resistance to enhancer perturbation by BRD4 inhibition. Moreover, naive pluripotency during embryonic development coincides with a reduction in CDK8/19. We conclude that global hyperactivation of enhancers drives naive pluripotency, and this can be achieved in vitro by inhibiting CDK8/19 kinase activity. These principles may apply to other contexts of cellular plasticity.", "doi": "10.1038/s41556-020-0573-1", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2020-10", "series_number": "10", "volume": "22", "issue": "10", "pages": "1223-1238" }, { "id": "authors:af06e-z0486", "collection": "authors", "collection_id": "af06e-z0486", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20201009-152128912", "type": "article", "title": "Starting life in space", "author": [ { "family_name": "Cornwall-Scoones", "given_name": "Jake", "clpid": "Cornwall-Scoones-J" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "As far as we know, animal development is a process that is unique to our planet. That does not mean, however, that development beyond our realm is impossible. As it starts to become feasible for us to look to the sky for another place to call home, we may start to appreciate the gravity of this question. \n\nIn this issue, Lei et al. investigate for the first time the repercussions of space travel on the first decisions made by mouse embryos. Developing a novel micro-incubator, capable of automatic micrography and fixation, harboring some 3400 two-cell embryos, they investigate the consequences of development after being projected into the stratosphere in the SJ-10 satellite [1].", "doi": "10.1093/nsr/nwaa102", "issn": "2095-5138", "publisher": "Oxford University Press", "publication": "National Science Review", "publication_date": "2020-09-01", "series_number": "9", "volume": "7", "issue": "9", "pages": "1447-1448" }, { "id": "authors:yg1q4-rj583", "collection": "authors", "collection_id": "yg1q4-rj583", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200811-114715166", "type": "article", "title": "Developmental potential of aneuploid human embryos cultured beyond implantation", "author": [ { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-M-N" }, { "family_name": "Wang", "given_name": "Tianren", "clpid": "Wang-Tianren" }, { "family_name": "Tao", "given_name": "Xin", "clpid": "Tao-Xin" }, { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "orcid": "0000-0002-6825-6278", "clpid": "Weatherbee-B-A-T" }, { "family_name": "Sun", "given_name": "Li", "clpid": "Sun-Li" }, { "family_name": "Zhan", "given_name": "Yiping", "clpid": "Zhan-Yiping" }, { "family_name": "Keller", "given_name": "Laura", "clpid": "Keller-Laura" }, { "family_name": "Smith", "given_name": "Gary D.", "clpid": "Smith-Gary-D" }, { "family_name": "Pellicer", "given_name": "Antonio", "orcid": "0000-0002-8254-863X", "clpid": "Pellicer-A" }, { "family_name": "Scott", "given_name": "Richard T., Jr.", "clpid": "Scott-R-T-Jr" }, { "family_name": "Seli", "given_name": "Emre", "orcid": "0000-0001-7464-8203", "clpid": "Seli-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Aneuploidy, the presence of an abnormal number of chromosomes, is a major cause of early pregnancy loss in humans. Yet, the developmental consequences of specific aneuploidies remain unexplored. Here, we determine the extent of post-implantation development of human embryos bearing common aneuploidies using a recently established culture platform. We show that while trisomy 15 and trisomy 21 embryos develop similarly to euploid embryos, monosomy 21 embryos exhibit high rates of developmental arrest, and trisomy 16 embryos display a hypo-proliferation of the trophoblast, the tissue that forms the placenta. Using human trophoblast stem cells, we show that this phenotype can be mechanistically ascribed to increased levels of the cell adhesion protein E-CADHERIN, which lead to premature differentiation and cell cycle arrest. We identify three cases of mosaicism in embryos diagnosed as full aneuploid by pre-implantation genetic testing. Our results present the first detailed analysis of post-implantation development of aneuploid human embryos.", "doi": "10.1038/s41467-020-17764-7", "pmcid": "PMC7418029", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2020-08-10", "volume": "11", "pages": "Art. No. 3987" }, { "id": "authors:y1qv7-23k59", "collection": "authors", "collection_id": "y1qv7-23k59", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200805-090719433", "type": "article", "title": "Expression of SARS-CoV-2 receptor ACE2 and the protease TMPRSS2 suggests susceptibility of the human embryo in the first trimester", "author": [ { "family_name": "Weatherbee", "given_name": "Bailey A. T.", "clpid": "Weatherbee-B-A-T" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "While initially recognized as causing respiratory disease, the SARS-CoV-2 virus also affects many other organs leading to other complications. It has emerged that advanced age and obesity are risk factors for complications but questions concerning the potential effects on fetal health and successful pregnancy for those infected with SARS-CoV-2 remain largely unanswered. Here, we examine human pre-gastrulation embryos to determine the expression patterns of the genes ACE2, encoding the SARS-CoV-2 receptor, and TMPRSS2, encoding a protease that cleaves both the viral spike protein and the ACE2 receptor to facilitate infection. We show expression and co-expression of these genes in the trophoblast of the blastocyst and syncytiotrophoblast and hypoblast of the implantation stages, which develop into tissues that interact with the maternal blood supply for nutrient exchange. Expression of ACE2 and TMPRSS2 in these tissues raises the possibility for vertical transmission and indicates that further work is required to understand potential risks to implantation, placental health and fetal health that require further study.", "doi": "10.1098/rsob.200162", "pmcid": "PMC7479935", "issn": "2046-2441", "publisher": "Royal Society", "publication": "Open Biology", "publication_date": "2020-08", "series_number": "8", "volume": "10", "issue": "8", "pages": "200162" }, { "id": "authors:kk6qe-2mh31", "collection": "authors", "collection_id": "kk6qe-2mh31", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200611-161658303", "type": "article", "title": "Autophagy-mediated apoptosis eliminates aneuploid cells in a mouse model of chromosome mosaicism", "author": [ { "family_name": "Singla", "given_name": "Shruti", "clpid": "Singla-Shruti" }, { "family_name": "Iwamoto-Stohl", "given_name": "Lisa K.", "clpid": "Iwamoto-Stohl-L-K" }, { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The high incidence of aneuploidy in the embryo is considered the principal cause for low human fecundity. However, the prevalence of aneuploidy dramatically declines as pregnancy progresses, with the steepest drop occurring as the embryo completes implantation. Despite the fact that the plasticity of the embryo in dealing with aneuploidy is fundamental to normal development, the mechanisms responsible for eliminating aneuploid cells are unclear. Here, using a mouse model of chromosome mosaicism, we show that aneuploid cells are preferentially eliminated from the embryonic lineage in a p53-dependent process involving both autophagy and apoptosis before, during and after implantation. Moreover, we show that diploid cells in mosaic embryos undertake compensatory proliferation during the implantation stages to confer embryonic viability. Together, our results indicate a close link between aneuploidy, autophagy, and apoptosis to refine the embryonic cell population and ensure only chromosomally fit cells proceed through development of the fetus.", "doi": "10.1038/s41467-020-16796-3", "pmcid": "PMC7290028", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2020-06-11", "volume": "11", "pages": "Art. No. 2958" }, { "id": "authors:mrkpc-7a922", "collection": "authors", "collection_id": "mrkpc-7a922", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200121-134023730", "type": "article", "title": "Basement membrane remodelling regulates mouse embryogenesis", "author": [ { "family_name": "Kyprianou", "given_name": "Christos", "orcid": "0000-0001-8516-9195", "clpid": "Kyprianou-Christos" }, { "family_name": "Christodoulou", "given_name": "Neophytos", "orcid": "0000-0003-4338-5847", "clpid": "Christodoulou-Neophytos" }, { "family_name": "Hamilton", "given_name": "Russell S.", "clpid": "Hamilton-Russell-S" }, { "family_name": "Nahaboo", "given_name": "Wallis", "clpid": "Nahaboo-Wallis" }, { "family_name": "Boomgaard", "given_name": "Diana Suarez", "clpid": "Boomgaard-Diana-Suarez" }, { "family_name": "Amadei", "given_name": "Gianluca", "clpid": "Amadei-Gianluca" }, { "family_name": "Migeotte", "given_name": "Isabelle", "clpid": "Migeotte-Isabelle" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Tissue sculpting during development has been attributed mainly to cellular events through processes such as convergent extension or apical constriction. However, recent work has revealed roles for basement membrane remodelling in global tissue morphogenesis. Upon implantation, the epiblast and extraembryonic ectoderm of the mouse embryo become enveloped by a basement membrane. Signalling between the basement membrane and these tissues is critical for cell polarization and the ensuing morphogenesis. However, the mechanical role of the basement membrane in post-implantation embryogenesis remains unknown. Here we demonstrate the importance of spatiotemporally regulated basement membrane remodelling during early embryonic development. Specifically, we show that Nodal signalling directs the generation and dynamic distribution of perforations in the basement membrane by regulating the expression of matrix metalloproteinases. This basement membrane remodelling facilitates embryo growth before gastrulation. The establishment of the anterior\u2013posterior axis further regulates basement membrane remodelling by localizing Nodal signalling\u2014and therefore the activity of matrix metalloproteinases and basement membrane perforations\u2014to the posterior side of the embryo. Perforations on the posterior side are essential for primitive-streak extension during gastrulation by rendering the basement membrane of the prospective primitive streak more prone to breaching. Thus spatiotemporally regulated basement membrane remodelling contributes to the coordination of embryo growth, morphogenesis and gastrulation.", "doi": "10.1038/s41586-020-2264-2", "pmcid": "PMC7308173", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2020-06-11", "series_number": "7811", "volume": "582", "issue": "7811", "pages": "253-258" }, { "id": "authors:hyg55-z7g80", "collection": "authors", "collection_id": "hyg55-z7g80", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200406-124658130", "type": "article", "title": "Living a Sweet Life: Glucose Instructs Cell Fate in the Mouse Embryo", "author": [ { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "It has long been known that glucose is required for the development of pre-implantation mouse embryos, but the mechanism accounting for such a requirement has remained unknown. In this issue of Developmental Cell, Chi et al. dissect the molecular pathways that respond to the state of glucose metabolism to drive the morula to blastocyst transition.", "doi": "10.1016/j.devcel.2020.03.012", "issn": "1534-5807", "publisher": "Cell Press", "publication": "Developmental Cell", "publication_date": "2020-04-06", "series_number": "1", "volume": "53", "issue": "1", "pages": "1-2" }, { "id": "authors:esqhn-cgd91", "collection": "authors", "collection_id": "esqhn-cgd91", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200409-115737960", "type": "article", "title": "Building an apical domain in the early mouse embryo: Lessons, challenges and perspectives", "author": [ { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Cell polarization is critical for lineage segregation and morphogenesis during mammalian embryogenesis. However, the processes and mechanisms that establish cell polarity in the mammalian embryo are not well understood. Recent studies suggest that unique regulatory mechanisms are deployed by the mouse embryo to establish cell polarization. In this review, we discuss the current understanding of cell polarity establishment, focusing on the formation of the apical domain in the mouse embryo. We will also discuss outstanding questions and possible directions for future study.", "doi": "10.1016/j.ceb.2019.11.005", "issn": "0955-0674", "publisher": "Elsevier", "publication": "Current Opinion in Cell Biology", "publication_date": "2020-02", "volume": "62", "pages": "144-149" }, { "id": "authors:z24va-45a37", "collection": "authors", "collection_id": "z24va-45a37", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200409-115737860", "type": "book_section", "title": "Comparative analysis of human and mouse development: From zygote to pre-gastrulation", "book_title": "Gastrulation: From Embryonic Pattern to Form", "author": [ { "family_name": "Mol\u00e8", "given_name": "Matteo A.", "clpid": "Mol\u00e8-M-A" }, { "family_name": "Weberling", "given_name": "Antonia", "clpid": "Weberling-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "contributor": [ { "family_name": "Solnica-Krezel", "given_name": "Lilianna", "clpid": "Solnica-Krezel-L" } ], "abstract": "Development of the mammalian embryo begins with formation of the totipotent zygote during fertilization. This initial cell is able to give rise to every embryonic tissue of the developing organism as well as all extra-embryonic lineages, such as the placenta and the yolk sac, which are essential for the initial patterning and support growth of the fetus until birth. As the embryo transits from pre- to post-implantation, major structural and transcriptional changes occur within the embryonic lineage to set up the basis for the subsequent phase of gastrulation. Fine-tuned coordination of cell division, morphogenesis and differentiation is essential to ultimately promote assembly of the future fetus. Here, we review the current knowledge of mammalian development of both mouse and human focusing on morphogenetic processes leading to the onset of gastrulation, when the embryonic anterior-posterior axis becomes established and the three germ layers start to be specified.", "doi": "10.1016/bs.ctdb.2019.10.002", "isbn": "9780128127988", "publisher": "Elsevier", "place_of_publication": "Cambridge, MA", "publication_date": "2020", "pages": "113-138" }, { "id": "authors:z504g-8q339", "collection": "authors", "collection_id": "z504g-8q339", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191216-145145496", "type": "article", "title": "Self-Organization of Mouse Stem Cells into an Extended Potential Blastoid", "author": [ { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-Berna" }, { "family_name": "Cox", "given_name": "Andy L.", "clpid": "Cox-Andy-L" }, { "family_name": "De Jonghe", "given_name": "Joachim", "orcid": "0000-0003-0584-8265", "clpid": "De-Jonghe-Joachim" }, { "family_name": "Bao", "given_name": "Min", "orcid": "0000-0003-0992-9388", "clpid": "Bao-Min" }, { "family_name": "Hollfelder", "given_name": "Florian", "orcid": "0000-0002-1367-6312", "clpid": "Hollfelder-Florian" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Mammalian blastocysts comprise three distinct cell lineages essential for development beyond implantation: the pluripotent epiblast, which generates the future embryo, and surrounding it the extra-embryonic primitive endoderm and the trophectoderm tissues. Embryonic stem cells can reintegrate into embryogenesis but contribute primarily to epiblast lineages. Here, we show that mouse embryonic stem cells cultured under extended pluripotent conditions (EPSCs) can be partnered with trophoblast stem cells to self-organize into blastocyst-like structures with all three embryonic and extra-embryonic lineages. Morphogenetic and transcriptome profiling analyses reveal that these blastocyst-like structures show distinct embryonic-abembryonic axes and primitive endoderm differentiation and can initiate the transition from the pre- to post-implantation egg cylinder morphology in vitro.", "doi": "10.1016/j.devcel.2019.11.014", "pmcid": "PMC10291877", "issn": "1534-5807", "publisher": "Cell Press", "publication": "Developmental Cell", "publication_date": "2019-12-16", "series_number": "6", "volume": "51", "issue": "6", "pages": "698-712" }, { "id": "authors:0es9e-p7c34", "collection": "authors", "collection_id": "0es9e-p7c34", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191111-124015897", "type": "article", "title": "Epigenetic remodelling licences adult cholangiocytes for organoid formation and liver regeneration", "author": [ { "family_name": "Aloia", "given_name": "Luigi", "orcid": "0000-0003-4509-2730", "clpid": "Aloia-L" }, { "family_name": "McKie", "given_name": "Mikel Alexander", "orcid": "0000-0002-1711-4034", "clpid": "McKie-M-A" }, { "family_name": "Vernaz", "given_name": "Gr\u00e9goire", "orcid": "0000-0001-8942-2370", "clpid": "Vernaz-G" }, { "family_name": "Cordero-Espinoza", "given_name": "Luc\u00eda", "clpid": "Cordero-Espinoza-L" }, { "family_name": "Aleksieva", "given_name": "Niya", "clpid": "Aleksieva-N" }, { "family_name": "van den Ameele", "given_name": "Jelle", "orcid": "0000-0002-2744-0810", "clpid": "van-den-Ameele-J" }, { "family_name": "Antonica", "given_name": "Francesco", "orcid": "0000-0001-9359-9689", "clpid": "Antonica-F" }, { "family_name": "Font-Cunill", "given_name": "Berta", "clpid": "Font-Cunill-B" }, { "family_name": "Raven", "given_name": "Alexander", "clpid": "Raven-A" }, { "family_name": "Cigliano", "given_name": "Riccardo Aiese", "clpid": "Cigliano-R-A" }, { "family_name": "Belenguer", "given_name": "German", "clpid": "Belenguer-G" }, { "family_name": "Mort", "given_name": "Richard L.", "orcid": "0000-0002-3566-8217", "clpid": "Mort-R-L" }, { "family_name": "Brand", "given_name": "Andrea H.", "clpid": "Brand-A-H" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Forbes", "given_name": "Stuart J.", "clpid": "Forbes-S-J" }, { "family_name": "Miska", "given_name": "Eric A.", "clpid": "Miska-E-A" }, { "family_name": "Huch", "given_name": "Meritxell", "clpid": "Huch-M" } ], "abstract": "Following severe or chronic liver injury, adult ductal cells (cholangiocytes) contribute to regeneration by restoring both hepatocytes and cholangiocytes. We recently showed that ductal cells clonally expand as self-renewing liver organoids that retain their differentiation capacity into both hepatocytes and ductal cells. However, the molecular mechanisms by which adult ductal-committed cells acquire cellular plasticity, initiate organoids and regenerate the damaged tissue remain largely unknown. Here, we describe that ductal cells undergo a transient, genome-wide, remodelling of their transcriptome and epigenome during organoid initiation and in vivo following tissue damage. TET1-mediated hydroxymethylation licences differentiated ductal cells to initiate organoids and activate the regenerative programme through the transcriptional regulation of stem-cell genes and regenerative pathways including the YAP\u2013Hippo signalling. Our results argue in favour of the remodelling of genomic methylome/hydroxymethylome landscapes as a general mechanism by which differentiated cells exit a committed state in response to tissue damage.", "doi": "10.1038/s41556-019-0402-6", "pmcid": "PMC6940196", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2019-11-04", "series_number": "11", "volume": "21", "issue": "11", "pages": "1321-1333" }, { "id": "authors:zvefc-wps44", "collection": "authors", "collection_id": "zvefc-wps44", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20191017-144439188", "type": "article", "title": "Voices in methods development", "author": [ { "family_name": "Anikeeva", "given_name": "Polina", "clpid": "Anikeeva-P" }, { "family_name": "Ciss\u00e9", "given_name": "Ibrahim I.", "orcid": "0000-0002-8764-1809", "clpid": "Ciss\u00e9-I-I" }, { "family_name": "Jensen", "given_name": "Grant Jay", "orcid": "0000-0003-1556-4864", "clpid": "Jensen-G-J" }, { "family_name": "Van Valen", "given_name": "David", "orcid": "0000-0001-7534-7621", "clpid": "Van-Valen-D" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "To mark the 15th anniversary of Nature Methods, we asked scientists from across diverse fields of basic biology research for their views on the most exciting and essential methodological challenges that their communities are poised to tackle in the near future.", "doi": "10.1038/s41592-019-0585-6", "pmcid": "PMC7075469", "issn": "1548-7091", "publisher": "Nature Publishing Group", "publication": "Nature Methods", "publication_date": "2019-10", "series_number": "10", "volume": "16", "issue": "10", "pages": "945-951" }, { "id": "authors:294ex-kgk14", "collection": "authors", "collection_id": "294ex-kgk14", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200409-115738046", "type": "article", "title": "Morphogenesis of extra-embryonic tissues directs the remodelling of the mouse embryo at implantation", "author": [ { "family_name": "Christodoulou", "given_name": "Neophytos", "orcid": "0000-0003-4338-5847", "clpid": "Christodoulou-N" }, { "family_name": "Weberling", "given_name": "Antonia", "clpid": "Weberling-A" }, { "family_name": "Strathdee", "given_name": "Douglas", "orcid": "0000-0003-2959-4327", "clpid": "Strathdee-D" }, { "family_name": "Anderson", "given_name": "Kurt I.", "orcid": "0000-0002-9324-9598", "clpid": "Anderson-K-I" }, { "family_name": "Timpson", "given_name": "Paul", "clpid": "Timpson-P" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Mammalian embryos change shape dramatically upon implantation. The cellular and molecular mechanism underlying this transition are largely unknown. Here, we show that this transition is directed by cross talk between the embryonic epiblast and the first extra-embryonic tissue, the trophectoderm. Specifically, we show via visualisation of a Cdx2-GFP reporter line and pharmacologically mediated loss and gain of function experiments that the epiblast provides FGF signal that results in differential fate acquisition in the multipotent trophectoderm leading to the formation of a tissue boundary within this tissue. The trophectoderm boundary becomes essential for expansion of the tissue into a multi-layered epithelium. Folding of this multi-layered trophectoderm induces spreading of the second extra-embryonic tissue, the primitive endoderm. Together, these events remodel the pre-implantation embryo into its post-implantation cylindrical shape. Our findings uncover how communication between embryonic and extra-embryonic tissues provides positional cues to drive shape changes in mammalian development during implantation.", "doi": "10.1038/s41467-019-11482-5", "pmcid": "PMC6686005", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2019-08-07", "volume": "10", "pages": "Art. No. 3557" }, { "id": "authors:wwxex-7k660", "collection": "authors", "collection_id": "wwxex-7k660", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111045626", "type": "article", "title": "Concerted cell divisions in embryonic visceral endoderm guide anterior visceral endoderm migration", "author": [ { "family_name": "Antonica", "given_name": "Francesco", "orcid": "0000-0001-9359-9689", "clpid": "Antonica-F" }, { "family_name": "Orietti", "given_name": "Lorenzo Carlo", "clpid": "Orietti-L-C" }, { "family_name": "Mort", "given_name": "Richard Lester", "orcid": "0000-0002-3566-8217", "clpid": "Mort-R-L" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Migration of Anterior Visceral Endoderm (AVE) is a critical symmetry breaking event in the early post-implantation embryo development and is essential for establishing the correct body plan. Despite much effort, cellular and molecular events influencing AVE migration are only partially understood. Here, using time-lapse live imaging of mouse embryos, we demonstrate that cell division in the embryonic visceral endoderm is coordinated with AVE migration. Moreover, we demonstrate that temporal inhibition of FGF signalling during the pre-implantation specification of embryonic visceral endoderm perturbs cell cycle progression, thus affecting AVE migration. These findings demonstrate that coordinated cell cycle progression during the implantation stages of development is important for post-implantation morphogenesis in the mouse embryo.", "doi": "10.1016/j.ydbio.2019.03.016", "pmcid": "PMC6553843", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2019-06-15", "series_number": "2", "volume": "450", "issue": "2", "pages": "132-140" }, { "id": "authors:bjefj-fzn10", "collection": "authors", "collection_id": "bjefj-fzn10", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20200409-115738157", "type": "article", "title": "Self-organization of stem cells into embryos: A window on early mammalian development", "author": [ { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-M-N" }, { "family_name": "Siggia", "given_name": "Eric D.", "orcid": "0000-0001-7482-1854", "clpid": "Siggia-E-D" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Embryonic development is orchestrated by robust and complex regulatory mechanisms acting at different scales of organization. In vivo studies are particularly challenging for mammals after implantation, owing to the small size and inaccessibility of the embryo. The generation of stem cell models of the embryo represents a powerful system with which to dissect this complexity. Control of geometry, modulation of the physical environment, and priming with chemical signals reveal the intrinsic capacity of embryonic stem cells to make patterns. Adding the stem cells for the extraembryonic lineages generates three-dimensional models that are more autonomous from the environment and recapitulate many features of the pre- and postimplantation mouse embryo, including gastrulation. Here, we review the principles of self-organization and how they set cells in motion to create an embryo.", "doi": "10.1126/science.aax0164", "issn": "0036-8075", "publisher": "American Association for the Advancement of Science", "publication": "Science", "publication_date": "2019-06-07", "series_number": "6444", "volume": "364", "issue": "6444", "pages": "948-951" }, { "id": "authors:1vvst-jnr69", "collection": "authors", "collection_id": "1vvst-jnr69", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111045813", "type": "article", "title": "CARM1 and Paraspeckles Regulate Pre-implantation Mouse Embryo Development", "author": [ { "family_name": "Hupalowska", "given_name": "Anna", "clpid": "Hupalowsk-A" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Bedford", "given_name": "Mark T.", "clpid": "Bedford-M-T" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Nuclear architecture has never been carefully examined during early mammalian development at the stages leading to establishment of the embryonic and extra-embryonic lineages. Heterogeneous activity of the methyltransferase CARM1 during these stages results in differential methylation of histone H3R26 to modulate establishment of these two lineages. Here we show that CARM1 accumulates in nuclear granules at the 2- to 4-cell stage transition in the mouse embryo, with the majority corresponding to paraspeckles. The paraspeckle component Neat1 and its partner p54nrb are required for CARM1's association with paraspeckles and for H3R26 methylation. Conversely, CARM1 also influences paraspeckle organization. Depletion of Neat1 or p54nrb results in arrest at the 16- to 32-cell stage, with elevated expression of transcription factor Cdx2, promoting differentiation into the extra-embryonic lineage. This developmental arrest occurs at an earlier stage than following CARM1 depletion, indicating that paraspeckles act upstream of CARM1 but also have additional earlier roles in fate choice.", "doi": "10.1016/j.cell.2018.11.027", "pmcid": "PMC6292842", "issn": "0092-8674", "publisher": "Elsevier", "publication": "Cell", "publication_date": "2018-12-13", "series_number": "7", "volume": "175", "issue": "7", "pages": "1902-1916.e13" }, { "id": "authors:01m59-rt533", "collection": "authors", "collection_id": "01m59-rt533", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111045932", "type": "article", "title": "Debate ethics of embryo models from stem cells", "author": [ { "family_name": "Rivron", "given_name": "Nicolas", "clpid": "Rivron-N" }, { "family_name": "Pera", "given_name": "Martin", "clpid": "Pera-M" }, { "family_name": "Rossant", "given_name": "Janet", "clpid": "Rossant-J" }, { "family_name": "Martinez Arias", "given_name": "Alfonso", "clpid": "Martinez-Arias-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Fu", "given_name": "Jianping", "clpid": "Fu-Jianping" }, { "family_name": "van den Brink", "given_name": "Susanne", "clpid": "van-den-Brink-S" }, { "family_name": "Bredenoord", "given_name": "Annelien", "clpid": "Bredenoord-A" }, { "family_name": "Dondorp", "given_name": "Wybo", "clpid": "Dondorp-W" }, { "family_name": "de Wert", "given_name": "Guido", "clpid": "de-Wert-G" }, { "family_name": "Hyun", "given_name": "Insoo", "clpid": "Hyun-Insoo" }, { "family_name": "Munsie", "given_name": "Megan", "clpid": "Munsie-M" }, { "family_name": "Isasi", "given_name": "Rosario", "clpid": "Isasi-R" } ], "abstract": "[no abstract", "doi": "10.1038/d41586-018-07663-9", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2018-12-13", "series_number": "7735", "volume": "564", "issue": "7735", "pages": "183-185" }, { "id": "authors:9r57j-y4635", "collection": "authors", "collection_id": "9r57j-y4635", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111046061", "type": "article", "title": "Sequential formation and resolution of multiple rosettes drive embryo remodelling after implantation", "author": [ { "family_name": "Christodoulou", "given_name": "Neophytos", "orcid": "0000-0003-4338-5847", "clpid": "Christodoulou-N" }, { "family_name": "Kyprianou", "given_name": "Christos", "orcid": "0000-0001-8516-9195", "clpid": "Kyprianou-C" }, { "family_name": "Weberling", "given_name": "Antonia", "clpid": "Weberling-A" }, { "family_name": "Wang", "given_name": "Ran", "orcid": "0000-0003-4956-5742", "clpid": "Wang-Ran" }, { "family_name": "Cui", "given_name": "Guizhong", "clpid": "Cui-Guizhong" }, { "family_name": "Peng", "given_name": "Guangdun", "orcid": "0000-0002-8586-0637", "clpid": "Peng-Guangdun" }, { "family_name": "Jing", "given_name": "Naihe", "orcid": "0000-0003-1509-6378", "clpid": "Jing-Naihe" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The morphogenetic remodelling of embryo architecture after implantation culminates in pro-amniotic cavity formation. Despite its key importance, how this transformation occurs remains unknown. Here, we apply high-resolution imaging of embryos developing in vivo and in vitro, spatial RNA sequencing and 3D trophoblast stem cell models to determine the sequence and mechanisms of these remodelling events. We show that cavitation of the embryonic tissue is followed by folding of extra-embryonic tissue to mediate the formation of a second extra-embryonic cavity. Concomitantly, at the boundary between embryonic and extra-embryonic tissues, a hybrid 3D rosette forms. Resolution of this rosette enables the embryonic cavity to invade the extra-embryonic tissue. Subsequently, \u03b21-integrin signalling mediates the formation of multiple extra-embryonic 3D rosettes. Podocalyxin exocytosis leads to their polarized resolution, permitting the extension of embryonic and extra-embryonic cavities and their fusion into a unified pro-amniotic cavity. These morphogenetic transformations of embryogenesis reveal a previously unappreciated mechanism for lumen expansion and fusion.", "doi": "10.1038/s41556-018-0211-3", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2018-11", "series_number": "11", "volume": "20", "issue": "11", "pages": "1278-1289" }, { "id": "authors:c26rd-z4628", "collection": "authors", "collection_id": "c26rd-z4628", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-161504340", "type": "book_section", "title": "Delivery of mtZFNs into Early Mouse Embryos", "book_title": "Zinc Finger Proteins: Methods and Protocols", "author": [ { "family_name": "McCann", "given_name": "Beverly J.", "clpid": "McCann-B-J" }, { "family_name": "Cox", "given_name": "Andy", "clpid": "Cox-Andy" }, { "family_name": "Gammage", "given_name": "Payam A.", "clpid": "Gammage-P-A" }, { "family_name": "Stewart", "given_name": "James B.", "clpid": "Stewart-J-B" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Minczuk", "given_name": "Michal", "clpid": "Minczuk-M" } ], "contributor": [ { "family_name": "Liu", "given_name": "Jia", "clpid": "Liu-Jia" } ], "abstract": "Mitochondrial diseases often result from mutations in the mitochondrial genome (mtDNA). In most cases, mutant mtDNA coexists with wild-type mtDNA, resulting in heteroplasmy. One potential future approach to treat heteroplasmic mtDNA diseases is the specific elimination of pathogenic mtDNA mutations, lowering the level of mutant mtDNA below pathogenic thresholds. Mitochondrially targeted zinc-finger nucleases (mtZFNs) have been demonstrated to specifically target and introduce double-strand breaks in mutant mtDNA, facilitating substantial shifts in heteroplasmy. One application of mtZFN technology, in the context of heteroplasmic mtDNA disease, is delivery into the heteroplasmic oocyte or early embryo to eliminate mutant mtDNA, preventing transmission of mitochondrial diseases through the germline. Here we describe a protocol for efficient production of mtZFN mRNA in vitro, and delivery of these into 0.5 dpc mouse embryos to elicit shifts of mtDNA heteroplasmy.", "doi": "10.1007/978-1-4939-8799-3_16", "isbn": "9781493987986", "publisher": "Humana Press", "place_of_publication": "New York, NY", "publication_date": "2018-08-29", "pages": "215-228" }, { "id": "authors:42zvd-2ch71", "collection": "authors", "collection_id": "42zvd-2ch71", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314164", "type": "article", "title": "Deconstructing and reconstructing the mouse and human early embryo", "author": [ { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-M-N" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The emergence of form and function during mammalian embryogenesis is a complex process that involves multiple regulatory levels. The foundations of the body plan are laid throughout the first days of post-implantation development as embryonic stem cells undergo symmetry breaking and initiate lineage specification, in a process that coincides with a global morphological reorganization of the embryo. Here, we review experimental models and how they have shaped our current understanding of the post-implantation mammalian embryo.", "doi": "10.1038/s41556-018-0144-x", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2018-08", "series_number": "8", "volume": "20", "issue": "8", "pages": "878-887" }, { "id": "authors:pwkq4-zzr83", "collection": "authors", "collection_id": "pwkq4-zzr83", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111046182", "type": "article", "title": "Self-assembly of embryonic and two extra-embryonic stem cell types into gastrulating embryo-like structures", "author": [ { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-B" }, { "family_name": "Amadei", "given_name": "Gianluca", "clpid": "Amadei-G" }, { "family_name": "Cox", "given_name": "Andy", "clpid": "Cox-Andy" }, { "family_name": "Wang", "given_name": "Ran", "orcid": "0000-0003-4956-5742", "clpid": "Wang-Ran" }, { "family_name": "Na", "given_name": "Ellen", "orcid": "0000-0003-1395-9707", "clpid": "Na-Ellen" }, { "family_name": "Czukiewska", "given_name": "Sylwia", "clpid": "Czukiewska-S" }, { "family_name": "Chappell", "given_name": "Lia", "clpid": "Chappell-L" }, { "family_name": "Voet", "given_name": "Thierry", "clpid": "Voet-T" }, { "family_name": "Michel", "given_name": "Geert", "clpid": "Michel-G" }, { "family_name": "Jing", "given_name": "Naihe", "orcid": "0000-0003-1509-6378", "clpid": "Jing-Naihe" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Embryonic stem cells can be incorporated into the developing embryo and its germ line, but, when cultured alone, their ability to generate embryonic structures is restricted. They can interact with trophoblast stem cells to generate structures that break symmetry and specify mesoderm, but their development is limited as the epithelial\u2013mesenchymal transition of gastrulation cannot occur. Here, we describe a system that allows assembly of mouse embryonic, trophoblast and extra-embryonic endoderm stem cells into structures that acquire the embryo's architecture with all distinct embryonic and extra-embryonic compartments. Strikingly, such embryo-like structures develop to undertake the epithelial\u2013mesenchymal transition, leading to mesoderm and then definitive endoderm specification. Spatial transcriptomic analyses demonstrate that these morphological transformations are underpinned by gene expression patterns characteristic of gastrulating embryos. This demonstrates the remarkable ability of three stem cell types to self-assemble in vitro into gastrulating embryo-like structures undertaking spatio-temporal events of the gastrulating mammalian embryo.", "doi": "10.1038/s41556-018-0147-7", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2018-08", "series_number": "8", "volume": "20", "issue": "8", "pages": "979-989" }, { "id": "authors:pq4mg-ewf88", "collection": "authors", "collection_id": "pq4mg-ewf88", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314258", "type": "article", "title": "In vitro generation of mouse polarized embryo-like structures from embryonic and trophoblast stem cells", "author": [ { "family_name": "Harrison", "given_name": "Sarah Ellys", "orcid": "0000-0002-1571-2399", "clpid": "Harrison-S-E" }, { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-B" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Mammalian embryogenesis requires the coordination of embryonic and extra-embryonic tissues to enable implantation into the uterus and post-implantation development to establish the body plan. Mouse embryonic stem cells (ESCs) are a useful tool for studying pluripotent embryonic tissue in vitro. However, they cannot undertake correct embryogenesis alone. Many attempts to model the early embryo in vitro involve the aggregation of ESCs into spheroids of variable size and cell number that undertake germ-layer specification but fail to recapitulate the characteristic architecture and arrangement of tissues of the early embryo. Here, we describe a protocol to generate the first embryo-like structures by directing the assembly of mouse ESCs and extra-embryonic trophoblast stem cells (TSCs) in a 3D extracellular matrix (ECM) into structures we call 'polarized embryo-like structures'. By establishing the medium and culture conditions needed to support the growth of both stem cell types simultaneously, embryonic architecture is generated within 4 d of co-culture. This protocol can be performed by those proficient in standard ESC culture techniques and can be used in developmental studies to investigate the interactions between embryonic and extra-embryonic tissues during mammalian development.", "doi": "10.1038/s41596-018-0005-x", "issn": "1754-2189", "publisher": "Nature Publishing Group", "publication": "Nature Protocols", "publication_date": "2018-07", "series_number": "7", "volume": "13", "issue": "7", "pages": "1586-1602" }, { "id": "authors:bszea-1jh11", "collection": "authors", "collection_id": "bszea-1jh11", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111046445", "type": "article", "title": "Tracing the origin of heterogeneity and symmetry breaking in the early mammalian embryo", "author": [ { "family_name": "Chen", "given_name": "Qi", "orcid": "0000-0001-6353-9589", "clpid": "Chen-Qi" }, { "family_name": "Shi", "given_name": "Junchao", "orcid": "0000-0002-1548-5069", "clpid": "Shi-Junchao" }, { "family_name": "Tao", "given_name": "Yi", "orcid": "0000-0002-0749-6614", "clpid": "Tao-Yi" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "A fundamental question in developmental and stem cell biology concerns the origin and nature of signals that initiate asymmetry leading to pattern formation and self-organization. Instead of having prominent pre-patterning determinants as present in model organisms (worms, sea urchin, frog), we propose that the mammalian embryo takes advantage of more subtle cues such as compartmentalized intracellular reactions that generate micro-scale inhomogeneity, which is gradually amplified over several cellular generations to drive pattern formation while keeping developmental plasticity. It is therefore possible that by making use of compartmentalized information followed by its amplification, mammalian embryos would follow general principle of development found in other organisms in which the spatial cue is more robustly presented.", "doi": "10.1038/s41467-018-04155-2", "pmcid": "PMC5940674", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2018-05-08", "volume": "9", "pages": "Art. No. 1819" }, { "id": "authors:dqxbd-gy925", "collection": "authors", "collection_id": "dqxbd-gy925", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111046546", "type": "article", "title": "RASSF1A uncouples Wnt from Hippo signalling and promotes YAP mediated differentiation via p73", "author": [ { "family_name": "Papaspyropoulos", "given_name": "Angelos", "clpid": "Papaspyropoulos-A" }, { "family_name": "Bradley", "given_name": "Leanne", "clpid": "Bradley-L" }, { "family_name": "Thapa", "given_name": "Asmita", "clpid": "Thapa-A" }, { "family_name": "Leung", "given_name": "Chuen Yan", "clpid": "Leung-Chuen-Yan" }, { "family_name": "Toskas", "given_name": "Konstantinos", "clpid": "Toskas-K" }, { "family_name": "Koennig", "given_name": "Delia", "clpid": "Koennig-D" }, { "family_name": "Pefani", "given_name": "Dafni-Eleftheria", "clpid": "Pefani-D-E" }, { "family_name": "Raso", "given_name": "Cinzia", "clpid": "Raso-C" }, { "family_name": "Grou", "given_name": "Claudia", "clpid": "Grou-C" }, { "family_name": "Hamilton", "given_name": "Garth", "clpid": "Hamilton-G" }, { "family_name": "Vlahov", "given_name": "Nikola", "clpid": "Vlahov-N" }, { "family_name": "Grawenda", "given_name": "Anna", "clpid": "Grawenda-A" }, { "family_name": "Haider", "given_name": "Syed", "clpid": "Haider-S" }, { "family_name": "Chauhan", "given_name": "Jagat", "orcid": "0000-0003-0330-1901", "clpid": "Chauhan-J" }, { "family_name": "Buti", "given_name": "Ludovico", "clpid": "Buti-L" }, { "family_name": "Kanapin", "given_name": "Alexander", "orcid": "0000-0001-9802-5297", "clpid": "Kanapin-A" }, { "family_name": "Lu", "given_name": "Xin", "clpid": "Lu-Xin" }, { "family_name": "Buffa", "given_name": "Francesca", "orcid": "0000-0003-0409-406X", "clpid": "Buffa-F" }, { "family_name": "Dianov", "given_name": "Grigory", "clpid": "Dianov-G" }, { "family_name": "von Kriegsheim", "given_name": "Alex", "clpid": "von-Kriegsheim-A" }, { "family_name": "Matallanas", "given_name": "David", "clpid": "Matallanas-D" }, { "family_name": "Samsonova", "given_name": "Anastasia", "clpid": "Samsonova-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "O'Neill", "given_name": "Eric", "orcid": "0000-0002-0060-6278", "clpid": "O'Neill-E" } ], "abstract": "Transition from pluripotency to differentiation is a pivotal yet poorly understood developmental step. Here, we show that the tumour suppressor RASSF1A is a key player driving the early specification of cell fate. RASSF1A acts as a natural barrier to stem cell self-renewal and iPS cell generation, by switching YAP from an integral component in the \u03b2-catenin-TCF pluripotency network to a key factor that promotes differentiation. We demonstrate that epigenetic regulation of the Rassf1A promoter maintains stemness by allowing a quaternary association of YAP\u2013TEAD and \u03b2-catenin\u2013TCF3 complexes on the Oct4 distal enhancer. However, during differentiation, promoter demethylation allows GATA1-mediated RASSF1A expression which prevents YAP from contributing to the TEAD/\u03b2-catenin\u2013TCF3 complex. Simultaneously, we find that RASSF1A promotes a YAP\u2013p73 transcriptional programme that enables differentiation. Together, our findings demonstrate that RASSF1A mediates transcription factor selection of YAP in stem cells, thereby acting as a functional \"switch\" between pluripotency and initiation of differentiation.", "doi": "10.1038/s41467-017-02786-5", "pmcid": "PMC5789973", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2018-01-30", "volume": "9", "pages": "Art. No. 424" }, { "id": "authors:dfy59-xcs95", "collection": "authors", "collection_id": "dfy59-xcs95", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111046720", "type": "article", "title": "Cyclin B1 is essential for mitosis in mouse embryos, and its nuclear export sets the time for mitosis", "author": [ { "family_name": "Strauss", "given_name": "Bernhard", "clpid": "Strauss-Bernhard" }, { "family_name": "Harrison", "given_name": "Andrew", "orcid": "0000-0002-3778-8415", "clpid": "Harrison-Andrew" }, { "family_name": "Almeida Coelho", "given_name": "Paula", "orcid": "0000-0003-0614-7575", "clpid": "Almeida-Coelho-Paula" }, { "family_name": "Yata", "given_name": "Keiko", "orcid": "0000-0001-7648-4898", "clpid": "Yata-Keiko" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Pines", "given_name": "Jonathon", "orcid": "0000-0002-5227-6004", "clpid": "Pines-Jonathon" } ], "abstract": "There is remarkable redundancy between the Cyclin\u2013Cdk complexes that comprise the cell cycle machinery. None of the mammalian A-, D-, or E-type cyclins are required in development until implantation, and only Cdk1 is essential for early cell divisions. Cyclin B1 is essential for development, but whether it is required for cell division is contentious. Here, we used a novel imaging approach to analyze Cyclin B1\u2013null embryos from fertilization onward. We show that Cyclin B1\u2212/\u2212 embryos arrest in G2 phase after just two divisions. This is the earliest arrest of any Cyclin known and places Cyclin B1 with cdk1 as the essential regulators of the cell cycle. We reintroduced mutant proteins into this genetically null background to determine why Cyclin B1 is constantly exported from the nucleus. We found that Cyclin B1 must be exported from the nucleus for the cell to prevent premature entry to mitosis, and retaining Cyclin B1\u2013Cdk1 at the plasma membrane precludes entry to mitosis.", "doi": "10.1083/jcb.201612147", "pmcid": "PMC5748970", "issn": "0021-9525", "publisher": "Rockefeller University Press", "publication": "Journal of Cell Biology", "publication_date": "2018-01-02", "series_number": "1", "volume": "217", "issue": "1", "pages": "179-193" }, { "id": "authors:apt5s-7zv34", "collection": "authors", "collection_id": "apt5s-7zv34", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-111046314", "type": "article", "title": "Pluripotent state transitions coordinate morphogenesis in mouse and human embryos", "author": [ { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-Marta-N" }, { "family_name": "Scialdone", "given_name": "Antonio", "clpid": "Scialdone-Antonio" }, { "family_name": "Skorupska", "given_name": "Natalia", "clpid": "Skorupska-Natalia" }, { "family_name": "Weberling", "given_name": "Antonia", "clpid": "Weberling-Antonia" }, { "family_name": "Recher", "given_name": "Gaelle", "orcid": "0000-0001-9004-0118", "clpid": "Recher-Gaelle" }, { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-Agnieszka" }, { "family_name": "Devito", "given_name": "Liani G.", "clpid": "Devito-Liani-G" }, { "family_name": "Noli", "given_name": "Laila", "clpid": "Noli-Laila" }, { "family_name": "Macaulay", "given_name": "Iain C.", "clpid": "Macaulay-Iain-C" }, { "family_name": "Buecker", "given_name": "Christa", "clpid": "Buecker-Christa" }, { "family_name": "Khalaf", "given_name": "Yakoub", "clpid": "Khalaf-Yakoub" }, { "family_name": "Ilic", "given_name": "Dusko", "clpid": "Ilic-Dusko" }, { "family_name": "Voet", "given_name": "Thierry", "clpid": "Voet-Thierry" }, { "family_name": "Marioni", "given_name": "John C.", "clpid": "Marioni-John-C" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The foundations of mammalian development lie in a cluster of embryonic epiblast stem cells. In response to extracellular matrix signalling, these cells undergo epithelialization and create an apical surface in contact with a cavity, a fundamental event for all subsequent development. Concomitantly, epiblast cells transit through distinct pluripotent states, before lineage commitment at gastrulation. These pluripotent states have been characterized at the molecular level5, but their biological importance remains unclear. Here we show that exit from an unrestricted naive pluripotent state is required for epiblast epithelialization and generation of the pro-amniotic cavity in mouse embryos. Embryonic stem cells locked in the naive state are able to initiate polarization but fail to undergo lumenogenesis. Mechanistically, exit from naive pluripotency activates an Oct4-governed transcriptional program that results in expression of glycosylated sialomucin proteins and the vesicle tethering and fusion events of lumenogenesis. Similarly, exit of epiblasts from naive pluripotency in cultured human post-implantation embryos triggers amniotic cavity formation and developmental progression. Our results add tissue-level architecture as a new criterion for the characterization of different pluripotent states, and show the relevance of transitions between these states during development of the mammalian embryo.", "doi": "10.1038/nature24675", "pmcid": "PMC5768241", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2017-12-14", "series_number": "7684", "volume": "552", "issue": "7684", "pages": "239-243" }, { "id": "authors:63361-5pn82", "collection": "authors", "collection_id": "63361-5pn82", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-162605585", "type": "article", "title": "Plk4 and Aurora A cooperate in the initiation of acentriolar spindle assembly in mammalian oocytes", "author": [ { "family_name": "Bury", "given_name": "Leah", "orcid": "0000-0002-6218-2837", "clpid": "Bury-Leah" }, { "family_name": "Coelho", "given_name": "Paula A.", "orcid": "0000-0003-0614-7575", "clpid": "Coelho-Paula-A" }, { "family_name": "Simeone", "given_name": "Angela", "clpid": "Simeone-Angela" }, { "family_name": "Ferries", "given_name": "Samantha", "clpid": "Ferries-Samantha" }, { "family_name": "Eyers", "given_name": "Claire E.", "orcid": "0000-0002-3223-5926", "clpid": "Eyers-Claire-E" }, { "family_name": "Eyers", "given_name": "Patrick A.", "orcid": "0000-0002-9220-2966", "clpid": "Eyers-Patrick-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" } ], "abstract": "Establishing the bipolar spindle in mammalian oocytes after their prolonged arrest is crucial for meiotic fidelity and subsequent development. In contrast to somatic cells, the first meiotic spindle assembles in the absence of centriole-containing centrosomes. Ran-GTP can promote microtubule nucleation near chromatin, but additional unidentified factors are postulated for the activity of multiple acentriolar microtubule organizing centers in the oocyte. We now demonstrate that partially overlapping, nonredundant functions of Aurora A and Plk4 kinases contribute to initiate acentriolar meiosis I spindle formation. Loss of microtubule nucleation after simultaneous chemical inhibition of both kinases can be significantly rescued by drug-resistant Aurora A alone. Drug-resistant Plk4 can enhance Aurora A\u2013mediated rescue, and, accordingly, Plk4 can phosphorylate and potentiate the activity of Aurora A in vitro. Both kinases function distinctly from Ran, which amplifies microtubule growth. We conclude that Aurora A and Plk4 are rate-limiting factors contributing to microtubule growth as the acentriolar oocyte resumes meiosis.", "doi": "10.1083/jcb.201606077", "pmcid": "PMC5674873", "issn": "0021-9525", "publisher": "Rockefeller University Press", "publication": "Journal of Cell Biology", "publication_date": "2017-11-06", "series_number": "11", "volume": "216", "issue": "11", "pages": "3571-3590" }, { "id": "authors:jk61y-jwv02", "collection": "authors", "collection_id": "jk61y-jwv02", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190408-161923858", "type": "article", "title": "Revisiting the Warnock rule", "author": [ { "family_name": "Hurlbut", "given_name": "J. Benjamin", "clpid": "Hurlbut-J-B" }, { "family_name": "Hyun", "given_name": "Insoo", "clpid": "Hyun-Insoo" }, { "family_name": "Levine", "given_name": "Aaron D.", "clpid": "Levine-A-D" }, { "family_name": "Lovell-Badge", "given_name": "Robin", "clpid": "Lovell-Badge-Robin" }, { "family_name": "Lunshof", "given_name": "Jeantine E.", "clpid": "Lunshof-J-E" }, { "family_name": "Matthews", "given_name": "Kirstin R. W.", "clpid": "Matthews-K-R-W" }, { "family_name": "Mills", "given_name": "Peter", "clpid": "Mills-P" }, { "family_name": "Murdoch", "given_name": "Alison", "clpid": "Murdoch-A" }, { "family_name": "Pera", "given_name": "Martin F.", "clpid": "Pera-M-F" }, { "family_name": "Scott", "given_name": "Christopher Thomas", "clpid": "Scott-C-T" }, { "family_name": "Tizzard", "given_name": "Juliet", "clpid": "Tizzard-J" }, { "family_name": "Warnock", "given_name": "Mary", "clpid": "Warnock-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Zhou", "given_name": "Qi", "clpid": "Zhou-Qi" }, { "family_name": "Zoloth", "given_name": "Laurie", "clpid": "Zoloth-L" } ], "abstract": "Is it time to reassess the 14-day rule for human embryo research? \n\nThe seminal 1984 Warnock Report established that research on human embryos should be limited to the first 14 days of development (Box 1). Since that time, the rule has been broadly adopted and adhered to across the research community. With the introduction of new methodologies into human embryology, however, our ability to culture human embryos in vitro has progressed rapidly, to the point where we now are reaching the 14-day Rubicon. In August 2016, two groups in the UK and in the US reported experiments on human embryos that were sustained in culture for 12\u201313 days after fertilization. To comply with British law, the UK lab destroyed its embryo on the 13th day. In the following article, Nature Biotechnology brings together a group of experts to discuss whether, in the light of these advances, it is now time to reassess the 14-day rule.", "doi": "10.1038/nbt.4015", "issn": "1087-0156", "publisher": "Nature Publishing Group", "publication": "Nature Biotechnology", "publication_date": "2017-11", "series_number": "11", "volume": "35", "issue": "11", "pages": "1029-1042" }, { "id": "authors:nqbbt-pzb03", "collection": "authors", "collection_id": "nqbbt-pzb03", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314469", "type": "article", "title": "Actomyosin polarisation through PLC-PKC triggers symmetry breaking of the mouse embryo", "author": [ { "family_name": "Zhu", "given_name": "Meng", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Leung", "given_name": "Chuen Yan", "clpid": "Leung-Chuen-Yan" }, { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-M-N" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Establishment of cell polarity in the mammalian embryo is fundamental for the first cell fate decision that sets aside progenitor cells for both the new organism and the placenta. Yet the sequence of events and molecular mechanism that trigger this process remain unknown. Here, we show that de novo polarisation of the mouse embryo occurs in two distinct phases at the 8-cell stage. In the first phase, an apical actomyosin network is formed. This is a pre-requisite for the second phase, in which the Par complex localises to the apical domain, excluding actomyosin and forming a mature apical cap. Using a variety of approaches, we also show that phospholipase C-mediated PIP_2 hydrolysis is necessary and sufficient to trigger the polarisation of actomyosin through the Rho-mediated recruitment of myosin II to the apical cortex. Together, these results reveal the molecular framework that triggers de novo polarisation of the mouse embryo.", "doi": "10.1038/s41467-017-00977-8", "pmcid": "PMC5640629", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2017-10-13", "volume": "8", "pages": "Art. No. 921" }, { "id": "authors:0ghms-bzc50", "collection": "authors", "collection_id": "0ghms-bzc50", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314558", "type": "article", "title": "Delayed APC/C activation extends the first mitosis of mouse embryos", "author": [ { "family_name": "Ajduk", "given_name": "Anna", "clpid": "Ajduk-A" }, { "family_name": "Strauss", "given_name": "Bernhard", "clpid": "Strauss-B" }, { "family_name": "Pines", "given_name": "Jonathon", "orcid": "0000-0002-5227-6004", "clpid": "Pines-J" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The correct temporal regulation of mitosis underpins genomic stability because it ensures the alignment of chromosomes on the mitotic spindle that is required for their proper segregation to the two daughter cells. Crucially, sister chromatid separation must be delayed until all the chromosomes have attached to the spindle; this is achieved by the Spindle Assembly Checkpoint (SAC) that inhibits the Anaphase Promoting Complex/Cyclosome (APC/C) ubiquitin ligase. In many species the first embryonic M-phase is significantly prolonged compared to the subsequent divisions, but the reason behind this has remained unclear. Here, we show that the first M-phase in the mouse embryo is significantly extended due to a delay in APC/C activation. Unlike in somatic cells, where the APC/C first targets cyclin A2 for degradation at nuclear envelope breakdown (NEBD), we find that in zygotes cyclin A2 remains stable for a significant period of time after NEBD. Our findings that the SAC prevents cyclin A2 degradation, whereas over-expressed Plk1 stimulates it, support our conclusion that the delay in cyclin A2 degradation is caused by low APC/C activity. As a consequence of delayed APC/C activation cyclin B1 stability in the first mitosis is also prolonged, leading to the unusual length of the first M-phase.", "doi": "10.1038/s41598-017-09526-1", "pmcid": "PMC5575289", "issn": "2045-2322", "publisher": "Nature Publishing Group", "publication": "Scientific Reports", "publication_date": "2017-08-29", "volume": "7", "pages": "Art. No. 9682" }, { "id": "authors:p040v-0yt27", "collection": "authors", "collection_id": "p040v-0yt27", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314644", "type": "article", "title": "The chromatin modifier Satb1 regulates cell fate through Fgf signalling in the early mouse embryo", "author": [ { "family_name": "Goolam", "given_name": "Mubeen", "clpid": "Goolam-G" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The separation of embryonic from extra-embryonic tissues within the inner cell mass to generate the epiblast (EPI), which will form the new organism, from the primitive endoderm (PE), which will form the yolk sac, is a crucial developmental decision. Here, we identify a chromatin modifier, Satb1, with a distinct role in this decision. Satb1 is differentially expressed within 16-cell-stage embryos, with higher expression levels in the inner cell mass progenitor cells. Depleting Satb1 increases the number of EPI cells at the expense of PE. This phenotype can be rescued by simultaneous depletion of both Satb1 and Satb2, owing to their antagonistic effect on the pluripotency regulator Nanog. Consequently, increasing Satb1 expression leads to differentiation into PE and a decrease in EPI, as a result of the modulation of expression of several pluripotency- and differentiation-related genes by Satb1. Finally, we show that Satb1 is a downstream target of the Fgf signalling pathway, linking chromatin modification and Fgf signalling. Together, these results identify a role for Satb1 in the lineage choice between pluripotency and differentiation and further our understanding of early embryonic lineage segregation.", "doi": "10.1242/dev.144139", "pmcid": "PMC5399666", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "2017-04-15", "series_number": "8", "volume": "144", "issue": "8", "pages": "1450-1461" }, { "id": "authors:sbxa6-v1f02", "collection": "authors", "collection_id": "sbxa6-v1f02", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314754", "type": "article", "title": "Assembly of embryonic and extraembryonic stem cells to mimic embryogenesis in vitro", "author": [ { "family_name": "Harrison", "given_name": "Sarah Ellys", "orcid": "0000-0002-1571-2399", "clpid": "Harrison-S-E" }, { "family_name": "Sozen", "given_name": "Berna", "orcid": "0000-0001-5834-5819", "clpid": "Sozen-B" }, { "family_name": "Christodoulou", "given_name": "Neophytos", "orcid": "0000-0003-4338-5847", "clpid": "Christodoulou-N" }, { "family_name": "Kyprianou", "given_name": "Christos", "orcid": "0000-0001-8516-9195", "clpid": "Kyprianou-C" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Mammalian embryogenesis requires intricate interactions between embryonic and extraembryonic tissues to orchestrate and coordinate morphogenesis with changes in developmental potential. Here, we combined mouse embryonic stem cells (ESCs) and extraembryonic trophoblast stem cells (TSCs) in a three-dimensional scaffold to generate structures whose morphogenesis is markedly similar to that of natural embryos. By using genetically modified stem cells and specific inhibitors, we show that embryogenesis of ESC- and TSC-derived embryos\u2014ETS-embryos\u2014depends on cross-talk involving Nodal signaling. When ETS-embryos develop, they spontaneously initiate expression of mesoderm and primordial germ cell markers asymmetrically on the embryonic and extraembryonic border, in response to Wnt and BMP signaling. Our study demonstrates the ability of distinct stem cell types to self-assemble in vitro to generate embryos whose morphogenesis, architecture, and constituent cell types resemble those of natural embryos.", "doi": "10.1126/science.aal1810", "issn": "0036-8075", "publisher": "American Association for the Advancement of Science", "publication": "Science", "publication_date": "2017-04-14", "series_number": "6334", "volume": "356", "issue": "6334", "pages": "Art. No. eaal1810" }, { "id": "authors:v9ad4-qyj76", "collection": "authors", "collection_id": "v9ad4-qyj76", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190409-122829890", "type": "article", "title": "Andrzej K. Tarkowski 1933\u20132016", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "With Andrzej Tarkowski's death we lose the father of modern mammalian embryology, whose desire to understand early development brought out a genius for using simple tools and elegant experiments to give true insights into the earliest events of our lives.", "doi": "10.1038/ncb3446", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2016-12", "series_number": "12", "volume": "18", "issue": "12", "pages": "1261" }, { "id": "authors:eth65-08f56", "collection": "authors", "collection_id": "eth65-08f56", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190409-123144282", "type": "article", "title": "Polarity and cell division orientation in the cleavage embryo: from worm to human", "author": [ { "family_name": "Ajduk", "given_name": "Anna", "clpid": "Ajduk-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Cleavage is a period after fertilization, when a 1-cell embryo starts developing into a multicellular organism. Due to a series of mitotic divisions, the large volume of a fertilized egg is divided into numerous smaller, nucleated cells\u2014blastomeres. Embryos of different phyla divide according to different patterns, but molecular mechanism of these early divisions remains surprisingly conserved. In the present paper, we describe how polarity cues, cytoskeleton and cell-to-cell communication interact with each other to regulate orientation of the early embryonic division planes in model animals such as Caenorhabditis elegans, Drosophila and mouse. We focus particularly on the Par pathway and the actin-driven cytoplasmic flows that accompany it. We also describe a unique interplay between Par proteins and the Hippo pathway in cleavage mammalian embryos. Moreover, we discuss the potential meaning of polarity, cytoplasmic dynamics and cell-to-cell communication as quality biomarkers of human embryos.", "doi": "10.1093/molehr/gav068", "pmcid": "PMC5062000", "issn": "1360-9947", "publisher": "Oxford University Press", "publication": "Molecular Human Reproduction", "publication_date": "2016-10", "series_number": "10", "volume": "22", "issue": "10", "pages": "691-703" }, { "id": "authors:yzw1m-ktr05", "collection": "authors", "collection_id": "yzw1m-ktr05", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190409-140137785", "type": "book_section", "title": "Polarity in Cell-Fate Acquisition in the Early Mouse Embryo", "book_title": "Mammalian Preimplantation Development", "author": [ { "family_name": "Leung", "given_name": "C. Y.", "clpid": "Leung-Chuen-Yan" }, { "family_name": "Zhu", "given_name": "M.", "orcid": "0000-0001-6157-8840", "clpid": "Zhu-Meng" }, { "family_name": "Zernicka-Goetz", "given_name": "M.", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "contributor": [ { "family_name": "DePamphilis", "given_name": "Melvin L.", "clpid": "DePamphilis-M-L" } ], "abstract": "Establishing polarity is a fundamental part of embryogenesis and can be traced back to the earliest developmental stages. It can be achieved in one of two ways: through the preexisting polarization of germ cells before fertilization or via symmetry breaking after fertilization. In mammals, it seems to be the latter, and we will discuss the various cytological and molecular events that lead up to this event, its mechanisms and the consequences. In mammals, the first polarization event occurs in the preimplantation period, when the embryo is but a cluster of cells, free-floating in the oviduct. This provides a unique, autonomous system to study the de novo polarization that is essential to life. In this review, we will cover modern and past studies on the polarization of the early embryo, using the mouse as a model system, as well as hypothesizing the potential implications and functions of the biological events involved.", "doi": "10.1016/bs.ctdb.2016.04.008", "isbn": "9780128014288", "publisher": "Elsevier", "place_of_publication": "New York, NY", "publication_date": "2016-06-07", "pages": "203-234" }, { "id": "authors:v1y8n-fmp69", "collection": "authors", "collection_id": "v1y8n-fmp69", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314855", "type": "article", "title": "Self-organization of the human embryo in the absence of maternal tissues", "author": [ { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-M-N" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Vuoristo", "given_name": "Sanna", "clpid": "Vuoristo-S" }, { "family_name": "Recher", "given_name": "Gaelle", "orcid": "0000-0001-9004-0118", "clpid": "Recher-G" }, { "family_name": "Hupalowska", "given_name": "Anna", "clpid": "Hupalowsk-A" }, { "family_name": "Bolton", "given_name": "Virginia", "clpid": "Bolton-V" }, { "family_name": "Fogarty", "given_name": "Norah M. E.", "clpid": "Fogarty-N-M-E" }, { "family_name": "Campbell", "given_name": "Alison", "clpid": "Campbell-A" }, { "family_name": "Devito", "given_name": "Liani G.", "clpid": "Devito-L-G" }, { "family_name": "Ilic", "given_name": "Dusko", "clpid": "Ilic-D" }, { "family_name": "Khalaf", "given_name": "Yakoub", "clpid": "Khalaf-Y" }, { "family_name": "Niakan", "given_name": "Kathy K.", "clpid": "Niakan-K-K" }, { "family_name": "Fishel", "given_name": "Simon", "clpid": "Fishel-S" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Remodelling of the human embryo at implantation is indispensable for successful pregnancy. Yet it has remained mysterious because of the experimental hurdles that beset the study of this developmental phase. Here, we establish an in vitro system to culture human embryos through implantation stages in the absence of maternal tissues and reveal the key events of early human morphogenesis. These include segregation of the pluripotent embryonic and extra-embryonic lineages, and morphogenetic rearrangements leading to generation of a bilaminar disc, formation of a pro-amniotic cavity within the embryonic lineage, appearance of the prospective yolk sac, and trophoblast differentiation. Using human embryos and human pluripotent stem cells, we show that the reorganization of the embryonic lineage is mediated by cellular polarization leading to cavity formation. Together, our results indicate that the critical remodelling events at this stage of human development are embryo-autonomous, highlighting the remarkable and unanticipated self-organizing properties of human embryos.", "doi": "10.1038/ncb3347", "pmcid": "PMC5049689", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2016-06", "series_number": "6", "volume": "18", "issue": "6", "pages": "700-708" }, { "id": "authors:p86sa-3sj31", "collection": "authors", "collection_id": "p86sa-3sj31", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170314964", "type": "article", "title": "Self-organization of the in vitro attached human embryo", "author": [ { "family_name": "Deglincerti", "given_name": "Alessia", "clpid": "Deglincerti-A" }, { "family_name": "Croft", "given_name": "Gist F.", "clpid": "Croft-G-F" }, { "family_name": "Pietila", "given_name": "Lauren N.", "clpid": "Pietila-L-N" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Siggia", "given_name": "Eric D.", "clpid": "Siggia-E-D" }, { "family_name": "Brivanlou", "given_name": "Ali H.", "clpid": "Brivanlou-B-H" } ], "abstract": "Implantation of the blastocyst is a developmental milestone in mammalian embryonic development. At this time, a coordinated program of lineage diversification, cell-fate specification, and morphogenetic movements establishes the generation of extra-embryonic tissues and the embryo proper, and determines the conditions for successful pregnancy and gastrulation. Despite its basic and clinical importance, this process remains mysterious in humans. Here we report the use of a novel in vitro system1,2 to study the post-implantation development of the human embryo. We unveil the self-organizing abilities and autonomy of in vitro attached human embryos. We find human-specific molecular signatures of early cell lineage, timing, and architecture. Embryos display key landmarks of normal development, including epiblast expansion, lineage segregation, bi-laminar disc formation, amniotic and yolk sac cavitation, and trophoblast diversification. Our findings highlight the species-specificity of these developmental events and provide a new understanding of early human embryonic development beyond the blastocyst stage. In addition, our study establishes a new model system relevant to early human pregnancy loss. Finally, our work will also assist in the rational design of differentiation protocols of human embryonic stem cells to specific cell types for disease modelling and cell replacement therapy.", "doi": "10.1038/nature17948", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2016-05", "series_number": "7602", "volume": "533", "issue": "7602", "pages": "251-254" }, { "id": "authors:2m6bp-t1431", "collection": "authors", "collection_id": "2m6bp-t1431", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315353", "type": "article", "title": "The BAF chromatin remodelling complex is an epigenetic regulator of lineage specification in the early mouse embryo", "author": [ { "family_name": "Panamarova", "given_name": "Maryna", "clpid": "Panamarova-M" }, { "family_name": "Cox", "given_name": "Andy", "clpid": "Cox-Andy" }, { "family_name": "Wicher", "given_name": "Krzysztof B.", "clpid": "Wicher-K-B" }, { "family_name": "Butler", "given_name": "Richard", "clpid": "Butler-R" }, { "family_name": "Bulgakova", "given_name": "Natalia", "clpid": "Bulgakova-N" }, { "family_name": "Jeon", "given_name": "Shin", "clpid": "Jeon-Shin" }, { "family_name": "Rosen", "given_name": "Barry", "clpid": "Rosen-B" }, { "family_name": "Seong", "given_name": "Rho H.", "clpid": "Seong-Rho-H" }, { "family_name": "Skarnes", "given_name": "William", "clpid": "Skarnes-W" }, { "family_name": "Crabtree", "given_name": "Gerald", "clpid": "Crabtree-G-R" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Dynamic control of gene expression is essential for the development of a totipotent zygote into an embryo with defined cell lineages. The accessibility of genes responsible for cell specification to transcriptional machinery is dependent on chromatin remodelling complexes such as the SWI\\SNF (BAF) complex. However, the role of the BAF complex in early mouse development has remained unclear. Here, we demonstrate that BAF155, a major BAF complex subunit, regulates the assembly of the BAF complex in vivo and regulates lineage specification of the mouse blastocyst. We find that associations of BAF155 with other BAF complex subunits become enriched in extra-embryonic lineages just prior to implantation. This enrichment is attributed to decreased mobility of BAF155 in extra-embryonic compared with embryonic lineages. Downregulation of BAF155 leads to increased expression of the pluripotency marker Nanog and its ectopic expression in extra-embryonic lineages, whereas upregulation of BAF155 leads to the upregulation of differentiation markers. Finally, we show that the arginine methyltransferase CARM1 methylates BAF155, which differentially influences assembly of the BAF complex between the lineages and the expression of pluripotency markers. Together, our results indicate a novel role of BAF-dependent chromatin remodelling in mouse development via regulation of lineage specification.", "doi": "10.1242/dev.131961", "pmcid": "PMC4852518", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "2016-04-15", "series_number": "8", "volume": "143", "issue": "8", "pages": "1271-1283" }, { "id": "authors:g873b-0xn91", "collection": "authors", "collection_id": "g873b-0xn91", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315061", "type": "article", "title": "Mouse model of chromosome mosaicism reveals lineage-specific depletion of aneuploid cells and normal developmental potential", "author": [ { "family_name": "Bolton", "given_name": "Helen", "clpid": "Bolton-H" }, { "family_name": "Graham", "given_name": "Sarah J. L.", "clpid": "Graham-S-J-L" }, { "family_name": "Van der Aa", "given_name": "Niels", "clpid": "Van-der-Aa-N" }, { "family_name": "Kumar", "given_name": "Parveen", "clpid": "Kumar-P" }, { "family_name": "Theunis", "given_name": "Koen", "clpid": "Theunis-K" }, { "family_name": "Fernandez Gallardo", "given_name": "Elia", "clpid": "Fernandez-Gallardo-E" }, { "family_name": "Voet", "given_name": "Thierry", "clpid": "Voet-T" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Most human pre-implantation embryos are mosaics of euploid and aneuploid cells. To determine the fate of aneuploid cells and the developmental potential of mosaic embryos, here we generate a mouse model of chromosome mosaicism. By treating embryos with a spindle assembly checkpoint inhibitor during the four- to eight-cell division, we efficiently generate aneuploid cells, resulting in embryo death during peri-implantation development. Live-embryo imaging and single-cell tracking in chimeric embryos, containing aneuploid and euploid cells, reveal that the fate of aneuploid cells depends on lineage: aneuploid cells in the fetal lineage are eliminated by apoptosis, whereas those in the placental lineage show severe proliferative defects. Overall, the proportion of aneuploid cells is progressively depleted from the blastocyst stage onwards. Finally, we show that mosaic embryos have full developmental potential, provided they contain sufficient euploid cells, a finding of significance for the assessment of embryo vitality in the clinic.", "doi": "10.1038/ncomms11165", "pmcid": "PMC4820631", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2016-03-29", "volume": "7", "pages": "Art. No. 11165" }, { "id": "authors:vqt7x-wtv82", "collection": "authors", "collection_id": "vqt7x-wtv82", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315162", "type": "article", "title": "Heterogeneity in Oct4 and Sox2 Targets Biases Cell Fate in 4-Cell Mouse Embryos", "author": [ { "family_name": "Goolam", "given_name": "Mubeen", "clpid": "Goolam-G" }, { "family_name": "Scialdone", "given_name": "Antonio", "clpid": "Scialdone-A" }, { "family_name": "Graham", "given_name": "Sarah J. L.", "clpid": "Graham-S-J-L" }, { "family_name": "Macaulay", "given_name": "Iain C.", "clpid": "Macaulay-I-C" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Hupalowska", "given_name": "Anna", "clpid": "Hupalowsk-A" }, { "family_name": "Voet", "given_name": "Thierry", "clpid": "Voet-T" }, { "family_name": "Marioni", "given_name": "John C.", "clpid": "Marioni-J-C" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The major and essential objective of pre-implantation development is to establish embryonic and extra-embryonic cell fates. To address when and how this fundamental process is initiated in mammals, we characterize transcriptomes of all individual cells throughout mouse pre-implantation development. This identifies targets of master pluripotency regulators Oct4 and Sox2 as being highly heterogeneously expressed between blastomeres of the 4-cell embryo, with Sox21 showing one of the most heterogeneous expression profiles. Live-cell tracking demonstrates that cells with decreased Sox21 yield more extra-embryonic than pluripotent progeny. Consistently, decreasing Sox21 results in premature upregulation of the differentiation regulator Cdx2, suggesting that Sox21 helps safeguard pluripotency. Furthermore, Sox21 is elevated following increased expression of the histone H3R26-methylase CARM1 and is lowered following CARM1 inhibition, indicating the importance of epigenetic regulation. Therefore, our results indicate that heterogeneous gene expression, as early as the 4-cell stage, initiates cell-fate decisions by modulating the balance of pluripotency and differentiation.", "doi": "10.1016/j.cell.2016.01.047", "pmcid": "PMC4819611", "issn": "0092-8674", "publisher": "Elsevier", "publication": "Cell", "publication_date": "2016-03-24", "series_number": "1", "volume": "165", "issue": "1", "pages": "61-74" }, { "id": "authors:m0xxa-qfp83", "collection": "authors", "collection_id": "m0xxa-qfp83", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315260", "type": "book_section", "title": "The Acquisition of Cell Fate in Mouse Development: How Do Cells First Become Heterogeneous?", "book_title": "Essays on Developmental Biology, Part B", "author": [ { "family_name": "Graham", "given_name": "Sarah J. L.", "clpid": "Graham-S-J-L" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "contributor": [ { "family_name": "Wassarman", "given_name": "Paul M.", "clpid": "Wassarman-P-M" } ], "abstract": "Specification of cell fate in the early mouse embryo involves the generation of heterogeneities between cells in polarity, position, gene expression, and protein activity. Identifying when and how these differences first become established represents an important challenge for understanding the roles of stochastic and cell history-dependent mechanisms in mammalian development.", "doi": "10.1016/bs.ctdb.2015.11.021", "isbn": "9780128013823", "publisher": "Elsevier", "place_of_publication": "New York, NY", "publication_date": "2016-01-21", "pages": "671-695" }, { "id": "authors:dz5vf-mz229", "collection": "authors", "collection_id": "dz5vf-mz229", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315456", "type": "article", "title": "Over-expression of Plk4 induces centrosome amplification, loss of primary cilia and associated tissue hyperplasia in the mouse", "author": [ { "family_name": "Coelho", "given_name": "Paula A.", "orcid": "0000-0003-0614-7575", "clpid": "Coelho-P-A" }, { "family_name": "Bury", "given_name": "Leah", "orcid": "0000-0002-6218-2837", "clpid": "Bury-L" }, { "family_name": "Shahbazi", "given_name": "Marta N.", "orcid": "0000-0002-1599-5747", "clpid": "Shahbazi-M-N" }, { "family_name": "Liakath-Ali", "given_name": "Kifayathullah", "clpid": "Liakath-Ali-K" }, { "family_name": "Tate", "given_name": "Peri H.", "clpid": "Tate-P-H" }, { "family_name": "Wormald", "given_name": "Sam", "clpid": "Wormald-S" }, { "family_name": "Hindley", "given_name": "Christopher J.", "clpid": "Hindley-C-J" }, { "family_name": "Huch", "given_name": "Meritxell", "clpid": "Huch-M" }, { "family_name": "Archer", "given_name": "Joy", "clpid": "Archer-J" }, { "family_name": "Skarnes", "given_name": "William C.", "clpid": "Skarnes-W-C" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" } ], "abstract": "To address the long-known relationship between supernumerary centrosomes and cancer, we have generated a transgenic mouse that permits inducible expression of the master regulator of centriole duplication, Polo-like-kinase-4 (Plk4). Over-expression of Plk4 from this transgene advances the onset of tumour formation that occurs in the absence of the tumour suppressor p53. Plk4 over-expression also leads to hyperproliferation of cells in the pancreas and skin that is enhanced in a p53 null background. Pancreatic islets become enlarged following Plk4 over-expression as a result of equal expansion of \u03b1- and \u03b2-cells, which exhibit centrosome amplification. Mice overexpressing Plk4 develop grey hair due to a loss of differentiated melanocytes and bald patches of skin associated with a thickening of the epidermis. This reflects an increase in proliferating cells expressing keratin 5 in the basal epidermal layer and the expansion of these cells into suprabasal layers. Such cells also express keratin 6, a marker for hyperplasia. This is paralleled by a decreased expression of later differentiation markers, involucrin, filaggrin and loricrin. Proliferating cells showed an increase in centrosome number and a loss of primary cilia, events that were mirrored in primary cultures of keratinocytes established from these animals. We discuss how repeated duplication of centrioles appears to prevent the formation of basal bodies leading to loss of primary cilia, disruption of signalling and thereby aberrant differentiation of cells within the epidermis. The absence of p53 permits cells with increased centrosomes to continue dividing, thus setting up a neoplastic state of error prone mitoses, a prerequisite for cancer development.", "doi": "10.1098/rsob.150209", "pmcid": "PMC4703062", "issn": "2046-2441", "publisher": "Royal Society", "publication": "Open Biology", "publication_date": "2015-12", "series_number": "12", "volume": "5", "issue": "12", "pages": "Art. No. 150209" }, { "id": "authors:jvwz4-byq98", "collection": "authors", "collection_id": "jvwz4-byq98", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315710", "type": "article", "title": "Development of the anterior-posterior axis is a self-organizing process in the absence of maternal cues in the mouse embryo", "author": [ { "family_name": "Bedzhov", "given_name": "Ivan", "clpid": "Bedzhov-I" }, { "family_name": "Bialecka", "given_name": "Monika", "clpid": "Bialecka-M" }, { "family_name": "Zielinska", "given_name": "Agata", "clpid": "Zielinska-A" }, { "family_name": "Kosalka", "given_name": "Joanna", "clpid": "Kosalka-J" }, { "family_name": "Antonica", "given_name": "Francesco", "orcid": "0000-0001-9359-9689", "clpid": "Antonica-F" }, { "family_name": "Thompson", "given_name": "Amelia J.", "clpid": "Thompson-A-J" }, { "family_name": "Franze", "given_name": "Kristian", "clpid": "Franze-K" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "[no abstract]", "doi": "10.1038/cr.2015.104", "pmcid": "PMC4670986", "issn": "1001-0602", "publisher": "Nature Publishing Group", "publication": "Cell Research", "publication_date": "2015-12", "series_number": "12", "volume": "25", "issue": "12", "pages": "1368-1371" }, { "id": "authors:sdgg5-s1030", "collection": "authors", "collection_id": "sdgg5-s1030", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315620", "type": "article", "title": "Mapping the journey from totipotency to lineage specification in the mouse embryo", "author": [ { "family_name": "Leung", "given_name": "Chuen Yan", "clpid": "Leung-Chuen-Yan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Understanding the past is to understand the present. Mammalian life, with all its complexity comes from a humble beginning of a single fertilized egg cell. Achieving this requires an enormous diversification of cellular function, the majority of which is generated through a series of cellular decisions during embryogenesis. The first decisions are made as the embryo prepares for implantation, a process that will require specialization of extra-embryonic lineages while preserving an embryonic one. In this mini-review, we will focus on the mouse as a mammalian model and discuss recent advances in the decision making process of the early embryo.", "doi": "10.1016/j.gde.2015.08.002", "issn": "0959-437X", "publisher": "Current Biology Ltd", "publication": "Current Opinion in Genetics and Development", "publication_date": "2015-10", "volume": "34", "pages": "71-76" }, { "id": "authors:9f5m8-s4p41", "collection": "authors", "collection_id": "9f5m8-s4p41", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315821", "type": "article", "title": "G&T-seq: parallel sequencing of single-cell genomes and transcriptomes", "author": [ { "family_name": "Macaulay", "given_name": "Iain C.", "clpid": "Macaulay-I-C" }, { "family_name": "Haerty", "given_name": "Wilfried", "clpid": "Haerty-W" }, { "family_name": "Kumar", "given_name": "Parveen", "clpid": "Kumar-P" }, { "family_name": "Li", "given_name": "Yang I.", "orcid": "0000-0002-0736-251X", "clpid": "Li-Yang-I" }, { "family_name": "Hu", "given_name": "Tim Xiaoming", "clpid": "Hu-Tim-Xiaoming" }, { "family_name": "Teng", "given_name": "Mabel J.", "clpid": "Teng-Mabel-J" }, { "family_name": "Goolam", "given_name": "Mubeen", "clpid": "Goolam-M" }, { "family_name": "Saurat", "given_name": "Nathalie", "clpid": "Saurat-N" }, { "family_name": "Coupland", "given_name": "Paul", "clpid": "Coupland-P" }, { "family_name": "Shirley", "given_name": "Lesley M.", "clpid": "Shirley-L-M" }, { "family_name": "Smith", "given_name": "Miriam", "orcid": "0000-0002-3321-1432", "clpid": "Smith-M" }, { "family_name": "Van der Aa", "given_name": "Niels", "clpid": "Van-der-Aa-N" }, { "family_name": "Banerjee", "given_name": "Ruby", "clpid": "Banerjee-R" }, { "family_name": "Ellis", "given_name": "Peter D.", "clpid": "Ellis-P-D" }, { "family_name": "Quail", "given_name": "Michael A", "clpid": "Quail-M-A" }, { "family_name": "Swerdlow", "given_name": "Harold P.", "clpid": "Swerdlow-H-P" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Livesey", "given_name": "Frederick J.", "clpid": "Livesey-F-J" }, { "family_name": "Ponting", "given_name": "Chris P.", "clpid": "Ponting-C-P" }, { "family_name": "Voet", "given_name": "Thierry", "clpid": "Voet-T" } ], "abstract": "The simultaneous sequencing of a single cell's genome and transcriptome offers a powerful means to dissect genetic variation and its effect on gene expression. Here we describe G&T-seq, a method for separating and sequencing genomic DNA and full-length mRNA from single cells. By applying G&T-seq to over 220 single cells from mice and humans, we discovered cellular properties that could not be inferred from DNA or RNA sequencing alone.", "doi": "10.1038/nmeth.3370", "issn": "1548-7091", "publisher": "Nature Publishing Group", "publication": "Nature Methods", "publication_date": "2015-06", "series_number": "6", "volume": "12", "issue": "6", "pages": "519-522" }, { "id": "authors:ezcct-45g97", "collection": "authors", "collection_id": "ezcct-45g97", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170315955", "type": "article", "title": "Cell death and morphogenesis during early mouse development: Are they interconnected?", "author": [ { "family_name": "Bedzhov", "given_name": "Ivan", "clpid": "Bedzhov-I" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Shortly after implantation the embryonic lineage transforms from a coherent ball of cells into polarized cup shaped epithelium. Recently we elucidated a previously unknown apoptosis\u2010independent morphogenic event that reorganizes the pluripotent lineage. Polarization cues from the surrounding basement membrane rearrange the epiblast into a polarized rosette\u2010like structure, where subsequently a central lumen is established. Thus, we provided a new model revising the current concept of apoptosis\u2010dependent epiblast morphogenesis. Cell death however has to be tightly regulated during embryogenesis to ensure developmental success. Here, we follow the stages of early mouse development and take a glimpse at the critical signaling and morphogenic events that determine cells destiny and reshape the embryonic lineage.", "doi": "10.1002/bies.201400147", "pmcid": "PMC4409078", "issn": "0265-9247", "publisher": "Wiley-Blackwell", "publication": "Bioessays", "publication_date": "2015-04", "series_number": "4", "volume": "37", "issue": "4", "pages": "372-378" }, { "id": "authors:awk2p-1hd96", "collection": "authors", "collection_id": "awk2p-1hd96", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316054", "type": "article", "title": "Maternal-zygotic knockout reveals a critical role of Cdx2 in the morula to blastocyst transition", "author": [ { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Cox", "given_name": "Andy", "clpid": "Cox-Andy" }, { "family_name": "Wicher", "given_name": "Krzysztof B.", "clpid": "Wicher-K-B" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The first lineage segregation in the mouse embryo generates the inner cell mass (ICM), which gives rise to the pluripotent epiblast and therefore the future embryo, and the trophectoderm (TE), which will build the placenta. The TE lineage depends on the transcription factor Cdx2. However, when Cdx2 first starts to act remains unclear. Embryos with zygotic deletion of Cdx2 develop normally until the late blastocyst stage leading to the conclusion that Cdx2 is important for the maintenance but not specification of the TE. In contrast, down-regulation of Cdx2 transcripts from the early embryo stage results in defects in TE specification before the blastocyst stage. Here, to unambiguously address at which developmental stage Cdx2 becomes first required, we genetically deleted Cdx2 from the oocyte stage using a Zp3-Cre/loxP strategy. Careful assessment of a large cohort of Cdx2 maternal-zygotic null embryos, all individually filmed, examined and genotyped, reveals an earlier lethal phenotype than observed in Cdx2 zygotic null embryos that develop until the late blastocyst stage. The developmental failure of Cdx2 maternal-zygotic null embryos is associated with cell death and failure of TE specification, starting at the morula stage. These results indicate that Cdx2 is important for the correct specification of TE from the morula stage onwards and that both maternal and zygotic pools of Cdx2 are required for correct pre-implantation embryogenesis.", "doi": "10.1016/j.ydbio.2014.12.004", "pmcid": "PMC4319684", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2015-02-15", "series_number": "2", "volume": "398", "issue": "2", "pages": "147-152" }, { "id": "authors:9427w-etp59", "collection": "authors", "collection_id": "9427w-etp59", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316127", "type": "article", "title": "BMP signalling regulates the pre-implantation development of extra-embryonic cell lineages in the mouse embryo", "author": [ { "family_name": "Graham", "given_name": "Sarah J. L.", "clpid": "Graham-S-J-L" }, { "family_name": "Wicher", "given_name": "Krzysztof B.", "clpid": "Wicher-K-B" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Guo", "given_name": "Guoji", "clpid": "Guo-Guoji" }, { "family_name": "Herath", "given_name": "Wishva", "clpid": "Herath-W" }, { "family_name": "Robson", "given_name": "Paul", "clpid": "Robson-P" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Pre-implantation development requires the specification and organization of embryonic and extra-embryonic lineages. The separation of these lineages takes place when asymmetric divisions generate inside and outside cells that differ in polarity, position and fate. Here we assess the global transcriptional identities of these precursor cells to gain insight into the molecular mechanisms regulating lineage segregation. Unexpectedly, this reveals that complementary components of the bone morphogenetic protein (BMP) signalling pathway are already differentially expressed after the first wave of asymmetric divisions. We investigate the role of BMP signalling by expressing dominant negative forms of Smad4 and Bmpr2, by downregulating the pathway using RNA interference against BMP ligands and by applying three different BMP inhibitors at distinct stages. This reveals that BMP signalling regulates the correct development of both extra-embryonic lineages, primitive endoderm and trophectoderm, but not the embryonic lineage, before implantation. Together, these findings indicate multiple roles of BMP signalling in the early mouse embryo.", "doi": "10.1038/ncomms6667", "pmcid": "PMC4338527", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2014-12-16", "volume": "5", "pages": "Art. No. 5667" }, { "id": "authors:j0dkf-09y90", "collection": "authors", "collection_id": "j0dkf-09y90", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190409-154015645", "type": "article", "title": "Developmental plasticity, cell fate specification and morphogenesis in the early mouse embryo", "author": [ { "family_name": "Bedzhov", "given_name": "Ivan", "clpid": "Bedzhov-I" }, { "family_name": "Graham", "given_name": "Sarah J. L.", "clpid": "Graham-S-J-L" }, { "family_name": "Leung", "given_name": "Chuen Yan", "clpid": "Leung-Chuen-Yan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "A critical point in mammalian development is when the early embryo implants into its mother's uterus. This event has historically been difficult to study due to the fact that it occurs within the maternal tissue and therefore is hidden from view. In this review, we discuss how the mouse embryo is prepared for implantation and the molecular mechanisms involved in directing and coordinating this crucial event. Prior to implantation, the cells of the embryo are specified as precursors of future embryonic and extra-embryonic lineages. These preimplantation cell fate decisions rely on a combination of factors including cell polarity, position and cell\u2013cell signalling and are influenced by the heterogeneity between early embryo cells. At the point of implantation, signalling events between the embryo and mother, and between the embryonic and extraembryonic compartments of the embryo itself, orchestrate a total reorganization of the embryo, coupled with a burst of cell proliferation. New developments in embryo culture and imaging techniques have recently revealed the growth and morphogenesis of the embryo at the time of implantation, leading to a new model for the blastocyst to egg cylinder transition. In this model, pluripotent cells that will give rise to the fetus self-organize into a polarized three-dimensional rosette-like structure that initiates egg cylinder formation.", "doi": "10.1098/rstb.2013.0538", "pmcid": "PMC4216461", "issn": "0962-8436", "publisher": "Royal Society of London", "publication": "Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences", "publication_date": "2014-12-05", "series_number": "1657", "volume": "369", "issue": "1657", "pages": "Art. No. 20130538" }, { "id": "authors:yk17m-8t352", "collection": "authors", "collection_id": "yk17m-8t352", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190409-162629840", "type": "article", "title": "From pluripotency to differentiation: laying foundations for the body pattern in the mouse embryo", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Hadjantonakis", "given_name": "Anna-Katerina", "clpid": "Hadjantonakis-A-K" } ], "abstract": "A central question in contemporary stem and developmental biology and modern medicine is how developmental potential becomes progressively restricted as development proceeds. How totipotency\u2014namely the ability to give rise to both embryonic and extraembryonic tissues and, in an ideal situation to the entire organism\u2014is lost. How do cells choose between alternative fates, and how do they stereotypically organize themselves into developing tissues? Today, one can strip differentiated cells of their identity by inducing pluripotency, but it is still largely unknown how pluripotency emerges in its native context within the early embryo. Indeed, the only true pluripotent cell population\u2014the epiblast\u2014exists in vivo within the early mammalian embryo.", "doi": "10.1098/rstb.2013.0535", "pmcid": "PMC4216458", "issn": "0962-8436", "publisher": "Royal Society of London", "publication": "Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences", "publication_date": "2014-12-05", "series_number": "1657", "volume": "369", "issue": "1657", "pages": "Art. No. 20130535" }, { "id": "authors:30xpv-e4042", "collection": "authors", "collection_id": "30xpv-e4042", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316220", "type": "article", "title": "In vitro culture of mouse blastocysts beyond the implantation stages", "author": [ { "family_name": "Bedzhov", "given_name": "Ivan", "clpid": "Bedzhov-I" }, { "family_name": "Leung", "given_name": "Chuen Yan", "clpid": "Leung-Chuen-Yan" }, { "family_name": "Bialecka", "given_name": "Monika", "clpid": "Bialecka-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The implanting mouse blastocyst invades the uterine stroma and undergoes a dramatic transformation into an egg cylinder. The morphogenetic and signaling events during this transition are largely unexplored, as the uterine tissues engulf the embryo. Here we describe a protocol supporting the development of the mouse embryo beyond the blastocyst stage in vitro. We established two types of medium to be applied sequentially, and we used a substrate permitting high-resolution imaging of the transition from blastocyst to egg cylinder. We developed two variants of this protocol: the first starts with intact early blastocysts that upon zona removal can attach to the substrate and develop into egg cylinders after 5 d, and the second starts with late blastocysts that upon dissection of the mural trophectoderm form egg cylinders in only 3 d. This method allows observation of a previously hidden period of development, and it provides a platform for novel research into peri-implantation embryogenesis and beyond.", "doi": "10.1038/nprot.2014.186", "issn": "1754-2189", "publisher": "Nature Publishing Group", "publication": "Nature Protocols", "publication_date": "2014-12", "series_number": "12", "volume": "9", "issue": "12", "pages": "2732-2739" }, { "id": "authors:1ga6f-c0b48", "collection": "authors", "collection_id": "1ga6f-c0b48", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316275", "type": "article", "title": "The basal position of nuclei is one pre-requisite for asymmetric cell divisions in the early mouse embryo", "author": [ { "family_name": "Ajduk", "given_name": "Anna", "clpid": "Ajduk-A" }, { "family_name": "Biswas Shivhare", "given_name": "Sourima", "orcid": "0000-0002-3704-1186", "clpid": "Biswas-Shivhare-S" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The early mouse embryo undertakes two types of cell division: symmetric that gives rise to the trophectoderm and then placenta or asymmetric that gives rise to inner cells that generate the embryo proper. Although cell division orientation is important, the mechanism regulating it has remained unclear. Here, we identify the relationship between the plane of cell division and the position of the nucleus and go towards identifying the mechanism behind it. We first find that as the 8-cell embryo progresses through the cell cycle, the nuclei of most \u2013 but not all \u2013 cells move from apical to more basal positions, in a microtubule- and kinesin-dependent manner. We then find that all asymmetric divisions happen when nuclei are located basally and, in contrast, all cells, in which nuclei remain apical, divide symmetrically. To understand the potential mechanism behind this, we determine the effects of modulating expression of Cdx2, a transcription factor key for trophectoderm formation and cell polarity. We find that increased expression of Cdx2 leads to an increase in a number of apical nuclei, whereas down-regulation of Cdx2 leads to more nuclei moving basally, which explains a previously identified relationship between Cdx2 and cell division orientation. Finally, we show that down-regulation of aPKC, involved in cell polarity, decreases the number of apical nuclei and doubles the number of asymmetric divisions. These results suggest a model in which the mutual interdependence of Cdx2 and cell polarity affects the cytoskeleton-dependent positioning of nuclei and, in consequence, the plane of cell division in the early mouse embryo.", "doi": "10.1016/j.ydbio.2014.05.009", "pmcid": "PMC4111899", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2014-08-15", "series_number": "2", "volume": "392", "issue": "2", "pages": "133-140" }, { "id": "authors:emrpc-5z719", "collection": "authors", "collection_id": "emrpc-5z719", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190409-163953574", "type": "article", "title": "A tribute to Sir John Gurdon", "author": [ { "family_name": "Rosenthal", "given_name": "Nadia", "clpid": "Rosenthal-Nadia" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Since he was awarded the Nobel Prize in 2012, John Gurdon\u05f3s remarkable contributions to biology have been celebrated in innumerable reviews and commentaries. He has since claimed that his early observations, that nuclei from differentiated cells could be reprogrammed by the factors in a fertilized egg to rerun the entire embryonic program, should not surprise us \u2013 after all, the sperm nucleus is reprogrammed by the oocyte with every fertilization event. Indeed John\u05f3s original reports in the 1970s, documenting a pluripotent stem cell state in nuclei from fully differentiated frog intestinal epithelial cells induced by the egg cytoplasm, were to wait almost forty years before receiving the accolades they deserved.", "doi": "10.1016/j.diff.2014.11.001", "issn": "0301-4681", "publisher": "Elsevier", "publication": "Differentiation", "publication_date": "2014-07", "series_number": "1", "volume": "88", "issue": "1", "pages": "1-2" }, { "id": "authors:0egyf-qvk75", "collection": "authors", "collection_id": "0egyf-qvk75", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316424", "type": "article", "title": "Citrullination regulates pluripotency and histone H1 binding to chromatin", "author": [ { "family_name": "Christophorou", "given_name": "Maria A.", "clpid": "Christophorou-M-A" }, { "family_name": "Castelo-Branco", "given_name": "Gon\u00e7alo", "clpid": "Castelo-Branco-G" }, { "family_name": "Halley-Stott", "given_name": "Richard P.", "clpid": "Halley-Stott-R-P" }, { "family_name": "Oliveira", "given_name": "Clara Slade", "clpid": "Oliveira-C-S" }, { "family_name": "Loos", "given_name": "Remco", "clpid": "Loos-R" }, { "family_name": "Radzisheuskaya", "given_name": "Aliaksandra", "clpid": "Radzisheuskaya-A" }, { "family_name": "Mowen", "given_name": "Kerri A.", "clpid": "Mowen-K-A" }, { "family_name": "Bertone", "given_name": "Paul", "clpid": "Bertone-P" }, { "family_name": "Silva", "given_name": "Jos\u00e9 C. R.", "clpid": "Silva-J-C-R" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Nielsen", "given_name": "Michael L.", "clpid": "Nielsen-M-L" }, { "family_name": "Gurdon", "given_name": "John B.", "clpid": "Gurdon-J-B" }, { "family_name": "Kouzarides", "given_name": "Tony", "clpid": "Kouzarides-T" } ], "abstract": "Citrullination is the post-translational conversion of an arginine residue within a protein to the non-coded amino acid citrulline. This modification leads to the loss of a positive charge and reduction in hydrogen-bonding ability. It is carried out by a small family of tissue-specific vertebrate enzymes called peptidylarginine deiminases (PADIs) and is associated with the development of diverse pathological states such as autoimmunity, cancer, neurodegenerative disorders, prion diseases and thrombosis. Nevertheless, the physiological functions of citrullination remain ill-defined, although citrullination of core histones has been linked to transcriptional regulation and the DNA damage response. PADI4 (also called PAD4 or PADV), the only PADI with a nuclear localization signal, was previously shown to act in myeloid cells where it mediates profound chromatin decondensation during the innate immune response to infection. Here we show that the expression and enzymatic activity of Padi4 are also induced under conditions of ground-state pluripotency and during reprogramming in mouse. Padi4 is part of the pluripotency transcriptional network, binding to regulatory elements of key stem-cell genes and activating their expression. Its inhibition lowers the percentage of pluripotent cells in the early mouse embryo and significantly reduces reprogramming efficiency. Using an unbiased proteomic approach we identify linker histone H1 variants, which are involved in the generation of compact chromatin, as novel PADI4 substrates. Citrullination of a single arginine residue within the DNA-binding site of H1 results in its displacement from chromatin and global chromatin decondensation. Together, these results uncover a role for citrullination in the regulation of pluripotency and provide new mechanistic insights into how citrullination regulates chromatin compaction.", "doi": "10.1038/nature12942", "pmcid": "PMC4843970", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2014-03-06", "series_number": "7490", "volume": "507", "issue": "7490", "pages": "104-108" }, { "id": "authors:3229t-jjj47", "collection": "authors", "collection_id": "3229t-jjj47", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316365", "type": "article", "title": "Self-Organizing Properties of Mouse Pluripotent Cells Initiate Morphogenesis upon Implantation", "author": [ { "family_name": "Bedzhov", "given_name": "Ivan", "clpid": "Bedzhov-I" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Transformation of pluripotent epiblast cells into a cup-shaped epithelium as the mouse blastocyst implants is a poorly understood and yet key developmental step. Studies of morphogenesis in embryoid bodies led to the current belief that it is programmed cell death that shapes the epiblast. However, by following embryos developing in vivo and in vitro, we demonstrate that not cell death but a previously unknown morphogenetic event transforms the amorphous epiblast into a rosette of polarized cells. This transformation requires basal membrane-stimulated integrin signaling that coordinates polarization of epiblast cells and their apical constriction, a prerequisite for lumenogenesis. We show that basal membrane function can be substituted in vitro by extracellular matrix (ECM) proteins and that ES cells can be induced to form similar polarized rosettes that initiate lumenogenesis. Together, these findings lead to a completely revised model for peri-implantation morphogenesis in which ECM triggers the self-organization of the embryo's stem cells.", "doi": "10.1016/j.cell.2014.01.023", "pmcid": "PMC3991392", "issn": "0092-8674", "publisher": "Elsevier", "publication": "Cell", "publication_date": "2014-02-27", "series_number": "5", "volume": "156", "issue": "5", "pages": "1032-1044" }, { "id": "authors:g3jpk-7dm80", "collection": "authors", "collection_id": "g3jpk-7dm80", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316524", "type": "article", "title": "Spindle Formation in the Mouse Embryo Requires Plk4 in the Absence of Centrioles", "author": [ { "family_name": "Coelho", "given_name": "Paula\u00a0A.", "orcid": "0000-0003-0614-7575", "clpid": "Coelho-P-A" }, { "family_name": "Bury", "given_name": "Leah", "orcid": "0000-0002-6218-2837", "clpid": "Bury-L" }, { "family_name": "Sharif", "given_name": "Bedra", "clpid": "Sharif-B" }, { "family_name": "Riparbelli", "given_name": "Maria\u00a0G.", "clpid": "Riparbelli-M-G" }, { "family_name": "Fu", "given_name": "Jingyan", "clpid": "Fu-Jingyan" }, { "family_name": "Callaini", "given_name": "Giuliano", "clpid": "Callaini-G" }, { "family_name": "Glover", "given_name": "David\u00a0M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "During the first five rounds of cell division in the mouse embryo, spindles assemble in the absence of centrioles. Spindle formation initiates around chromosomes, but the microtubule nucleating process remains unclear. Here we demonstrate that Plk4, a protein kinase known as a master regulator of centriole formation, is also essential for spindle assembly in the absence of centrioles. Depletion of maternal Plk4 prevents nucleation and growth of microtubules and results in monopolar spindle formation. This leads to cytokinesis failure and, consequently, developmental arrest. We show that Plk4 function depends on its kinase activity and its partner protein, Cep152. Moreover, tethering Cep152 to cellular membranes sequesters Plk4 and is sufficient to trigger spindle assembly from ectopic membranous sites. Thus, the Plk4-Cep152 complex has an unexpected role in promoting microtubule nucleation in the vicinity of chromosomes to mediate bipolar spindle formation in the absence of centrioles.", "doi": "10.1016/j.devcel.2013.09.029", "pmcid": "PMC3898710", "issn": "1534-5807", "publisher": "Cell Press", "publication": "Developmental Cell", "publication_date": "2013-12-09", "series_number": "5", "volume": "27", "issue": "5", "pages": "586-597" }, { "id": "authors:pztb2-1s788", "collection": "authors", "collection_id": "pztb2-1s788", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316664", "type": "article", "title": "The differential response to Fgf signalling in cells internalized at different times influences lineage segregation in preimplantation mouse embryos", "author": [ { "family_name": "Morris", "given_name": "Samantha A.", "clpid": "Morris-S-A" }, { "family_name": "Graham", "given_name": "Sarah J. L.", "clpid": "Graham-S-J-L" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Lineage specification in the preimplantation mouse embryo is a regulative process. Thus, it has been difficult to ascertain whether segregation of the inner-cell-mass (ICM) into precursors of the pluripotent epiblast (EPI) and the differentiating primitive endoderm (PE) is random or influenced by developmental history. Here, our results lead to a unifying model for cell fate specification in which the time of internalization and the relative contribution of ICM cells generated by two waves of asymmetric divisions influence cell fate. We show that cells generated in the second wave express higher levels of Fgfr2 than those generated in the first, leading to ICM cells with varying Fgfr2 expression. To test whether such heterogeneity is enough to bias cell fate, we upregulate Fgfr2 and show it directs cells towards PE. Our results suggest that the strength of this bias is influenced by the number of cells generated in the first wave and, mostly likely, by the level of Fgf signalling in the ICM. Differences in the developmental potential of eight-cell- and 16-cell-stage outside blastomeres placed in the inside of chimaeric embryos further support this conclusion. These results unite previous findings demonstrating the importance of developmental history and Fgf signalling in determining cell fate.", "doi": "10.1098/rsob.130104", "pmcid": "PMC3843820", "issn": "2046-2441", "publisher": "Royal Society", "publication": "Open Biology", "publication_date": "2013-11-01", "series_number": "11", "volume": "3", "issue": "11", "pages": "Art. No. 130104" }, { "id": "authors:f2a20-hky19", "collection": "authors", "collection_id": "f2a20-hky19", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316577", "type": "article", "title": "Introduction to the special issue \"Molecular Players in Early Pregnancy\"", "author": [ { "family_name": "Duan", "given_name": "En-kui", "clpid": "Duan-En-kui" }, { "family_name": "Wang", "given_name": "Haibin", "clpid": "Wang-Haibin" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "[no abstract]", "doi": "10.1016/s0098-2997(13)00054-x", "issn": "0098-2997", "publisher": "Elsevier", "publication": "Molecular Aspects of Medicine", "publication_date": "2013-10", "series_number": "5", "volume": "34", "issue": "5", "pages": "vi-vii" }, { "id": "authors:h686z-gr275", "collection": "authors", "collection_id": "h686z-gr275", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190411-124904165", "type": "article", "title": "Quality control of embryo development", "author": [ { "family_name": "Ajduk", "given_name": "Anna", "clpid": "Ajduk-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Although in recent years we have seen an undeniable improvement in the field of reproductive biology and medicine, the efficiency of in vitro fertilization (IVF) procedures remains relatively low, ranging from 4% to 40% depending on the patient's age. It is believed that this is in a large part caused by inaccurate assessment of embryo quality prior to transfer to mothers-to-be. Thus there is a strong need for further refinement of existing selection methods and development of novel, robust and, if only possible, non-invasive procedures to ensure that only embryos with the highest developmental potential are chosen for transfer. In the present review we compare various methods for assessing the quality of preimplantation embryos either currently used in IVF clinics or still to be tested. These methods include assessment of embryonic morphology, the genetic material, the transcriptomes of the oocyte and its accompanying follicular cells, and the embryo's metabolism. We discuss what information these parameters actually provide about the processes occurring in the embryo itself. We also present novel methods for selecting healthy embryos based on most recent advanced time-lapse imaging techniques, which show great promise and are likely to lead to increased in vitro fertilization efficiency.", "doi": "10.1016/j.mam.2013.03.001", "issn": "0098-2997", "publisher": "Elsevier", "publication": "Molecular Aspects of Medicine", "publication_date": "2013-10", "series_number": "5", "volume": "34", "issue": "5", "pages": "903-918" }, { "id": "authors:qs5qf-s2095", "collection": "authors", "collection_id": "qs5qf-s2095", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316805", "type": "article", "title": "Angiomotin prevents pluripotent lineage differentiation in mouse embryos via Hippo pathway-dependent and -independent mechanisms", "author": [ { "family_name": "Leung", "given_name": "Chuen Yan", "clpid": "Leung-Chuen-Yan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Cell identity is specified in the early mammalian embryo by the generation of precursors for two cell lineages: the pluripotent inner cell mass and differentiating trophectoderm. Here we identify Angiomotin as a key regulator of this process. We show that the loss of Angiomotin, together with Angiomotin-like 2, leads to differentiation of inner cell mass cells and compromised peri-implantation development. We show that Angiomotin regulates localization of Yap, and Yap-binding motifs are required for full activity of Angiomotin. Importantly, we also show that Angiomotin function can compensate for the absence of Lats1/2 kinases, indicating the ability of Angiomotin to bypass the classical Hippo pathway for Yap regulation. In polarized outside cells, Angiomotin localizes apically, pointing to the importance of cell polarity in regulating Yap to promote differentiation. We propose that both Hippo pathway-dependent and Hippo pathway-independent mechanisms regulate Yap localization to set apart pluripotent and differentiated lineages in the pre-implantation mouse embryo.", "doi": "10.1038/ncomms3251", "pmcid": "PMC3741640", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2013-08-01", "volume": "4", "pages": "Art. No. 2251" }, { "id": "authors:veh2a-a8925", "collection": "authors", "collection_id": "veh2a-a8925", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190405-170316855", "type": "article", "title": "Asymmetric Localization of Cdx2 mRNA during the First Cell-Fate Decision in Early Mouse Development", "author": [ { "family_name": "Skamagki", "given_name": "Maria", "clpid": "Skamagki-M" }, { "family_name": "Wicher", "given_name": "Krzysztof\u00a0B.", "clpid": "Wicher-K-B" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Ganguly", "given_name": "Sujoy", "clpid": "Ganguly-S" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "A longstanding question in mammalian development is whether the divisions that segregate pluripotent progenitor cells for the future embryo from cells that differentiate into extraembryonic structures are asymmetric in cell-fate instructions. The transcription factor Cdx2 plays a key role in the first cell-fate decision. Here, using live-embryo imaging, we show that localization of Cdx2 transcripts becomes asymmetric during development, preceding cell lineage segregation. Cdx2 transcripts preferentially localize apically at the late eight-cell stage and become inherited asymmetrically during divisions that set apart pluripotent and differentiating cells. Asymmetric localization depends on a cis element within the coding region of Cdx2 and requires cell polarization as well as intact microtubule and actin cytoskeletons. Failure to enrich Cdx2 transcripts apically results in a significant decrease in the number of pluripotent cells. We discuss how the asymmetric localization and segregation of Cdx2 transcripts could contribute to multiple mechanisms that establish different cell fates in the mouse embryo.", "doi": "10.1016/j.celrep.2013.01.006", "pmcid": "PMC3607255", "issn": "2211-1247", "publisher": "Elsevier", "publication": "Cell Reports", "publication_date": "2013-02-21", "series_number": "2", "volume": "3", "issue": "2", "pages": "442-457" }, { "id": "authors:5vrd6-gc482", "collection": "authors", "collection_id": "5vrd6-gc482", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190411-125134537", "type": "article", "title": "Development: Do Mouse Embryos Play Dice?", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Cells of early mouse embryo were considered for a long time to acquire cell fate at random. Recent analyses argue against this simple model.", "doi": "10.1016/j.cub.2012.10.032", "issn": "0960-9822", "publisher": "Cell Press", "publication": "Current Biology", "publication_date": "2013-01-07", "series_number": "1", "volume": "23", "issue": "1", "pages": "R15-R17" }, { "id": "authors:2928k-4zv51", "collection": "authors", "collection_id": "2928k-4zv51", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-152709124", "type": "article", "title": "Histone variant macroH2A marks embryonic differentiation in vivo and acts as an epigenetic barrier to induced pluripotency", "author": [ { "family_name": "Pasque", "given_name": "Vincent", "clpid": "Pasque-P" }, { "family_name": "Radzisheuskaya", "given_name": "Aliaksandra", "clpid": "Radzisheuskaya-A" }, { "family_name": "Gillich", "given_name": "Astrid", "clpid": "Gillich-A" }, { "family_name": "Halley-Stott", "given_name": "Richard P.", "clpid": "Halley-Stott-R-P" }, { "family_name": "Panamarova", "given_name": "Maryna", "clpid": "Panamarova-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Surani", "given_name": "M. Azim", "clpid": "Surani-M-A" }, { "family_name": "Silva", "given_name": "Jos\u00e9 C. R.", "clpid": "Silva-J-C-R" } ], "abstract": "How cell fate becomes restricted during somatic cell differentiation is a long-lasting question in biology. Epigenetic mechanisms not present in pluripotent cells and acquired during embryonic development are expected to stabilize the differentiated state of somatic cells and thereby restrict their ability to convert to another fate. The histone variant macroH2A acts as a component of an epigenetic multilayer that heritably maintains the silent X chromosome and has been shown to restrict tumor development. Here we show that macroH2A marks the differentiated cell state during mouse embryogenesis. MacroH2A.1 was found to be present at low levels upon the establishment of pluripotency in the inner cell mass and epiblast, but it was highly enriched in the trophectoderm and differentiated somatic cells later in mouse development. Chromatin immunoprecipitation revealed that macroH2A.1 is incorporated in the chromatin of regulatory regions of pluripotency genes in somatic cells such as mouse embryonic fibroblasts and adult neural stem cells, but not in embryonic stem cells. Removal of macroH2A.1, macroH2A.2 or both increased the efficiency of induced pluripotency up to 25-fold. The obtained induced pluripotent stem cells reactivated pluripotency genes, silenced retroviral transgenes and contributed to chimeras. In addition, overexpression of macroH2A isoforms prevented efficient reprogramming of epiblast stem cells to na\u00efve pluripotency. In summary, our study identifies for the first time a link between an epigenetic mark and cell fate restriction during somatic cell differentiation, which helps to maintain cell identity and antagonizes induction of a pluripotent stem cell state.", "doi": "10.1242/jcs.113019", "pmcid": "PMC3585521", "issn": "0021-9533", "publisher": "Company of Biologists", "publication": "Journal of Cell Science", "publication_date": "2012-12-15", "series_number": "24", "volume": "125", "issue": "24", "pages": "6094-6104" }, { "id": "authors:r7qbs-xnm48", "collection": "authors", "collection_id": "r7qbs-xnm48", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-152709752", "type": "article", "title": "Developmental Plasticity Is Bound by Pluripotency and the Fgf and Wnt Signaling Pathways", "author": [ { "family_name": "Morris", "given_name": "Samantha\u00a0A.", "clpid": "Morris-S-A" }, { "family_name": "Guo", "given_name": "Yu", "clpid": "Guo-Yu" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Plasticity is a well-known feature of mammalian development, and yet very little is known about its underlying mechanism. Here, we establish a model system to examine the extent and limitations of developmental plasticity in living mouse embryos. We show that halved embryos follow the same strict clock of developmental transitions as intact embryos, but their potential is not equal. We have determined that unless a minimum of four pluripotent cells is established before implantation, development will arrest. This failure can be rescued by modulating Fgf and Wnt signaling to enhance pluripotent cell number, allowing the generation of monozygotic twins, which is an otherwise rare phenomenon. Knowledge of the minimum pluripotent-cell number required for development to birth, as well as the different potentials of blastomeres, allowed us to establish a protocol for splitting an embryo into one part that develops to adulthood and another that provides embryonic stem cells for that individual.", "doi": "10.1016/j.celrep.2012.08.029", "pmcid": "PMC3607220", "issn": "2211-1247", "publisher": "Elsevier", "publication": "Cell Reports", "publication_date": "2012-10-25", "series_number": "4", "volume": "2", "issue": "4", "pages": "756-765" }, { "id": "authors:ry6xx-fyb59", "collection": "authors", "collection_id": "ry6xx-fyb59", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-152709819", "type": "article", "title": "Protein Arginine Methyltransferase 6 Regulates Embryonic Stem Cell Identity", "author": [ { "family_name": "Lee", "given_name": "Yun Hwa", "clpid": "Lee-Yun-Hwa" }, { "family_name": "Ma", "given_name": "Hui", "clpid": "Ma-Hui" }, { "family_name": "Tan", "given_name": "Tuan Zea", "clpid": "Tan-Tuan-Zea" }, { "family_name": "Ng", "given_name": "Swee Siang", "clpid": "Ng-Swee-Siang" }, { "family_name": "Soong", "given_name": "Richie", "clpid": "Soong-Richie" }, { "family_name": "Mori", "given_name": "Seiichi", "clpid": "Mori-Seiichi" }, { "family_name": "Fu", "given_name": "Xin-Yuan", "clpid": "Fu-Xin-Yuan" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Wu", "given_name": "Qiang", "clpid": "Wu-Qiang" } ], "abstract": "Histone arginine methylation has emerged as an important histone modification involved in gene regulation. Protein arginine methyltransferase (PRMT) 4 and 5 have been shown to play essential roles in early embryonic development and in embryonic stem (ES) cells. Recently, it has been reported that PRMT6-mediated di-methylation of histone H3 at arginine 2 (H3R2me2) can antagonize tri-methylation of histone H3 at lysine 4 (H3K4me3), which marks active genes. However, whether PRMT6 and PRMT6-mediated H3R2me2 play crucial roles in early embryonic development and ES cell identity remain unclear. Here, we have investigated their roles using gain and loss of function studies with mouse ES cells as a model system. We report that Prmt6 and histone H3R2 methylation levels increased when ES cells are induced to differentiate. Consistently, we find that differentiation of ES cells upon upregulation of Prmt6 is associated with decreased expression of pluripotency genes and increased expression of differentiation markers. We also observe that elevation of Prmt6 increases the methylation level of histone H3R2 and decreases H3K4me, Chd1, and Wdr5 levels at the promoter regions of Oct4 and Nanog. Surprisingly, knockdown of Prmt6 also leads to downregulation of pluripotency genes and induction of expression of differentiation markers suggesting that Prmt6 is important for ES cell pluripotency and self-renewal. Our results indicate that a critical level of Prmt6 and histone H3R2me must be maintained in mouse ES cells to sustain their pluripotency.", "doi": "10.1089/scd.2011.0330", "pmcid": "PMC5729635", "issn": "1547-3287", "publisher": "Mary Ann Liebert", "publication": "Stem Cells and Development", "publication_date": "2012-09-20", "series_number": "14", "volume": "21", "issue": "14", "pages": "2613-2622" }, { "id": "authors:a03mj-0y409", "collection": "authors", "collection_id": "a03mj-0y409", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-152710198", "type": "book_section", "title": "Formation of Distinct Cell Types in the Mouse Blastocyst", "book_title": "Mouse Development: From Oocyte to Stem Cells", "author": [ { "family_name": "Morris", "given_name": "Samantha A.", "clpid": "Morris-S-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "contributor": [ { "family_name": "Kubiak", "given_name": "Jacek Z.", "clpid": "Kubiak-J-Z" } ], "abstract": "Early development of the mouse comprises a sequence of cell fate decisions in which cells are guided along a pathway of restricted potential and increasing specialisation. The first choice faced by cells of the embryo is whether to become trophectoderm (TE) or inner cell mass (ICM); TE is an extra-embryonic tissue which will form the embryonic portion of the placenta, whilst ICM gives rise to cells responsible for generating the foetus. In the second cell fate decision, the ICM is further refined into pluripotent cells forming the future body of the embryo, epiblast (EPI) and extra-embryonic primitive endoderm (PE), a tissue essential for patterning the embryo and establishing the developmental circulation. Understanding this early lineage segregation is critical for informing attempts to capture pluripotency and direct cell fate in vitro. Unlike the predictability of nonmammalian cell fate, development of the mouse embryo retains the flexibility to adapt to changing circumstances during development. Here we describe these first cell fate decisions, how they can be biased whilst maintaining flexibility and, finally, some of the molecular circuitry underlying early fate choice.", "doi": "10.1007/978-3-642-30406-4_11", "isbn": "9783642304057", "publisher": "Springer Berlin Heidelberg", "place_of_publication": "Berlin, Heidelberg", "publication_date": "2012-07-21", "pages": "203-217" }, { "id": "authors:tkrsr-rsa52", "collection": "authors", "collection_id": "tkrsr-rsa52", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-152710123", "type": "article", "title": "Advances in embryo selection methods", "author": [ { "family_name": "Ajduk", "given_name": "Anna", "clpid": "Ajduk-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Despite many recent advances in the field of reproductive biology and medicine, the efficiency of in vitro fertilization procedures remains relatively low. There is a need for a reliable and non-invasive method of embryo selection to ensure that only embryos with the highest developmental potential are chosen for transfer to mothers-to-be. Here, we compare various methods currently used for assessing embryonic viability, such as examination of embryonic morphology, quality of the genetic material, or metabolism. Additionally, we discuss novel procedures for embryonic assessment based on advanced time-lapse imaging techniques, which show great promise and may lead to increased in vitro fertilization efficiencies.", "doi": "10.3410/b4-11", "pmcid": "PMC3369238", "issn": "1757-594X", "publisher": "Faculty of 1000", "publication": "F1000 Biology Reports", "publication_date": "2012-06-01", "volume": "4", "pages": "Art. No. 11" }, { "id": "authors:4z299-8kg52", "collection": "authors", "collection_id": "4z299-8kg52", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-152709920", "type": "article", "title": "Phospholipase C-\u03b6-induced Ca^(2+) oscillations cause coincident cytoplasmic movements in human oocytes that failed to fertilize after intracytoplasmic sperm injection", "author": [ { "family_name": "Swann", "given_name": "Karl", "clpid": "Swann-K" }, { "family_name": "Windsor", "given_name": "Shane", "clpid": "Windsor-S" }, { "family_name": "Campbell", "given_name": "Karen", "clpid": "Campbell-K" }, { "family_name": "Elgmati", "given_name": "Khalil", "clpid": "Elgmati-K" }, { "family_name": "Nomikos", "given_name": "Michail", "clpid": "Nomikos-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Amso", "given_name": "Nazar", "clpid": "Amso-N" }, { "family_name": "Lai", "given_name": "F. Anthony", "clpid": "Lai-F-Anthony" }, { "family_name": "Thomas", "given_name": "Adrian", "clpid": "Thomas-A" }, { "family_name": "Graham", "given_name": "Christopher", "clpid": "Graham-C" } ], "abstract": "OBJECTIVE: To evaluate the imaging of cytoplasmic movements in human oocytes as a potential method to monitor the pattern of Ca^(2+) oscillations during activation. \n\nDESIGN: Test of a laboratory technique. \n\nSETTING: University medical school research laboratory. \n\nPATIENT(S): Donated unfertilized human oocytes from intracytoplasmic sperm injection (ICSI) cycles. \n\nINTERVENTION(S): Microinjection of oocytes with phospholipase C (PLC) zeta (\u03b6) cRNA and a Ca^(2+)-sensitive fluorescent dye. \n\nMAIN OUTCOME MEASURE(S): Simultaneous detection of oocyte cytoplasmic movements using particle image velocimetry (PIV) and of Ca^(2+) oscillations using a Ca^(2+)-sensitive fluorescent dye. \n\nRESULT(S): Microinjection of PLC\u03b6 cRNA into human oocytes that had failed to fertilize after ICSI resulted in the appearance of prolonged Ca^(2+) oscillations. Each transient Ca^(2+) concentration change was accompanied by a small coordinated movement of the cytoplasm that could be detected using PIV analysis. \n\nCONCLUSION(S): The occurrence and frequency of cytoplasmic Ca^(2+) oscillations, a critical parameter in activating human zygotes, can be monitored by PIV analysis of cytoplasmic movements. This simple method provides a novel, noninvasive approach to determine in real time the occurrence and frequency of Ca^(2+) oscillations in human zygotes.", "doi": "10.1016/j.fertnstert.2011.12.013", "pmcid": "PMC3334266", "issn": "0015-0282", "publisher": "Elsevier", "publication": "Fertility and Sterility", "publication_date": "2012-03", "series_number": "3", "volume": "97", "issue": "3", "pages": "742-747" }, { "id": "authors:nwxqr-4zp41", "collection": "authors", "collection_id": "nwxqr-4zp41", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-152710020", "type": "article", "title": "Dynamics of anterior\u2013posterior axis formation in the developing mouse embryo", "author": [ { "family_name": "Morris", "given_name": "Samantha A.", "clpid": "Morris-S-A" }, { "family_name": "Grewal", "given_name": "Seema", "clpid": "Grewal-S" }, { "family_name": "Barrios", "given_name": "Florencia", "clpid": "Barrios-F" }, { "family_name": "Patankar", "given_name": "Sameer N.", "clpid": "Patankar-S-N" }, { "family_name": "Strauss", "given_name": "Bernhard", "clpid": "Strauss-B" }, { "family_name": "Buttery", "given_name": "Lee", "clpid": "Buttery-L" }, { "family_name": "Alexander", "given_name": "Morgan", "clpid": "Alexander-M" }, { "family_name": "Shakesheff", "given_name": "Kevin M.", "clpid": "Shakesheff-K-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The development of an anterior\u2013posterior (AP) polarity is a crucial process that in the mouse has been very difficult to analyse, because it takes place as the embryo implants within the mother. To overcome this obstacle, we have established an in-vitro culture system that allows us to follow the step-wise development of anterior visceral endoderm (AVE), critical for establishing AP polarity. Here we use this system to show that the AVE originates in the implanting blastocyst, but that additional cells subsequently acquire AVE characteristics. These 'older' and 'younger' AVE domains coalesce as the egg cylinder emerges from the blastocyst structure. Importantly, we show that AVE migration is led by cells expressing the highest levels of AVE marker, highlighting that asymmetry within the AVE domain dictates the direction of its migration. Ablation of such leading cells prevents AVE migration, suggesting that these cells are important for correct establishment of the AP axis.", "doi": "10.1038/ncomms1671", "pmcid": "PMC3293425", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2012-02-14", "volume": "3", "pages": "Art. No. 673" }, { "id": "authors:myztn-z9y36", "collection": "authors", "collection_id": "myztn-z9y36", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163114215", "type": "article", "title": "Rhythmic actomyosin-driven contractions induced by sperm entry predict mammalian embryo viability", "author": [ { "family_name": "Ajduk", "given_name": "Anna", "clpid": "Ajduk-A" }, { "family_name": "Ilozue", "given_name": "Tagbo", "clpid": "Ilozue-Tagbo" }, { "family_name": "Windsor", "given_name": "Shane", "clpid": "Windsor-S" }, { "family_name": "Yu", "given_name": "Yuansong", "clpid": "Yu-Yuansong" }, { "family_name": "Seres", "given_name": "K. Bianka", "clpid": "Seres-K-B" }, { "family_name": "Bomphrey", "given_name": "Richard J.", "clpid": "Bomphrey-R-J" }, { "family_name": "Tom", "given_name": "Brian D.", "clpid": "Tom-B-D" }, { "family_name": "Swann", "given_name": "Karl", "clpid": "Swann-K" }, { "family_name": "Thomas", "given_name": "Adrian", "clpid": "Thomas-A" }, { "family_name": "Graham", "given_name": "Chris", "clpid": "Graham-C" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Fertilization-induced cytoplasmic flows are a conserved feature of eggs in many species. However, until now the importance of cytoplasmic flows for the development of mammalian embryos has been unknown. Here, by combining a rapid imaging of the freshly fertilized mouse egg with advanced image analysis based on particle image velocimetry, we show that fertilization induces rhythmical cytoplasmic movements that coincide with pulsations of the protrusion forming above the sperm head. We find that these movements are caused by contractions of the actomyosin cytoskeleton triggered by Ca^(2+) oscillations induced by fertilization. Most importantly, the relationship between the movements and the events of egg activation makes it possible to use the movements alone to predict developmental potential of the zygote. In conclusion, this method offers, thus far, the earliest and fastest, non-invasive way to predict the viability of eggs fertilized in vitro and therefore can potentially improve greatly the prospects for IVF treatment.", "doi": "10.1038/ncomms1424", "pmcid": "PMC3265380", "issn": "2041-1723", "publisher": "Nature Publishing Group", "publication": "Nature Communications", "publication_date": "2011-08-09", "volume": "2", "pages": "Art. No. 417" }, { "id": "authors:3pv6s-bsg43", "collection": "authors", "collection_id": "3pv6s-bsg43", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163114357", "type": "article", "title": "Proclaiming fate in the early mouse embryo", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "In the mouse embryo, the first differences between cells that result in distinct lineages have long been thought to arise only as a consequence of differential cell positioning at relatively late preimplantation stages. Differences in Oct4 transcription factor kinetics between cells at the 4\u20138-cell stage are now shown to be predictive of future lineages, providing further evidence for much earlier initiation of cell fate decisions.", "doi": "10.1038/ncb0211-112", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2011-02", "series_number": "2", "volume": "13", "issue": "2", "pages": "112-114" }, { "id": "authors:10j6w-4we11", "collection": "authors", "collection_id": "10j6w-4we11", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163114460", "type": "article", "title": "The chromosome passenger complex is required for fidelity of chromosome transmission and cytokinesis in meiosis of mouse oocytes", "author": [ { "family_name": "Sharif", "given_name": "Bedra", "clpid": "Sharif-B" }, { "family_name": "Na", "given_name": "Jie", "clpid": "Na-Jie" }, { "family_name": "Lykke-Hartmann", "given_name": "Karin", "clpid": "Lykke-Hartmann-K" }, { "family_name": "McLaughlin", "given_name": "Stephen H.", "clpid": "McLaughlin-S-H" }, { "family_name": "Laue", "given_name": "Ernest", "clpid": "Laue-E" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The existence of two forms of the chromosome passenger complex (CPC) in the mammalian oocyte has meant that its role in female meiosis has remained unclear. Here we use loss- and gain-of function approaches to assess the meiotic functions of one of the shared components of these complexes, INCENP, and of the variable kinase subunits, Aurora B or Aurora C. We show that either the depletion of INCENP or the combined inhibition of Aurora kinases B and C activates the anaphase-promoting complex or cyclosome (APC/C) before chromosomes have properly congressed in meiosis I and also prevents cytokinesis and hence extrusion of the first polar body. Overexpression of Aurora C also advances APC/C activation and results in cytokinesis failure in a high proportion of oocytes, indicative of a dominant effect on CPC function. Together, this points to roles for the meiotic CPC in functions similar to the mitotic roles of the complex: correcting chromosome attachment to microtubules, facilitating the spindle-assembly checkpoint (SAC) function and enabling cytokinesis. Surprisingly, overexpression of Aurora B leads to a failure of APC/C activation, stabilization of securin and consequently a failure of chiasmate chromosomes to resolve \u2013 a dominant phenotype that is completely suppressed by depletion of INCENP. Taken together with the differential distribution of Aurora proteins B and C on chiasmate chromosomes, this points to differential functions of the two forms of CPC in regulating the separation of homologous chromosomes in meiosis I.", "doi": "10.1242/jcs.067447", "pmcid": "PMC2995614", "issn": "0021-9533", "publisher": "Company of Biologists", "publication": "Journal of Cell Science", "publication_date": "2010-12-15", "series_number": "24", "volume": "123", "issue": "24", "pages": "4292-4300" }, { "id": "authors:hkcrp-dhx49", "collection": "authors", "collection_id": "hkcrp-dhx49", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163114563", "type": "article", "title": "Stochasticity versus determinism in development: a false dichotomy?", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Huang", "given_name": "Sui", "clpid": "Huang-Sui" } ], "abstract": "Gene expression noise can suggest a role for stochasticity during development. But Magdalena Zernicka-Goetz and Sui Huang argue that care should be taken in data interpretation and language use before labelling developmental events as 'random'.", "doi": "10.1038/nrg2886", "issn": "1471-0056", "publisher": "Nature Publishing Group", "publication": "Nature Reviews. Genetics", "publication_date": "2010-11", "series_number": "11", "volume": "11", "issue": "11", "pages": "743-744" }, { "id": "authors:36cww-sj046", "collection": "authors", "collection_id": "36cww-sj046", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163114658", "type": "article", "title": "Developmental control of the early mammalian embryo: competition among heterogeneous cells that biases cell fate", "author": [ { "family_name": "Bruce", "given_name": "Alexander W.", "clpid": "Bruce-A-W" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The temporal and spatial segregation of the two extra-embryonic cell lineages, trophectoderm and primitive endoderm (TE and PE respectively), from the pluripotent epiblast (EPI) during mammalian pre-implantation development are prerequisites for the successful implantation of the blastocyst. The mechanisms underlying these earliest stages of development remain a fertile topic for research and informed debate. In recent years novel roles for various transcription factors, polarity factors and signalling cascades have been uncovered. This mini-review seeks to summarise some of this work and to put it into the context of the regulative nature of early mammalian development and to highlight how the increasing evidence of naturally occurring asymmetries and heterogeneity in the embryo can bias specification of the distinct cell types of the blastocyst.", "doi": "10.1016/j.gde.2010.05.006", "issn": "0959-437X", "publisher": "Current Biology Ltd", "publication": "Current Opinion in Genetics and Development", "publication_date": "2010-10", "series_number": "5", "volume": "20", "issue": "5", "pages": "485-491" }, { "id": "authors:g9esx-7g593", "collection": "authors", "collection_id": "g9esx-7g593", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163114865", "type": "article", "title": "Epigenetic Modification Affecting Expression of Cell Polarity and Cell Fate Genes to Regulate Lineage Specification in the Early Mouse Embryo", "author": [ { "family_name": "Parfitt", "given_name": "David-Emlyn", "clpid": "Parfitt-D-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Formation of inner and outer cells of the mouse embryo distinguishes pluripotent inner cell mass (ICM) from differentiating trophectoderm (TE). Carm1, which methylates histone H3R17 and R26, directs cells to ICM rather that TE. To understand the mechanism by which this epigenetic modification directs cell fate, we generated embryos with in vivo\u2013labeled cells of different Carm1 levels, using time-lapse imaging to reveal dynamics of their behavior, and related this to cell polarization. This shows that Carm1 affects cell fate by promoting asymmetric divisions, that direct one daughter cell inside, and cell engulfment, where neighboring cells with lower Carm1 levels compete for outside positions. This is associated with changes to the expression pattern and spatial distribution of cell polarity proteins: Cells with higher Carm1 levels show reduced expression and apical localization of Par3 and a dramatic increase in expression of PKCII, antagonist of the apical protein aPKC. Expression and basolateral localization of the mouse Par1 homologue, EMK1, increases concomitantly. Increased Carm1 also reduces Cdx2 expression, a transcription factor key for TE differentiation. These results demonstrate how the extent of a specific epigenetic modification could affect expression of cell polarity and fate-determining genes to ensure lineage allocation in the mouse embryo.", "doi": "10.1091/mbc.e10-01-0053", "pmcid": "PMC2912351", "issn": "1059-1524", "publisher": "American Society for Cell Biology", "publication": "Molecular Biology of the Cell", "publication_date": "2010-08-01", "series_number": "15", "volume": "21", "issue": "15", "pages": "2649-2660" }, { "id": "authors:gz5r3-g0g12", "collection": "authors", "collection_id": "gz5r3-g0g12", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163114755", "type": "article", "title": "Maternally and zygotically provided Cdx2 have novel and critical roles for early development of the mouse embryo", "author": [ { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Bruce", "given_name": "Alexander W.", "clpid": "Bruce-A-W" }, { "family_name": "Tan", "given_name": "Meng H.", "clpid": "Tan-Meng-H" }, { "family_name": "Leong", "given_name": "Denise E.", "clpid": "Leong-Denise-E" }, { "family_name": "Skamagki", "given_name": "Maria", "clpid": "Skamagki-M" }, { "family_name": "Yao", "given_name": "Mylene", "clpid": "Yao-Mylene" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Divisions of polarised blastomeres that allocate polar cells to outer and apolar cells to inner positions initiate the first cell fate decision in the mouse embryo. Subsequently, outer cells differentiate into trophectoderm while inner cells retain pluripotency to become inner cell mass (ICM) of the blastocyst. Elimination of zygotic expression of trophectoderm-specific transcription factor Cdx2 leads to defects in the maintenance of the blastocyst cavity, suggesting that it participates only in the late stage of trophectoderm formation. However, we now find that mouse embryos also have a maternally provided pool of Cdx2 mRNA. Moreover, depletion of both maternal and zygotic Cdx2 from immediately after fertilization by three independent approaches, dsRNAi, siRNAi and morpholino oligonucleotides, leads to developmental arrest at much earlier stages than expected from elimination of only zygotic Cdx2. This developmental arrest is associated with defects in cell polarisation, reflected by expression and localisation of cell polarity molecules such as Par3 and aPKC and cell compaction at the 8- and 16-cell stages. Cells deprived of Cdx2 show delayed development with increased cell cycle length, irregular cell division and increased incidence of apoptosis. Although some Cdx2-depleted embryos initiate cavitation, the cavity cannot be maintained. Furthermore, expression of trophectoderm-specific genes, Gata3 and Eomes, and also the trophectoderm-specific cytokeratin intermediate filament, recognised by Troma1, are greatly reduced or undetectable. Taken together, our results indicate that Cdx2 participates in two steps leading to trophectoderm specification: appropriate polarisation of blastomeres at the 8- and 16-cell stage and then the maintenance of trophectoderm lineage-specific differentiation.", "doi": "10.1016/j.ydbio.2010.04.017", "pmcid": "PMC2954319", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2010-08-01", "series_number": "1", "volume": "344", "issue": "1", "pages": "66-78" }, { "id": "authors:dz1h0-dr993", "collection": "authors", "collection_id": "dz1h0-dr993", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163114958", "type": "article", "title": "Origin and formation of the first two distinct cell types of the inner cell mass in the mouse embryo", "author": [ { "family_name": "Morris", "given_name": "Samantha A.", "clpid": "Morris-S-A" }, { "family_name": "Teo", "given_name": "Roy T. Y.", "clpid": "Teo-Roy-T-Y" }, { "family_name": "Li", "given_name": "Huiliang", "clpid": "Li-Huiliang" }, { "family_name": "Robson", "given_name": "Paul", "clpid": "Robson-P" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "A crucial question in mammalian development is how cells of the early embryo differentiate into distinct cell types. The first decision is taken when cells undertake waves of asymmetric division that generate one daughter on the inside and one on the outside of the embryo. After this division, some cells on the inside remain pluripotent and give rise to the epiblast, and hence the future body, whereas others develop into the primitive endoderm, an extraembryonic tissue. How the fate of these inside cells is decided is unknown: Is the process random, or is it related to their developmental origins? To address this question, we traced all cells by live-cell imaging in intact, unmanipulated embryos until the epiblast and primitive endoderm became distinct. This analysis revealed that inner cell mass (ICM) cells have unrestricted developmental potential. However, cells internalized by the first wave of asymmetric divisions are biased toward forming pluripotent epiblast, whereas cells internalized in the next two waves of divisions are strongly biased toward forming primitive endoderm. Moreover, we show that cells internalized by the second wave up-regulate expression of Gata6 and Sox17, and changing the expression of these genes determines whether the cells become primitive endoderm. Finally, with our ability to determine the origin of cells, we find that inside cells that are mispositioned when they are born can sort into the correct layer. In conclusion, we propose a model in which the timing of cell internalization, cell position, and cell sorting combine to determine distinct lineages of the preimplantation mouse embryo.", "doi": "10.1073/pnas.0915063107", "pmcid": "PMC2852013", "issn": "0027-8424", "publisher": "National Academy of Sciences", "publication": "Proceedings of the National Academy of Sciences of the United States of America", "publication_date": "2010-04-06", "series_number": "14", "volume": "107", "issue": "14", "pages": "6364-6369" }, { "id": "authors:m1a28-h5z19", "collection": "authors", "collection_id": "m1a28-h5z19", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163115057", "type": "article", "title": "CARM1 Is Required in Embryonic Stem Cells to Maintain Pluripotency and Resist Differentiation", "author": [ { "family_name": "Wu", "given_name": "Qiang", "clpid": "Wu-Qiang" }, { "family_name": "Bruce", "given_name": "Alexander W.", "clpid": "Bruce-A-W" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Ellis", "given_name": "Peter D.", "clpid": "Ellis-P-D" }, { "family_name": "Andrews", "given_name": "Robert M.", "clpid": "Andrews-R-M" }, { "family_name": "Langford", "given_name": "Cordelia F.", "clpid": "Langford-C-F" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Histone H3 methylation at R17 and R26 recently emerged as a novel epigenetic mechanism regulating pluripotency in mouse embryos. Blastomeres of four\u2010cell embryos with high H3 methylation at these sites show unrestricted potential, whereas those with lower levels cannot support development when aggregated in chimeras of like cells. Increasing histone H3 methylation, through expression of coactivator\u2010associated\u2010protein\u2010arginine\u2010methyltransferase 1 (CARM1) in embryos, elevates expression of key pluripotency genes and directs cells to the pluripotent inner cell mass. We demonstrate CARM1 is also required for the self\u2010renewal and pluripotency of embryonic stem (ES) cells. In ES cells, CARM1 depletion downregulates pluripotency genes leading to their differentiation. CARM1 associates with Oct4/Pou5f1 and Sox2 promoters that display detectable levels of R17/26 histone H3 methylation. In CARM1 overexpressing ES cells, histone H3 arginine methylation is also at the Nanog promoter to which CARM1 now associates. Such cells express Nanog at elevated levels and delay their response to differentiation signals. Thus, like in four\u2010cell embryo blastomeres, histone H3 arginine methylation by CARM1 in ES cells allows epigenetic modulation of pluripotency.", "doi": "10.1002/stem.131", "pmcid": "PMC4135545", "issn": "1066-5099", "publisher": "Wiley", "publication": "Stem Cells", "publication_date": "2009-11", "series_number": "11", "volume": "27", "issue": "11", "pages": "2637-2645" }, { "id": "authors:bq75p-kym96", "collection": "authors", "collection_id": "bq75p-kym96", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190417-163115154", "type": "article", "title": "Active cell movements coupled to positional induction are involved in lineage segregation in the mouse blastocyst", "author": [ { "family_name": "Meilhac", "given_name": "Sigol\u00e8ne M.", "clpid": "Meilhac-S-M" }, { "family_name": "Adams", "given_name": "Richard J.", "clpid": "Adams-R-J" }, { "family_name": "Morris", "given_name": "Samantha A.", "clpid": "Morris-S-A" }, { "family_name": "Danckaert", "given_name": "Anne", "clpid": "Danckaert-A" }, { "family_name": "Le Garrec", "given_name": "Jean-Fran\u00e7ois", "clpid": "Le-Garrec-J-F" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "In the mouse blastocyst, some cells of the inner cell mass (ICM) develop into primitive endoderm (PE) at the surface, while deeper cells form the epiblast. It remained unclear whether the position of cells determines their fate, such that gene expression is adjusted to cell position, or if cells are pre-specified at random positions and then sort. We have tracked and characterised dynamics of all ICM cells from the early to late blastocyst stage. Time-lapse microscopy in H2B-EGFP embryos shows that a large proportion of ICM cells change position between the surface and deeper compartments. Most of this cell movement depends on actin and is associated with cell protrusions. We also find that while most cells are precursors for only one lineage, some give rise to both, indicating that lineage segregation is not complete in the early ICM. Finally, changing the expression levels of the PE marker Gata6 reveals that it is required in surface cells but not sufficient for the re-positioning of deeper cells. We provide evidence that Wnt9A, known to be expressed in the surface ICM, facilitates re-positioning of Gata6-expressing cells. Combining these experimental results with computer modelling suggests that PE formation involves both cell sorting movements and position-dependent induction.", "doi": "10.1016/j.ydbio.2009.04.036", "pmcid": "PMC3353123", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2009-07-15", "series_number": "2", "volume": "331", "issue": "2", "pages": "210-221" }, { "id": "authors:cm1wa-4j157", "collection": "authors", "collection_id": "cm1wa-4j157", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150055208", "type": "article", "title": "Making a firm decision: multifaceted regulation of cell fate in the early mouse embryo", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Morris", "given_name": "Samantha A.", "clpid": "Morris-S-A" }, { "family_name": "Bruce", "given_name": "Alexander W.", "clpid": "Bruce-A-W" } ], "abstract": "The preimplantation mammalian embryo offers a striking opportunity to address the question of how and why apparently identical cells take on separate fates. Two cell fate decisions are taken before the embryo implants; these decisions set apart a group of pluripotent cells, progenitors for the future body, from the distinct extraembryonic lineages of trophectoderm and primitive endoderm. New molecular, cellular and developmental insights reveal the interplay of transcriptional regulation, epigenetic modifications, cell position and cell polarity in these two fate decisions in the mouse. We discuss how mechanisms proposed in previously distinct models might work in concert to progressively reinforce cell fate decisions through feedback loops.", "doi": "10.1038/nrg2564", "issn": "1471-0056", "publisher": "Nature Publishing Group", "publication": "Nature Reviews. Genetics", "publication_date": "2009-07", "series_number": "7", "volume": "10", "issue": "7", "pages": "467-477" }, { "id": "authors:s7kyp-rnd93", "collection": "authors", "collection_id": "s7kyp-rnd93", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150055370", "type": "article", "title": "Role of Cdx2 and cell polarity in cell allocation and specification of trophectoderm and inner cell mass in the mouse embryo", "author": [ { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Parfitt", "given_name": "David-Emlyn", "clpid": "Parfitt-D-E" }, { "family_name": "Guo", "given_name": "Guoji", "clpid": "Guo-Guoji" }, { "family_name": "Skamagki", "given_name": "Maria", "clpid": "Skamagki-M" }, { "family_name": "Grabarek", "given_name": "Joanna B.", "clpid": "Grabarek-J-B" }, { "family_name": "Johnson", "given_name": "Martin H.", "clpid": "Johnson-M-H" }, { "family_name": "Robson", "given_name": "Paul", "clpid": "Robson-P" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Genesis of the trophectoderm and inner cell mass (ICM) lineages occurs in two stages. It is initiated via asymmetric divisions of eight- and 16-cell blastomeres that allocate cells to inner and outer positions, each with different developmental fates. Outside cells become committed to the trophectoderm at the blastocyst stage through Cdx2 activity, but here we show that Cdx2 can also act earlier to influence cell allocation. Increasing Cdx2 levels in individual blastomeres promotes symmetric divisions, thereby allocating more cells to the trophectoderm, whereas reducing Cdx2 promotes asymmetric divisions and consequently contribution to the ICM. Furthermore, both Cdx2 mRNA and protein levels are heterogeneous at the eight-cell stage. This heterogeneity depends on cell origin and has developmental consequences. Cdx2 expression is minimal in cells with unrestricted developmental potential that contribute preferentially to the ICM and is maximal in cells with reduced potential that contribute more to the trophectoderm. Finally, we describe a mutually reinforcing relationship between cellular polarity and Cdx2: Cdx2 influences cell polarity by up-regulating aPKC, but cell polarity also influences Cdx2 through asymmetric distribution of Cdx2 mRNA in polarized blastomeres. Thus, divisions generating inside and outside cells are truly asymmetric with respect to cell fate instructions. These two interacting effects ensure the generation of a stable outer epithelium by the blastocyst stage.", "doi": "10.1101/gad.486108", "pmcid": "PMC2559904", "issn": "0890-9369", "publisher": "Cold Spring Harbor Laboratory Press", "publication": "Genes and Development", "publication_date": "2008-10-01", "series_number": "19", "volume": "22", "issue": "19", "pages": "2692-2706" }, { "id": "authors:4z8mp-mr447", "collection": "authors", "collection_id": "4z8mp-mr447", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150055486", "type": "article", "title": "Maternal Argonaute 2 Is Essential for Early Mouse Development at the Maternal-Zygotic Transition", "author": [ { "family_name": "Lykke-Andersen", "given_name": "Karin", "clpid": "Lykke-Andersen-K" }, { "family_name": "Gilchrist", "given_name": "Michael J.", "clpid": "Gilchrist-M-J" }, { "family_name": "Grabarek", "given_name": "Joanna B.", "clpid": "Grabarek-J-B" }, { "family_name": "Das", "given_name": "Partha", "clpid": "Das-P" }, { "family_name": "Miska", "given_name": "Eric", "clpid": "Miska-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Activation of zygotic gene expression in the two-cell mouse embryo is associated with destruction of maternally inherited transcripts, an important process for embryogenesis about which little is understood. We asked whether the Argonaute (Ago)/RNA-induced silencing complex, providing the mRNA \"slicer\" activity in gene silencing, might contribute to this process. Here we show that Ago2, 3, and 4 transcripts are contributed to the embryo maternally. By systematic knockdown of maternal Ago2, 3, and 4, individually and in combination, we find that only Ago2 is required for development beyond the two-cell stage. Knockdown of Ago2 stabilizes one set of maternal mRNAs and reduces zygotic transcripts of another set of genes. Ago2 is localized in mRNA-degradation P-bodies analogous to those that function in RNAi-like mechanisms in other systems. Profiling the expression of microRNAs throughout preimplantation development identified several candidates that could potentially work with Ago2 to mediate degradation of specific mRNAs. However, their low abundance raises the possibility that other endogenous siRNAs may also participate. Together, our results demonstrate that maternal expression of Ago2 is essential for the earliest stages of mouse embryogenesis and are compatible with the notion that degradation of a proportion of maternal messages involves the RNAi-machinery.", "doi": "10.1091/mbc.e08-02-0219", "pmcid": "PMC2555945", "issn": "1059-1524", "publisher": "American Society for Cell Biology", "publication": "Molecular Biology of the Cell", "publication_date": "2008-10", "series_number": "10", "volume": "19", "issue": "10", "pages": "4383-4392" }, { "id": "authors:5vryy-pbr39", "collection": "authors", "collection_id": "5vryy-pbr39", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150055586", "type": "article", "title": "Bone morphogenetic protein 4 signaling regulates development of the anterior visceral endoderm in the mouse embryo", "author": [ { "family_name": "Soares", "given_name": "Miguel L.", "clpid": "Soares-M-L" }, { "family_name": "Torres-Padilla", "given_name": "Maria-Elena", "clpid": "Torres-Padilla-M-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The extraembryonic ectoderm (ExE) of the mouse conceptus is known to play a role in embryo patterning by signaling to the underlying epiblast and surrounding visceral endoderm. Bmp4 is one of the key ExE signaling molecules and has been recently implicated to participate in regulating development and migration of the anterior visceral endoderm (AVE). However, it remains unclear when exactly BMP4 signaling starts to regulate AVE positioning. To examine this, we have chosen to affect BMP4 function at two different time points, at embryonic day 5.25 (E5.25), thus before AVE migration, and E5.75, just after AVE migration. To this end, an RNAi technique was used, which consisted of the injection of Bmp4 dsRNA into the proamniotic cavity of the egg cylinder followed by its targeted electroporation into the ExE. This resulted in specific knockdown of Bmp4. It was found that Bmp4 RNAi at E5.25, but not at E5.75, led to an abnormal pattern of expression of the AVE marker Cerberus\u2010like. Thus, BMP4 signaling appears to affect the expression of Cer1 at a specific time window. This RNAi approach provides a convenient means to study spatial and temporal function of genes shortly after embryo implantation.", "doi": "10.1111/j.1440-169x.2008.01059.x", "pmcid": "PMC3342679", "issn": "0012-1592", "publisher": "Japanese Society of Developmental Biologists", "publication": "Development, Growth & Differentiation", "publication_date": "2008-09", "series_number": "7", "volume": "50", "issue": "7", "pages": "615-621" }, { "id": "authors:etr78-s6n52", "collection": "authors", "collection_id": "etr78-s6n52", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150055789", "type": "article", "title": "Pluripotency and differentiation in embryos and stem cells - Pavia, 17-18 January 2008", "author": [ { "family_name": "Adjaye", "given_name": "James A.", "clpid": "Adjaye-J-A" }, { "family_name": "Byskov", "given_name": "Anne G.", "clpid": "Byskov-A-G" }, { "family_name": "Cibelli", "given_name": "Jose B.", "clpid": "Cibelli-J-B" }, { "family_name": "De Maria", "given_name": "Ruggero", "clpid": "De-Maria-R" }, { "family_name": "Minger", "given_name": "Stephen", "clpid": "Minger-S" }, { "family_name": "Sampaolesi", "given_name": "Maurilio", "clpid": "Sampaolesi-M" }, { "family_name": "Testa", "given_name": "Giuseppe", "clpid": "Testa-G" }, { "family_name": "Verfaillie", "given_name": "Catherine", "clpid": "Verfaillie-C" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Sch\u00f6ler", "given_name": "Hans", "clpid": "Sch\u00f6ler-H-R" }, { "family_name": "Boiani", "given_name": "Michele", "clpid": "Boiani-M" }, { "family_name": "Crosetto", "given_name": "Nicola", "clpid": "Crosetto-N" }, { "family_name": "Redi", "given_name": "Carlo A.", "clpid": "Redi-C-A" } ], "abstract": "Each year many scientific meetings are held on stem cells to appraise the state of knowledge on their potency, differentiation and applications. So why did we hold another meeting? Because we thought one aspect was not adequately addressed in the others. When thinking of how our body is derived from a single fertilized egg, it is self-evident that the embryo is the 'mother' of all stem cells. This fact is probably overlooked because it is so remote (decades back in our lives!) and because embryonic stem cells do not exist as such in the embryo. However, this also tends to be ignored on purpose in many stem cell meetings because working on (human) embryos brings up substantial ethical concerns that bear on the scientific undertaking like nothing else. The origin of stem cells has become even more of a sensitive issue since the discovery in 2006 that embryonic stem (ES) cell-like cells can be generated in a petri dish straight from somatic cells by retrovirus-mediated transfer of selected genes. These new cells have been named 'induced pluripotent stem' (iPS) cells and have been obtained without any egg or embryo consumption (Takahashi and Yamanaka, 2006). This leads to the first topic of our meeting: natural and induced pluripotency...", "doi": "10.1387/ijdb.082695ja", "issn": "0214-6282", "publisher": "University of the Basque Country Press", "publication": "International Journal of Developmental Biology", "publication_date": "2008-07", "series_number": "7", "volume": "52", "issue": "7", "pages": "801-809" }, { "id": "authors:s34p4-ty362", "collection": "authors", "collection_id": "s34p4-ty362", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150055693", "type": "article", "title": "Formation of the embryonic-abembryonic axis of the mouse blastocyst: relationships between orientation of early cleavage divisions and pattern of symmetric/asymmetric divisions", "author": [ { "family_name": "Bischoff", "given_name": "Marcus", "clpid": "Bischoff-M" }, { "family_name": "Parfitt", "given_name": "David-Emlyn", "clpid": "Parfitt-D-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Setting aside pluripotent cells that give rise to the future body is a central cell fate decision in mammalian development. It requires that some blastomeres divide asymmetrically to direct cells to the inside of the embryo. Despite its importance, it is unknown whether the decision to divide symmetrically versus asymmetrically shows any spatial or temporal pattern, whether it is lineage-dependent or occurs at random, or whether it influences the orientation of the embryonic-abembryonic axis. To address these questions, we developed time-lapse microscopy to enable a complete 3D analysis of the origins, fates and divisions of all cells from the 2- to 32-cell blastocyst stage. This showed how in the majority of embryos, individual blastomeres give rise to distinct blastocyst regions. Tracking the division orientation of all cells revealed a spatial and temporal relationship between symmetric and asymmetric divisions and how this contributes to the generation of inside and outside cells and thus embryo patterning. We found that the blastocyst cavity, defining the abembryonic pole, forms where symmetric divisions predominate. Tracking cell ancestry indicated that the pattern of symmetric/asymmetric divisions of a blastomere can be influenced by its origin in relation to the animal-vegetal axis of the zygote. Thus, it appears that the orientation of the embryonic-abembryonic axis is anticipated by earlier cell division patterns. Together, our results suggest that two steps influence the allocation of cells to the blastocyst. The first step, involving orientation of 2- to 4-cell divisions along the animal-vegetal axis, can affect the second step, the establishment of inside and outside cell populations by asymmetric 8- to 32-cell divisions.", "doi": "10.1242/dev.014316", "pmcid": "PMC2655627", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "2008-03-01", "series_number": "5", "volume": "135", "issue": "5", "pages": "953-962" }, { "id": "authors:jmz4n-4z216", "collection": "authors", "collection_id": "jmz4n-4z216", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150055920", "type": "article", "title": "The anterior visceral endoderm of the mouse embryo is established from both preimplantation precursor cells and by de novo gene expression after implantation", "author": [ { "family_name": "Torres-Padilla", "given_name": "Maria-Elena", "clpid": "Torres-Padilla-M-E" }, { "family_name": "Richardson", "given_name": "Lucy", "clpid": "Richardson-L" }, { "family_name": "Kolasinska", "given_name": "Paulina", "clpid": "Kolasinska-P" }, { "family_name": "Meilhac", "given_name": "Sigol\u00e8ne M.", "clpid": "Meilhac-S-M" }, { "family_name": "Luetke-Eversloh", "given_name": "Merlin Verena", "clpid": "Luetke-Eversloh-M-V" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Initiation of the development of the anterior\u2013posterior axis in the mouse embryo has been thought to take place only when the anterior visceral endoderm (AVE) emerges and starts its asymmetric migration. However, expression of Lefty1, a marker of the AVE, was recently found to initiate before embryo implantation. This finding has raised two important questions: are the cells that show such early, preimplantation expression of this AVE marker the real precursors of the AVE and, if so, how does this contribute to the establishment of the AVE? Here, we address both of these questions. First, we show that the expression of another AVE marker, Cer1, also commences before implantation and its expression becomes consolidated in the subset of ICM cells that comprise the primitive endoderm. Second, to determine whether the cells showing this early Cer1 expression are true precursors of the AVE, we set up conditions to trace these cells in time-lapse studies from early periimplantation stages until the AVE emerges and becomes asymmetrically displaced. We found that Cer1-expressing cells are asymmetrically located after implantation and, as the embryo grows, they become dispersed into two or three clusters. The expression of Cer1 in the proximal domain is progressively diminished, whilst it is reinforced in the distal\u2013lateral domain. Our time-lapse studies demonstrate that this distal\u2013lateral domain is incorporated into the AVE together with cells in which Cer1 expression begins only after implantation. Thus, the AVE is formed from both part of an ancestral population of Cerl-expressing cells and cells that acquire Cer1 expression later. Finally, we demonstrate that when the AVE shifts asymmetrically to establish the anterior pole, this occurs towards the region where the earlier postimplantation expression of Cer1 was strongest. Together, these results suggest that the orientation of the anterior\u2013posterior axis is already anticipated before AVE migration.", "doi": "10.1016/j.ydbio.2007.06.020", "pmcid": "PMC3353121", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2007-09-01", "series_number": "1", "volume": "309", "issue": "1", "pages": "97-112" }, { "id": "authors:4aabe-2rg92", "collection": "authors", "collection_id": "4aabe-2rg92", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150056071", "type": "article", "title": "Regionalisation of the mouse visceral endoderm as the blastocyst transforms into the egg cylinder", "author": [ { "family_name": "Perea-Gomez", "given_name": "Aitana", "clpid": "Perea-Gomez-A" }, { "family_name": "Meilhac", "given_name": "Sigol\u00e8ne M.", "clpid": "Meilhac-S-M" }, { "family_name": "Piotrowska-Nitsche", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Gray", "given_name": "Dionne", "clpid": "Gray-D" }, { "family_name": "Collignon", "given_name": "J\u00e9r\u00f4me", "clpid": "Collignon-J" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Background: Reciprocal interactions between two extra-embryonic tissues, the extra-embryonic ectoderm and the visceral endoderm, and the pluripotent epiblast, are required for the establishment of anterior-posterior polarity in the mouse. After implantation, two visceral endoderm cell types can be distinguished, in the embryonic and extra-embryonic regions of the egg cylinder. In the embryonic region, the specification of the anterior visceral endoderm (AVE) is central to the process of anterior-posterior patterning. Despite recent advances in our understanding of the molecular interactions underlying the differentiation of the visceral endoderm, little is known about how cells colonise the three regions of the tissue. \n\nResults: As a first step, we performed morphological observations to understand how the extra-embryonic region of the egg cylinder forms from the blastocyst. Our analysis suggests a new model for the formation of this region involving cell rearrangements such as folding of the extra-embryonic ectoderm at the early egg cylinder stage. To trace visceral endoderm cells, we microinjected mRNAs encoding fluorescent proteins into single surface cells of the inner cell mass of the blastocyst and analysed the distribution of labelled cells at E5.0, E5.5 and E6.5. We found that at E5.0 the embryonic and extra-embryonic regions of the visceral endoderm do not correspond to distinct cellular compartments. Clusters of labelled cells may span the junction between the two regions even after the appearance of histological and molecular differences at E5.5. We show that in the embryonic region cell dispersion increases after the migration of the AVE. At this time, visceral endoderm cell clusters tend to become oriented parallel to the junction between the embryonic and extra-embryonic regions. Finally we investigated the origin of the AVE and demonstrated that this anterior signalling centre arises from more than a single precursor between E3.5 and E5.5. \n\nConclusion: We propose a new model for the formation of the extra-embryonic region of the egg cylinder involving a folding of the extra-embryonic ectoderm. Our analyses of the pattern of labelled visceral endoderm cells indicate that distinct cell behaviour in the embryonic and extra-embryonic regions is most apparent upon AVE migration. We also demonstrate the polyclonal origin of the AVE. Taken together, these studies lead to further insights into the formation of the extra-embryonic tissues as they first develop after implantation.", "doi": "10.1186/1471-213x-7-96", "pmcid": "PMC1978209", "issn": "1471-213X", "publisher": "BioMed Central", "publication": "BMC Developmental Biology", "publication_date": "2007-08-16", "volume": "7", "pages": "Art. No. 96" }, { "id": "authors:4xd95-pf107", "collection": "authors", "collection_id": "4xd95-pf107", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150056179", "type": "article", "title": "Novel gene expression patterns along the proximo-distal axis of the mouse embryo before gastrulation", "author": [ { "family_name": "Frankenberg", "given_name": "Stephen", "clpid": "Frankenberg-S" }, { "family_name": "Smith", "given_name": "Lee", "clpid": "Smith-Lee" }, { "family_name": "Greenfield", "given_name": "Andy", "clpid": "Greenfield-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Background: To date, the earliest stage at which the orientation of the anterior-posterior axis in the mouse embryo is distinguishable by asymmetric gene expression is shortly after E5.5. At E5.5, prospective anterior markers are expressed at the distal tip of the embryo, whereas prospective posterior markers are expressed more proximally, close to the boundary with the extraembryonic region. \n\nResults: To contribute to elucidating the mechanisms underlying the events involved in early patterning of the mouse embryo, we have carried out a microarray screen to identify novel genes that are differentially expressed between the distal and proximal parts of the E5.5 embryo. Secondary screening of resulting candidates by in situ hybridisation at E5.5 and E6.5 revealed novel expression patterns for known and previously uncharacterised genes, including Peg10, Ctsz1, Cubilin, Jarid1b, Ndrg1, Sfmbt2, Gjb5, Talia and Plet1. The previously undescribed gene Talia and recently identified Plet1 are expressed specifically in the distal-most part of the extraembryonic ectoderm, adjacent to the epiblast, and are therefore potential candidates for regulating early patterning events. Talia and the previously described gene XE7 define a gene family highly conserved among metazoans and with a predicted protein structure suggestive of a post-transcriptional regulative function, whilst Plet1 appears to be mammal-specific and of unknown function. \n\nConclusion: Our approach has allowed us to compare expression between dissected parts of the egg cylinder and has identified multiple genes with novel expression patterns at this developmental stage. These genes are potential candidates for regulating tissue interactions following implantation.", "doi": "10.1186/1471-213x-7-8", "pmcid": "PMC1821012", "issn": "1471-213X", "publisher": "BioMed Central", "publication": "BMC Developmental Biology", "publication_date": "2007-02-15", "volume": "7", "pages": "Art. No. 8" }, { "id": "authors:wgm60-rcm73", "collection": "authors", "collection_id": "wgm60-rcm73", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190418-150056272", "type": "article", "title": "Dishevelled proteins regulate cell adhesion in mouse blastocyst and serve to monitor changes in Wnt signaling", "author": [ { "family_name": "Na", "given_name": "Jie", "clpid": "Na-Jie" }, { "family_name": "Lykke-Andersen", "given_name": "Karin", "clpid": "Lykke-Andersen-K" }, { "family_name": "Torres Padilla", "given_name": "Maria Elena", "clpid": "Torres-Padilla-M-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Wnt signaling is essential for the regulation of cell polarity and cell fate in the early embryogenesis of many animal species. Multiple Wnt genes and its pathway members are expressed in the mouse early embryo, raising the question whether they play any roles in preimplantation development. Dishevelled is an important transducer of divergent Wnt pathways. Here we show that three of the mouse Dishevelled proteins are not only expressed in oocytes and during preimplantation development, but also display distinct spatio-temporal localization. Interestingly, as embryos reach blastocyst stage, Dishevelled 2 becomes increasingly associated with cell membrane in trophectoderm cells, while at E4.5, Dishevelled 3 is highly enriched in the cytoplasm of ICM cells. These changes are coincident with an increase in the active form of \u03b2-catenin, p120catenin transcription and decrease of Kaiso expression, indicating an upregulation of Wnt signaling activity before implantation. When Dishevelled-GFP fusion proteins are overexpressed in single blastomeres of the 4-cell stage embryo, the progeny of this cell show reduction in cell adhesiveness and a rounded shape at the blastocyst stage. This suggests that perturbing Dvl function interferes with cell\u2013cell adhesion through the non-canonical Wnt pathway in blastocysts.", "doi": "10.1016/j.ydbio.2006.08.036", "pmcid": "PMC3353122", "issn": "0012-1606", "publisher": "Elsevier", "publication": "Developmental Biology", "publication_date": "2007-02-01", "series_number": "1", "volume": "302", "issue": "1", "pages": "40-49" }, { "id": "authors:5h17j-kw650", "collection": "authors", "collection_id": "5h17j-kw650", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105736503", "type": "article", "title": "Histone arginine methylation regulates pluripotency in the early mouse embryo", "author": [ { "family_name": "Torres-Padilla", "given_name": "Maria-Elena", "clpid": "Torres-Padilla-M-E" }, { "family_name": "Parfitt", "given_name": "David-Emlyn", "clpid": "Parfitt-D-E" }, { "family_name": "Kouzarides", "given_name": "Tony", "clpid": "Kouzarides-T" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "It has been generally accepted that the mammalian embryo starts its development with all cells identical, and only when inside and outside cells form do differences between cells first emerge. However, recent findings show that cells in the mouse embryo can differ in their developmental fate and potency as early as the four-cell stage1,2,3,4. These differences depend on the orientation and order of the cleavage divisions that generated them2,5. Because epigenetic marks are suggested to be involved in sustaining pluripotency6,7, we considered that such developmental properties might be achieved through epigenetic mechanisms. Here we show that modification of histone H3, through the methylation of specific arginine residues, is correlated with cell fate and potency. Levels of H3 methylation at specific arginine residues are maximal in four-cell blastomeres that will contribute to the inner cell mass (ICM) and polar trophectoderm and undertake full development when combined together in chimaeras. Arginine methylation of H3 is minimal in cells whose progeny contributes more to the mural trophectoderm and that show compromised development when combined in chimaeras. This suggests that higher levels of H3 arginine methylation predispose blastomeres to contribute to the pluripotent cells of the ICM. We confirm this prediction by overexpressing the H3-specific arginine methyltransferase CARM1 in individual blastomeres and show that this directs their progeny to the ICM and results in a dramatic upregulation of Nanog and Sox2. Thus, our results identify specific histone modifications as the earliest known epigenetic marker contributing to development of ICM and show that manipulation of epigenetic information influences cell fate determination.", "doi": "10.1038/nature05458", "pmcid": "PMC3353120", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2007-01-11", "series_number": "7124", "volume": "445", "issue": "7124", "pages": "214-218" }, { "id": "authors:5enbg-7p012", "collection": "authors", "collection_id": "5enbg-7p012", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105736662", "type": "article", "title": "The first cell-fate decisions in the mouse embryo: destiny is a matter of both chance and choice", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Development of the early mouse embryo has always been classified as regulative, meaning that when parts or blastomeres of the embryo are isolated they change their developmental fate and can even reconstruct the whole. However, regulative development does not mean that, in situ, these parts or blastomeres are equivalent; it does not mean that the early mammalian embryo is a ball of identical cells without any bias. Regulative development simply means that whatever bias the regions of the embryo might have they still remain flexible and can respond to experimental interference by changes of fate. This realization \u2014 that regulative development and patterning can co-exist \u2014 has led to a renaissance of interest in the first days of development of the mouse embryo, and several laboratories have provided evidence for some early bias. Now the challenge is to gain some understanding of the molecular basis of this bias.", "doi": "10.1016/j.gde.2006.06.011", "issn": "0959-437X", "publisher": "Current Biology Ltd", "publication": "Current Opinion in Genetics and Development", "publication_date": "2006-08", "series_number": "4", "volume": "16", "issue": "4", "pages": "406-412" }, { "id": "authors:7nq9s-52902", "collection": "authors", "collection_id": "7nq9s-52902", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105736755", "type": "article", "title": "Role of TIF1\u03b1 as a modulator of embryonic transcription in the mouse zygote", "author": [ { "family_name": "Torres-Padilla", "given_name": "Maria Elena", "clpid": "Torres-Padilla-M-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The first events of the development of any embryo are under maternal control until the zygotic genome becomes activated. In the mouse embryo, the major wave of transcription activation occurs at the 2-cell stage, but transcription starts already at the zygote (1-cell) stage. Very little is known about the molecules involved in this process. We show that the transcription intermediary factor 1 \u03b1 (TIF1\u03b1) is involved in modulating gene expression during the first wave of transcription activation. At the onset of genome activation, TIF1\u03b1 translocates from the cytoplasm into the pronuclei to sites of active transcription. These sites are enriched with the chromatin remodelers BRG-1 and SNF2H. When we ablate TIF1\u03b1 through either RNA interference (RNAi) or microinjection of specific antibodies into zygotes, most of the embryos arrest their development at the 2\u20134-cell stage transition. The ablation of TIF1\u03b1 leads to mislocalization of RNA polymerase II and the chromatin remodelers SNF2H and BRG-1. Using a chromatin immunoprecipitation cloning approach, we identify genes that are regulated by TIF1\u03b1 in the zygote and find that transcription of these genes is misregulated upon TIF1\u03b1 ablation. We further show that the expression of some of these genes is dependent on SNF2H and that RNAi for SNF2H compromises development, suggesting that TIF1\u03b1 mediates activation of gene expression in the zygote via SNF2H. These studies indicate that TIF1\u03b1 is a factor that modulates the expression of a set of genes during the first wave of genome activation in the mouse embryo.", "doi": "10.1083/jcb.200603146", "pmcid": "PMC2064229", "issn": "0021-9525", "publisher": "Rockefeller University Press", "publication": "Journal of Cell Biology", "publication_date": "2006-07-31", "series_number": "3", "volume": "174", "issue": "3", "pages": "329-338" }, { "id": "authors:xc536-knb84", "collection": "authors", "collection_id": "xc536-knb84", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105736846", "type": "article", "title": "Asymmetric Positioning and Organization of the Meiotic Spindle of Mouse Oocytes Requires CDC42 Function", "author": [ { "family_name": "Na", "given_name": "Jie", "clpid": "Na-Jie" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The mature mammalian oocyte is highly polarized because asymmetrical spindle migration to the oocyte cortex ensures extrusion of small polar bodies in the two meiotic divisions, essential for generation of the large egg. Actin filaments, myosin motors, and formin-2, but not microtubules, are required for spindle migration 1, 2, 3. Here, we show that Cdc42, a key regulator of cytoskeleton and cell polarity in other systems 4, 5, 6, 7, 8, 9, 10, is essential for meiotic maturation and oocyte asymmetry. Disrupting CDC42 function by ectopic expression of its GTPase-defective mutants causes both halves of the first meiotic spindle to extend symmetrically toward opposing cortical regions and prevents an asymmetrical division. The elongated spindle has numerous astral-like microtubules, and aPKC\u03b6, normally associated with the spindle poles, is distributed along its length. Dynactin is displaced from kinetochores, consistently homologous chromosomes do not segregate, and polar body extrusion is prevented. Perturbing the function of aPKC\u03b6 also causes elongation of the meiotic spindle but still permits spindle migration and polar body extrusion. Thus, at least two pathways appear to be downstream of CDC42: one affecting the actin cytoskeleton and required for migration of the meiotic spindle, and a second affecting the spindle microtubules in which aPKC\u03b6 plays a role.", "doi": "10.1016/j.cub.2006.05.023", "issn": "0960-9822", "publisher": "Cell Press", "publication": "Current Biology", "publication_date": "2006-06-20", "series_number": "12", "volume": "16", "issue": "12", "pages": "1249-1254" }, { "id": "authors:hsbyk-9t796", "collection": "authors", "collection_id": "hsbyk-9t796", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105736940", "type": "article", "title": "Regionalised signalling within the extraembryonic ectoderm regulates anterior visceral endoderm positioning in the mouse embryo", "author": [ { "family_name": "Richardson", "given_name": "Lucy", "clpid": "Richardson-L" }, { "family_name": "Torres-Padilla", "given_name": "Maria-Elena", "clpid": "Torres-Padilla-M-E" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The development of the anterior\u2013posterior (AP) axis in the mammalian embryo is controlled by interactions between embryonic and extraembryonic tissues. It is well established that one of these extraembryonic tissues, the anterior visceral endoderm (AVE), can repress posterior cell fate and that signalling from the other, the extraembryonic ectoderm (ExE), is required for posterior patterning. Here, we show that signals from the prospective posterior ExE repress AVE gene expression and affect the distribution of the AVE cells. Surgical ablation of the prospective posterior, but not the anterior, extraembryonic region at 5.5 days of development (E5.5) perturbs the characteristic distal-to-anterior distribution of AVE cells and leads to a dramatic expansion of the AVE domain. Time-lapse imaging studies show that this increase is due to the ectopic expression of an AVE marker, which results in a symmetrical positioning of the AVE. Surgical ablation of this same ExE region after the distal-to-anterior migration has already commenced, at E5.75, does not affect the localisation of the AVE, indicating that this effect takes place within a short time window. Conversely, transplanting the prospective posterior, but not the anterior, extraembryonic region onto isolated E5.5 embryonic explants drastically reduces the AVE domain. Further, transplantation experiments demonstrate that the signalling regulating AVE gene expression originates from the posterior ExE, rather than its surrounding VE. Together, our results show that signals emanating from the future posterior ExE within a temporal window both restrict the AVE domain and promote its specific positioning. This indicates for the first time that the ExE is already regionalised a day before the onset of gastrulation in order to correctly set the orientation of the AP axis of the mouse embryo. We propose a reciprocal function of the posterior ExE and the AVE in establishing a balance between the antagonistic activities of these two tissues, essential for AP patterning.", "doi": "10.1016/j.mod.2006.01.004", "issn": "0925-4773", "publisher": "Elsevier", "publication": "Mechanisms of Development", "publication_date": "2006-04", "series_number": "4", "volume": "123", "issue": "4", "pages": "288-296" }, { "id": "authors:mpq56-a6t73", "collection": "authors", "collection_id": "mpq56-a6t73", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105737033", "type": "article", "title": "Dynamic distribution of the replacement histone variant H3.3 in the mouse oocyte and preimplantation embryos", "author": [ { "family_name": "Torres-Padilla", "given_name": "Maria-Elena", "clpid": "Torres-Padilla-M-E" }, { "family_name": "Bannister", "given_name": "Andrew J.", "clpid": "Bannister-A-J" }, { "family_name": "Hurd", "given_name": "Paul J.", "clpid": "Hurd-P-J" }, { "family_name": "Kouzarides", "given_name": "Tony", "clpid": "Kouzarides-T" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Upon fertilization, the gametes undergo a drastic reprogramming that includes changes in DNA methylation and histone modifications. Currently, it is not known whether replacement of the major histones by histone variants is also involved in these processes. Here we have examined the expression and localization of the histone variant H3.3 in early mouse embryogenesis. We show that H3.3 is present in the oocyte as a maternal factor. It is then incorporated preferentially into the male pronucleus before genome activation, pointing towards an asymmetry in histone composition between the two pronuclei. This is in line with the male pronucleus bearing transcriptional activation first. The same distribution was observed when we followed the localisation of a tagged version of H3.3. We detected H3.3 in the nuclei of mouse embryos in all of the stages analysed, from the zygote to the blastocyst stage, suggesting that the epigenetic mechanisms in the early embryo not only involve changes in histone modifications but may also include histone replacement.", "doi": "10.1387/ijdb.052073mt", "issn": "0214-6282", "publisher": "University of the Basque Country Press", "publication": "International Journal of Developmental Biology", "publication_date": "2006-01", "series_number": "5", "volume": "50", "issue": "5", "pages": "455-461" }, { "id": "authors:7w824-sph11", "collection": "authors", "collection_id": "7w824-sph11", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105737131", "type": "article", "title": "Functional studies of signaling pathways in peri-implantation development of the mouse embryo by RNAi", "author": [ { "family_name": "Soares", "given_name": "Miguel L.", "clpid": "Soares-M-L" }, { "family_name": "Haraguchi", "given_name": "Seiki", "clpid": "Haraguchi-Seiki" }, { "family_name": "Torres-Padilla", "given_name": "Maria-Elena", "clpid": "Torres-Padilla-M-E" }, { "family_name": "Kalmar", "given_name": "Tibor", "clpid": "Kalmar-T" }, { "family_name": "Carpenter", "given_name": "Lee", "clpid": "Carpenter-L" }, { "family_name": "Bell", "given_name": "Graham", "clpid": "Bell-G" }, { "family_name": "Morrison", "given_name": "Alastair", "clpid": "Morrison-A" }, { "family_name": "Ring", "given_name": "Christopher J. A.", "clpid": "Ring-C-J-A" }, { "family_name": "Clarke", "given_name": "Neil J.", "clpid": "Clarke-N-J" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Background: Studies of gene function in the mouse have relied mainly on gene targeting via homologous recombination. However, this approach is difficult to apply in specific windows of time, and to simultaneously knock-down multiple genes. Here we report an efficient method for dsRNA-mediated gene silencing in late cleavage-stage mouse embryos that permits examination of phenotypes at post-implantation stages. \n\nResults: We show that introduction of Bmp4 dsRNA into intact blastocysts by electroporation recapitulates the genetic Bmp4 null phenotype at gastrulation. It also reveals a novel role for Bmp4 in the regulation the anterior visceral endoderm specific gene expression and its positioning. We also show that RNAi can be used to simultaneously target several genes. When applied to the three murine isoforms of Dishevelled, it leads to earlier defects than previously observed in double knock-outs. These include severe delays in post-implantation development and defects in the anterior midline and neural folds at headfold stages. \n\nConclusion: Our results indicate that the BMP4 signalling pathway contributes to the development of the anterior visceral endoderm, and reveal an early functional redundancy between the products of the murine Dishevelled genes. The proposed approach constitutes a powerful tool to screen the functions of genes that govern the development of the mouse embryo.", "doi": "10.1186/1471-213x-5-28", "pmcid": "PMC1363358", "issn": "1471-213X", "publisher": "BioMed Central", "publication": "BMC Developmental Biology", "publication_date": "2005-12-28", "volume": "5", "pages": "Art. No. 28" }, { "id": "authors:qfnpe-q9g70", "collection": "authors", "collection_id": "qfnpe-q9g70", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105737517", "type": "article", "title": "Cleavage pattern and emerging asymmetry of the mouse embryo", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Early mammalian development is regulative \u2014 it is flexible and responsive to experimental intervention. This flexibility could be explained if embryogenesis were originally completely unbiased and disordered; order and determination of cells only arising later. Alternatively, regulative behaviour could be consistent with the embryo having some order or bias from the very beginning, with inflexibility and cell determination increasing steadily over time. Recent evidence supports the second view and indicates that the sequence and the orientations of cell divisions help to build the first asymmetries.", "doi": "10.1038/nrm1782", "issn": "1471-0072", "publisher": "Nature Publishing Group", "publication": "Nature Reviews. Molecular Cell Biology", "publication_date": "2005-12", "series_number": "12", "volume": "6", "issue": "12", "pages": "919-928" }, { "id": "authors:z1rm9-ysx56", "collection": "authors", "collection_id": "z1rm9-ysx56", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105737609", "type": "article", "title": "PAR-1 and the microtubule-associated proteins CLASP2 and dynactin-p50 have specific localisation on mouse meiotic and first mitotic spindles", "author": [ { "family_name": "Moore", "given_name": "Catherine A.", "clpid": "Moore-C-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The site of second meiotic division, marked by the second polar body, is an important reference point in the early mouse embryo. To study its formation, we look at the highly asymmetric meiotic divisions. For extrusion of the small polar bodies during meiosis, the spindles must be located cortically. The positioning of meiotic spindles is known to involve the actin cytoskeleton, but whether microtubules are also involved is not clear. In this study we investigated the patterns of localisation of microtubule regulatory proteins in mouse oocytes. PAR-1 is a member of the PAR (partitioning-defective) family with known roles in regulation of microtubule stability and spindle positioning in other model systems. Here we show its specific localisation on mouse meiotic and first mitotic spindles. In addition, the microtubule-associated proteins CLASP2 (a CLIP associating protein) and dynactin-p50 are found on kinetochores and a subset of microtubule-organising centres. Thus we show specific localisation of microtubule regulatory proteins in mouse oocytes, which could indicate roles in meiotic spindle organisation.", "doi": "10.1530/rep.1.00651", "issn": "1470-1626", "publisher": "Bioscientifica", "publication": "Reproduction", "publication_date": "2005-09", "series_number": "3", "volume": "130", "issue": "3", "pages": "311-320" }, { "id": "authors:05w6a-ge923", "collection": "authors", "collection_id": "05w6a-ge923", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105737708", "type": "article", "title": "Spatial arrangement of individual 4-cell stage blastomeres and the order in which they are generated correlate with blastocyst pattern in the mouse embryo", "author": [ { "family_name": "Piotrowska-Nitsche", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "In the unperturbed development of the mouse embryo one of the 2-cell blastomeres tends to contribute its progeny predominantly to the embryonic and the other to the abembryonic part of the blastocyst. However, a significant minority of embryos (20\u201330%) do not show this correlation. In this study, we have used non-invasive lineage tracing to determine whether development of blastocyst pattern shows any correlation with the orientation and order of the second cleavage divisions that result in specific positioning of blastomeres at the 4-cell stage. Although the orientation and order of the second cleavages are not predetermined, in the great majority (80%) of embryos the spatial arrangement of 4-cell blastomeres is consistent with one of the second cleavages occurring meridionally and the other equatorially or obliquely with respect to the polar body. In such cleaving embryos, one of the 2-cell stage blastomeres tends to contribute to embryonic while the other contributes predominantly to abembryonic part of the blastocyst. Thus, in these embryos the outcome of the first cleavage tends to correlate with the orientation of the blastocyst embryonic\u2013abembryonic axis. However, the order of blastomere divisions predicts a specific polarity for this axis only when the earlier 2-cell blastomere to divide does so meridionally. In contrast to the above two groups, in those embryos in which both second cleavage divisions occur in a similar orientation, either meridionally or equatorially, we do not observe any tendency for the 2-cell blastomeres to contribute to specific blastocyst parts. We find that all these groups of embryos develop to term with similar success, with the exception of those in which both second cleavage divisions occur equatorially whose development can be compromised. We conclude that the orientations and order of the second cleavages are not predetermined; they correlate with the development of blastocyst patterning; and that the majority, but not all, of these cleavage patterns allow equally successful development.", "doi": "10.1016/j.mod.2004.11.014", "issn": "0925-4773", "publisher": "Elsevier", "publication": "Mechanisms of Development", "publication_date": "2005-04", "series_number": "4", "volume": "122", "issue": "4", "pages": "487-500" }, { "id": "authors:bbw29-yh191", "collection": "authors", "collection_id": "bbw29-yh191", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105737803", "type": "article", "title": "The first cleavage of the mouse zygote predicts the blastocyst axis", "author": [ { "family_name": "Plusa", "given_name": "Berenika", "clpid": "Plusa-B" }, { "family_name": "Hadjantonakis", "given_name": "Anna-Katerina", "clpid": "Hadjantonakis-A-K" }, { "family_name": "Gray", "given_name": "Dionne", "clpid": "Gray-D" }, { "family_name": "Piotrowska-Nitsche", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Jedrusik", "given_name": "Agnieszka", "clpid": "Jedrusik-A" }, { "family_name": "Papaioannou", "given_name": "Virginia E.", "clpid": "Papaioannou-V-E" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "One of the unanswered questions in mammalian development is how the embryonic\u2013abembryonic axis of the blastocyst is first established. It is possible that the first cleavage division contributes to this process, because in most mouse embryos the progeny of one two-cell blastomere primarily populate the embryonic part of the blastocyst and the progeny of its sister populate the abembryonic part1,2,3,4. However, it is not known whether the embryonic\u2013abembryonic axis is set up by the first cleavage itself, by polarity in the oocyte that then sets the first cleavage plane with respect to the animal pole, or indeed whether it can be divorced entirely from the first cleavage and established in relation to the animal pole. Here we test the importance of the orientation of the first cleavage by imposing an elongated shape on the zygote so that the division no longer passes close to the animal pole, marked by the second polar body. Non-invasive lineage tracing shows that even when the first cleavage occurs along the short axis imposed by this experimental treatment, the progeny of the resulting two-cell blastomeres tend to populate the respective embryonic and abembryonic parts of the blastocyst. Thus, the first cleavage contributes to breaking the symmetry of the embryo, generating blastomeres with different developmental characteristics.", "doi": "10.1038/nature03388", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2005-03-17", "series_number": "7031", "volume": "434", "issue": "7031", "pages": "391-395" }, { "id": "authors:pw1xe-etz31", "collection": "authors", "collection_id": "pw1xe-etz31", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105738009", "type": "article", "title": "Downregulation of Par3 and aPKC function directs cells towards the ICM in the preimplantation mouse embryo", "author": [ { "family_name": "Plusa", "given_name": "Berenika", "clpid": "Plusa-B" }, { "family_name": "Frankenberg", "given_name": "Stephen", "clpid": "Frankenberg-S" }, { "family_name": "Chalmers", "given_name": "Andrew", "clpid": "Chalmers-A" }, { "family_name": "Hadjantonakis", "given_name": "Anna-Katerina", "clpid": "Hadjantonakis-A-K" }, { "family_name": "Moore", "given_name": "Catherine A.", "clpid": "Moore-C-A" }, { "family_name": "Papalopulu", "given_name": "Nancy", "clpid": "Papalopulu-N" }, { "family_name": "Papaioannou", "given_name": "Virginia E.", "clpid": "Papaioannou-V-E" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Generation of inside cells that develop into inner cell mass (ICM) and outside cells that develop into trophectoderm is central to the development of the early mouse embryo. Critical to this decision is the development of cell polarity and the associated asymmetric (differentiative) divisions of the 8-cell-stage blastomeres. The underlying molecular mechanisms for these events are not understood. As the Par3/aPKC complex has a role in establishing cellular polarity and division orientation in other systems, we explored its potential function in the developing mouse embryo. We show that both Par3 and aPKC adopt a polarized localization from the 8-cell stage onwards and that manipulating their function re-directs cell positioning and consequently influences cell fate. Injection of dsRNA against Par3 or mRNA for a dominant negative form of aPKC into a random blastomere at the 4-cell stage directs progeny of the injected cell into the inside part of the embryo. This appears to result from both an increased frequency by which such cells undertake differentiative divisions and their decreased probability of retaining outside positions. Thus, the natural spatial allocation of blastomere progeny can be over-ridden by downregulation of Par3 or aPKC, leading to a deceased tendency for them to remain outside and so develop into trophectoderm. In addition, this experimental approach illustrates a powerful means of manipulating gene expression in a specific clonal population of cells in the preimplantation embryo.", "doi": "10.1242/jcs.01666", "issn": "0021-9533", "publisher": "Company of Biologists", "publication": "Journal of Cell Science", "publication_date": "2005-02-01", "series_number": "3", "volume": "118", "issue": "3", "pages": "505-515" }, { "id": "authors:gyh59-mwf38", "collection": "authors", "collection_id": "gyh59-mwf38", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105737904", "type": "article", "title": "Four-cell stage mouse blastomeres have different developmental properties", "author": [ { "family_name": "Piotrowska-Nitsche", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Perea-Gomez", "given_name": "Aitana", "clpid": "Perea-Gomez-A" }, { "family_name": "Haraguchi", "given_name": "Seiki", "clpid": "Haraguchi-Seiki" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Blastomeres of the early mouse embryo are thought to be equivalent in their developmental properties at least until the eight-cell stage. However, the experiments that have led to this conclusion could not have taken into account either the spatial origin of individual blastomeres or the spatial allocation and fate of their progeny. We have therefore readdressed this issue having defined cell lineages in mouse embryos undergoing different patterns of cleavage in their second division cycle. This has enabled us to identify a major group of embryos in which we can predict not only the spatial origin of each given four-cell blastomeres, but also which region of the blastocyst is most likely to be occupied by its progeny. We show that a pattern of second cleavage divisions in which a meridional division is followed by one that is equatorial or oblique allows us to identify blastomeres that differ in their fate and in their developmental properties both from each other and from their cousins. We find that one of these four-cell stage blastomeres that inherits some vegetal membrane marked in the previous cleavage cycle tends to contribute to mural trophectoderm. The progeny of its sister tend to donate cells to part of the ICM lining the blastocyst cavity and its associated trophectoderm. Chimaeras made entirely of these equatorially or obliquely derived blastomeres show developmental abnormalities in both late preimplantation and early postimplantation development. By contrast, chimaeras made from four-cell stage blastomeres from early meridional divisions develop normally. The developmental defects of chimaeras made from the most vegetal blastomeres that result from later second cleavages are the most severe and following transplantation into foster mothers they fail to develop to term. However, when such individual four-cell blastomeres are surrounded by blastomeres from random positions, they are able to contribute to all embryonic lineages. In conclusion, this study shows that while all four-cell blastomeres can have full developmental potential, they differ in their individual developmental properties according to their origin in the embryo from as early as the four-cell stage.", "doi": "10.1242/dev.01602", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "2005-02", "series_number": "3", "volume": "132", "issue": "3", "pages": "479-490" }, { "id": "authors:256zs-52d38", "collection": "authors", "collection_id": "256zs-52d38", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105738149", "type": "article", "title": "Directing pluripotent cell differentiation using \"diced RNA\" in transient transfection", "author": [ { "family_name": "Carpenter", "given_name": "Lee", "clpid": "Carpenter-L" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Embryonic stem (ES) and embryonic carcinoma (EC) cells are pluripotent and have the capacity to differentiate into many cell types. The ability to direct their differentiation should have considerable practical applications. Here, we first report the use of diced short interfering RNAi against Oct4 in a transient approach, to direct differentiation of ES towards the trophectoderm lineage. We then apply this approach to downregulate Smad4 in mouse P19 EC cells. We have found that this leads to an increase in the levels of Pax6 (a neuroectoderm marker), reduction in the levels of Brachyury (a mesoderm marker), and a 3\u2010fold increase in the number of \u03b2III tubulin\u2010positive colonies when these cells were allowed to differentiate. This indicates a redirection of cell fate towards the neuroectoderm lineage. Thus, transient RNAi could provide a valuable tool to direct pluripotent cells along specific pathways of differentiation while circumventing permanent genetic changes.", "doi": "10.1002/gene.20078", "issn": "1526-954X", "publisher": "Wiley", "publication": "Genesis", "publication_date": "2004-11", "series_number": "3", "volume": "40", "issue": "3", "pages": "157-163" }, { "id": "authors:cdf0f-f5x20", "collection": "authors", "collection_id": "cdf0f-f5x20", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105738244", "type": "article", "title": "First cell fate decisions and spatial patterning in the early mouse embryo", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "A growing body of evidence indicates that although the early mouse embryo retains flexibility in responding to perturbations, its patterning is initiated at the earliest developmental stages. There are a few spatial cues that are able to influence the pattern of cleavage divisions: one of these lies in the vicinity of the previous meiotic division, the second is associated with the sperm entry and, related to this, the third is the cell shape. Furthermore, the first cleavage separates the zygote into two cells that tend to follow distinguishable fates: one contributes mainly to the embryonic part of the blastocyst, and the other to the abembryonic. The cumulative effect of the early asymmetries generated through cleavage might lead to asymmetric interactions between the first lineages of cells. This could influence development of patterning after implantation. These early polarity cues serve to bias patterning and not as definitive determinants.", "doi": "10.1016/j.semcdb.2004.04.004", "issn": "1084-9521", "publisher": "Elsevier", "publication": "Seminars in Cell and Developmental Biology", "publication_date": "2004-10", "series_number": "5", "volume": "15", "issue": "5", "pages": "563-572" }, { "id": "authors:s9v51-wz018", "collection": "authors", "collection_id": "s9v51-wz018", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105738331", "type": "article", "title": "First Cleavage of the Mouse Embryo Responds to Change in Egg Shape at Fertilization", "author": [ { "family_name": "Gray", "given_name": "Dionne", "clpid": "Gray-D" }, { "family_name": "Plusa", "given_name": "Berenika", "clpid": "Plusa-B" }, { "family_name": "Piotrowska", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Na", "given_name": "Jie", "clpid": "Na-Jie" }, { "family_name": "Tom", "given_name": "Brian", "clpid": "Tom-B-D" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Although mouse development is regulative, the cleavage pattern of the embryo is not random 1, 2. The first cleavage tends to relate to the site of the previous meiosis 3, 4. Sperm entry might provide a second cue, but evidence for 5, 6 and against 7, 8 this is indirect and has been debated 9, 10. To resolve whether sperm entry position relates to the first cleavage, we have followed development from fertilization by time-lapse imaging. This directly showed cytokinesis passes close to the site of the previous meiosis and to both the sperm entry site and trajectory of the male pronucleus in a significant majority of eggs. We detected asymmetric distribution of Par6 protein in relation to the site of meiosis, but not sperm entry. Unexpectedly, we found the egg becomes flattened upon fertilization in an actin-mediated process. The sperm entry position tends to lie at one end of the short axis along which cleavage will pass. When we manipulated eggs to change their shape, this repositioned the cleavage plane such that eggs divided along their experimentally imposed short axis. Such manipulated eggs were able to develop to term, emphasizing the regulative nature of their development.", "doi": "10.1016/j.cub.2004.02.031", "issn": "0960-9822", "publisher": "Cell Press", "publication": "Current Biology", "publication_date": "2004-03-09", "series_number": "5", "volume": "14", "issue": "5", "pages": "397-405" }, { "id": "authors:adfsm-22897", "collection": "authors", "collection_id": "adfsm-22897", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105738498", "type": "article", "title": "The Anterior-Posterior Axis Emerges Respecting the Morphology of the Mouse Embryo that Changes and Aligns with the Uterus before Gastrulation", "author": [ { "family_name": "Mesnard", "given_name": "Daniel", "clpid": "Mesnard-D" }, { "family_name": "Filipe", "given_name": "Mario", "clpid": "Filipe-M" }, { "family_name": "Belo", "given_name": "Jos\u00e9 A.", "clpid": "Belo-J-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Background: When the anterior-posterior axis of the mouse embryo becomes explicit at gastrulation, it is almost perpendicular to the long uterine axis. This led to the belief that the uterus could play a key role in positioning this future body axis. \n\nResults: Here, we demonstrate that when the anterior-posterior axis first emerges it does not respect the axes of the uterus but, rather, the morphology of the embryo. Unexpectedly, the emerging anterior-posterior axis is initially aligned not with the long, but the short axis of the embryo. Then whether the embryo develops in vitro or in utero, the anterior-posterior axis becomes aligned with the long axis of embryo just prior to gastrulation. Of three mechanisms that could account for this apparent shift in anterior-posterior axis orientation\u2013cell migration, spatial change of gene expression, or change in embryo shape\u2013lineage tracing studies favor a shape change accompanied by restriction of the expression domain of anterior markers. This property of the embryo must be modulated by interactions with the uterus as ultimately the anterior-posterior and long axes of the embryo align with the left-right uterine axis. \n\nConclusions: The emerging anterior-posterior axis relates to embryo morphology rather than that of the uterus. The apparent shift in its orientation to align with the long embryonic axis and with the uterus is associated with a change in embryo shape and a refinement of anterior gene expression pattern. This suggests an interdependence between anterior-posterior gene expression, the shape of the embryo, and the uterus.", "doi": "10.1016/j.cub.2004.01.026", "issn": "0960-9822", "publisher": "Cell Press", "publication": "Current Biology", "publication_date": "2004-02-03", "series_number": "3", "volume": "14", "issue": "3", "pages": "184-196" }, { "id": "authors:k9fay-rtg73", "collection": "authors", "collection_id": "k9fay-rtg73", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105738586", "type": "article", "title": "A Genome-Wide Study of Gene Activity Reveals Developmental Signaling Pathways in the Preimplantation Mouse Embryo", "author": [ { "family_name": "Wang", "given_name": "Q. Tian", "clpid": "Wang-Q-Tian" }, { "family_name": "Piotrowska", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Ciemerych", "given_name": "Maria Anna", "clpid": "Ciemerych-M-A" }, { "family_name": "Milenkovic", "given_name": "Ljiljana", "clpid": "Milenkovic-L" }, { "family_name": "Scott", "given_name": "Matthew P.", "clpid": "Scott-M-P" }, { "family_name": "Davis", "given_name": "Ronald W.", "clpid": "Davis-R-W" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The preimplantation development of the mammalian embryo encompasses a series of critical events: the transition from oocyte to embryo, the first cell divisions, the establishment of cellular contacts, the first lineage differentiation\u2014all the first subtle steps toward a future body plan. Here, we use microarrays to explore gene activity during preimplantation development. We reveal robust and dynamic patterns of stage-specific gene activity that fall into two major phases, one up to the 2-cell stage (oocyte-to-embryo transition) and one after the 4-cell stage (cellular differentiation). The mouse oocyte and early embryo express components of multiple signaling pathways including those downstream of Wnt, BMP, and Notch, indicating that conserved regulators of cell fate and pattern formation are likely to function at the earliest embryonic stages. Overall, these data provide a detailed temporal profile of gene expression that reveals the richness of signaling processes in early mammalian development.", "doi": "10.1016/s1534-5807(03)00404-0", "issn": "1534-5807", "publisher": "Cell Press", "publication": "Developmental Cell", "publication_date": "2004-01", "series_number": "1", "volume": "6", "issue": "1", "pages": "133-144" }, { "id": "authors:8f3rk-h7p71", "collection": "authors", "collection_id": "8f3rk-h7p71", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736360", "type": "book_section", "title": "Cell Lineage Analysis: Applications of Green Fluorescent Protein", "book_title": "Developmental Biology Protocols", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Pines", "given_name": "Jonathon", "orcid": "0000-0002-5227-6004", "clpid": "Pines-J" } ], "contributor": [ { "family_name": "Walker", "given_name": "John M.", "clpid": "Walker-J-M" }, { "family_name": "Tuan", "given_name": "Rocky S.", "clpid": "Tuan-Rocky-S" }, { "family_name": "Lo", "given_name": "Cecilia W.", "clpid": "Lo-Cecilia-W" } ], "abstract": "The ideal cell lineage marker is one that can be visualized in living tissues without perturbing development. Exogenously applied dyes are very useful, but in many instances there would be advantages to an endogenously expressed marker. Green fluorescent protein (GFP) appears to have the potential to be ideal for this purpose, and with its advent, lineage analysis is entering a new phase in which cells can now be followed in real time in living embryos. GFP is a 27-kDa protein found in the jellyfish Aequorea victoria that absorbs blue light and emits green light. It has the valuable property that the formation of the chromophore is an autocatalytic event that requires no cofactors. Furthermore, the cDNA that encodes GFP has been cloned, and thus GFP can be expressed and will become fluorescent in any organism or cell type. In this chapter, we describe the properties of GFP and the different mutants available to study cell lineages in different organisms. We will mainly concentrate on their application to the analysis of cell lineages in frogs and mice.", "doi": "10.1385/1-59259-685-1:279", "isbn": "1592596851", "publisher": "Humana Press", "place_of_publication": "Totowa, NJ", "publication_date": "2003-11-15", "pages": "279-287" }, { "id": "authors:fsy97-3g663", "collection": "authors", "collection_id": "fsy97-3g663", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105738699", "type": "article", "title": "RNA Interference by Production of Short Hairpin dsRNA in ES Cells, Their Differentiated Derivatives, and in Somatic Cell Lines", "author": [ { "family_name": "Grabarek", "given_name": "J. B.", "clpid": "Grabarek-J-B" }, { "family_name": "Wianny", "given_name": "F.", "clpid": "Wianny-F" }, { "family_name": "Plusa", "given_name": "B.", "clpid": "Plusa-B" }, { "family_name": "Zernicka-Goetz", "given_name": "M.", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Glover", "given_name": "D. M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" } ], "abstract": "dsRNA of several hundred nucleotides in length is effective at interfering with gene expression in mouse oocytes, pre-implantation embryos, and embryonic stem (ES) cells but is not as efficient in differentiated cell lines. Here we describe a method to achieve RNA interference in totipotent and differentiated ES cells together with a wide range of other mammalian cell types that is both simple and efficient. It utilizes a linearized plasmid that directs the expression of a hairpin RNA with a 22-nucleotide-paired region. This molecule has a 13-nucleotide 5\u2032 overhang that would be subject to capping on its 5\u2032 phosphoryl group and thus differs from the ideal structure suggested for effective small interfering RNAs. Thus, it appears either that the structure of small inhibitory RNA molecules may not need to be as precise as previously thought or that such a transcript is efficiently processed to a form that is effective in interfering with gene expression.", "doi": "10.2144/03344st02", "issn": "0736-6205", "publisher": "Informa Healthcare", "publication": "BioTechniques", "publication_date": "2003-04", "series_number": "4", "volume": "34", "issue": "4", "pages": "734-744" }, { "id": "authors:76dbt-kr972", "collection": "authors", "collection_id": "76dbt-kr972", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190419-105935323", "type": "article", "title": "Developmental fate of embryonic germ cells (EGCs), in vivo and in vitro", "author": [ { "family_name": "Durcova-Hills", "given_name": "Gabriela", "clpid": "Durcova-Hills-G" }, { "family_name": "Wianny", "given_name": "Florence", "clpid": "Wianny-F" }, { "family_name": "Merriman", "given_name": "Julie", "clpid": "Merriman-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "McLaren", "given_name": "Anne", "clpid": "McLaren-A" } ], "abstract": "Embryonic germ cells (EGCs) derived from mouse primordial germ cells (PGCs) are known both to colonize all cell lineages of the fetus and to make tumors in vivo. When aggregated with eight-cell embryos, EGCs from a new EGC line expressing green fluorescent protein (GFP) were found to contribute preferentially to the epiblast but unexpectedly were also capable of colonizing primary endoderm. When injected under the kidney capsule, EGCs derived from 12.5 days post coitum (dpc) PGCs formed differentiated tumors. The ability of EGCs to differentiate in an organ culture system depends upon their partners in cell culture. When EGCs, marked with a LacZ transgene, were mixed with disaggregated and reaggregated mouse fetal lung in an organ culture system, they remained undifferentiated. In urogenital ridge reaggregates on the other hand, some EGCs were capable of differentiating to form small epithelial cysts.", "doi": "10.1046/j.1432-0436.2003.710204.x", "issn": "0301-4681", "publisher": "Elsevier", "publication": "Differentiation", "publication_date": "2003-03", "series_number": "2", "volume": "71", "issue": "2", "pages": "135-141" }, { "id": "authors:5efyq-h3313", "collection": "authors", "collection_id": "5efyq-h3313", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-100408460", "type": "article", "title": "Determining the first cleavage of the mouse zygote", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "I have been given the opportunity to respond to the article by Gardner and Davies (2003) that appears in this issue criticising some of the experimental methods used by my group to show that the sperm entry position (SEP), when followed either with fluorescent markers attached to the fertilization cone or with labelled spermatozoa, marks the plane of the first cleavage division of the mouse zygote. In their article, Gardner and Davies criticise our studies on two main points: first, that we did not provide sufficient numbers in control groups of eggs, and second, that phytohaemoglutinin (PHA)-coated fluorescent beads, fluorescently-labelled spermatozoa, and fluorescently-labelled concanavalin A (Con A) solution will not mark the SEP properly. Instead, Gardner and Davies maintain that they have performed a better assay. Below, I outline our reasons for believing that these criticisms seem unnecessary. However, in spite of any disagreement about the importance of the sperm entry, it is pleasing that my group and Richard Gardner's do agree that both axes of the blastocyst are already specified before cleavage. Few findings are accepted in science without independent verification and so time will tell which interpretation of the significance of the spermatozoon is closer to the truth.", "doi": "10.1016/s1472-6483(10)61704-x", "issn": "1472-6483", "publisher": "Elsevier", "publication": "Reproductive BioMedicine Online", "publication_date": "2003-01", "series_number": "2", "volume": "6", "issue": "2", "pages": "160-163" }, { "id": "authors:71a38-eqz76", "collection": "authors", "collection_id": "71a38-eqz76", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-100408600", "type": "book_section", "title": "RNA Interference in Mammalian Systems - A Practical Approach", "book_title": "Peroxisomal Disorders and Regulation of Genes", "author": [ { "family_name": "Grabarek", "given_name": "Joanna B.", "clpid": "Grabarek-J-B" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "contributor": [ { "family_name": "Roels", "given_name": "Frank", "clpid": "Roels-F" }, { "family_name": "Baes", "given_name": "Myriam", "clpid": "Baes-M" }, { "family_name": "De Bie", "given_name": "Sylvia", "clpid": "De-Bie-S" } ], "abstract": "RNAi-related mechanisms are evolutionarily conserved and are a tool for cells to defend against foreign nucleic acid invasion and thus genomic instability in the cell. Double-stranded RNA is relatively easy to produce in vitro and, its introduction generates knockout phenotypes without the need to interfere at the DNA level. Thus, the discovery of RNAi has greatly contributed to research by providing a new genetic tool. \n\nThis chapter presents the comparative view on the RNAi-releated techniques and discusses their practical applications in different mammalian experimental systems, including their therapeutic potential.", "doi": "10.1007/978-1-4419-9072-3_24", "isbn": "9781461347828", "publisher": "Springer", "place_of_publication": "Boston, MA", "publication_date": "2003", "pages": "205-216" }, { "id": "authors:4de4w-mwe26", "collection": "authors", "collection_id": "4de4w-mwe26", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-100408698", "type": "article", "title": "Early patterning of the mouse embryo -- contributions of sperm and egg", "author": [ { "family_name": "Piotrowska", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The first cleavage of the fertilised mouse egg divides the zygote into two cells that have a tendency to follow distinguishable fates. One divides first and contributes its progeny predominantly to the embryonic part of the blastocyst, while the other, later dividing cell, contributes mainly to the abembryonic part. We have previously observed that both the plane of this first cleavage and the subsequent order of blastomere division tend to correlate with the position of the fertilisation cone that forms after sperm entry. But does sperm entry contribute to assigning the distinguishable fates to the first two blastomeres or is their fate an intrinsic property of the egg itself? To answer this question we examined the distribution of the progeny of early blastomeres in embryos never penetrated by sperm \u2014 parthenogenetic embryos. In contrast to fertilised eggs, we found there is no tendency for the first two parthenogenetic blastomeres to follow different fates. This outcome is independent of whether parthenogenetic eggs are haploid or diploid. Also unlike fertilised eggs, the first 2-cell blastomere to divide in parthenogenetic embryo does not necessarily contribute more cells to the blastocyst. However, even when descendants of the first dividing blastomere do predominate, they show no strong predisposition to occupy the embryonic part. Thus blastomere fate does not appear to be decided by differential cell division alone. Finally, when the cortical cytoplasm at the site of sperm entry is removed, the first cleavage plane no longer tends to divide the embryo into embryonic and abembryonic parts. Together these results indicate that in normal development fertilisation contributes to setting up embryonic patterning, alongside the role of the egg.", "doi": "10.1242/dev.00170", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "2002-12-15", "series_number": "24", "volume": "129", "issue": "24", "pages": "5803-5813" }, { "id": "authors:7x20y-hr337", "collection": "authors", "collection_id": "7x20y-hr337", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-100408799", "type": "article", "title": "Site of the previous meiotic division defines cleavage orientation in the mouse embryo", "author": [ { "family_name": "Plusa", "given_name": "Berenika", "clpid": "Plusa-B" }, { "family_name": "Grabarek", "given_name": "Joanna B.", "clpid": "Grabarek-J-B" }, { "family_name": "Piotrowska", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The conservation of early cleavage patterns in organisms as diverse as echinoderms and mammals suggests that even in highly regulative embryos such as the mouse, division patterns might be important for development. Indeed, the first cleavage divides the fertilized mouse egg into two cells: one cell that contributes predominantly to the embryonic part of the blastocyst, and one that contributes to the abembryonic part. Here we show, by removing, transplanting or duplicating the animal or vegetal poles of the mouse egg, that a spatial cue at the animal pole orients the plane of this initial division. Embryos with duplicated animal, but not vegetal, poles show abnormalities in chromosome segregation that compromise their development. Our results show that localized factors in the mammalian egg orient the spindle and so define the initial cleavage plane. In increased dosage, however, these factors are detrimental to the correct execution of division.", "doi": "10.1038/ncb860", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2002-10", "series_number": "10", "volume": "4", "issue": "10", "pages": "811-815" }, { "id": "authors:3g87x-0bc76", "collection": "authors", "collection_id": "3g87x-0bc76", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-100408907", "type": "article", "title": "Efficient delivery of dsRNA into zona-enclosed mouse oocytes and preimplantation embryos by electroporation", "author": [ { "family_name": "Grabarek", "given_name": "Joanna B.", "clpid": "Grabarek-J-B" }, { "family_name": "Plusa", "given_name": "Berenika", "clpid": "Plusa-B" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Conditions for the electroporation of mouse oocytes and preimplantation embryos have been optimised by following the incorporation of rhodamine labeled dextran. This procedure includes a step to weaken but not remove the zona pellucida that helps achieve good survival. This approach has been applied to introduce double\u2010stranded RNA for c\u2010mos into oocytes and green fluorescent protein (GFP) into transgenic GFP\u2010expressing embryos at the 1\u2010 and 4\u2010cell stages. In both cases we were able to observe sequence\u2010specific interference with the expression of the target gene\u2014a failure of oocytes to arrest at metaphase II and a loss in the green fluorescence of embryos by the morula or blastocyst stages. These effects could be observed in multiple oocytes or embryos allowed to develop together following electroporation.", "doi": "10.1002/gene.10076", "issn": "1526-954X", "publisher": "Wiley", "publication": "Genesis", "publication_date": "2002-04", "series_number": "4", "volume": "32", "issue": "4", "pages": "269-276" }, { "id": "authors:k91nf-g5a80", "collection": "authors", "collection_id": "k91nf-g5a80", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-100408995", "type": "article", "title": "Sperm entry position provides a surface marker for the first cleavage plane of the mouse zygote", "author": [ { "family_name": "Plusa", "given_name": "Berenika", "clpid": "Plusa-B" }, { "family_name": "Piotrowska", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The sperm entry position (SEP) of the mouse egg, labelled by placing a bead at the fertilisation cone, tends to be associated with the first cleavage plane (Piotrowska and Zernicka\u2010Goetz: Nature 409:517\u2013521, 2001). Nevertheless, in up to one\u2010fourth of embryos the cleavage furrow did not pass close to the bead, and following the division the bead marked the cleavage plane in only 60% of cases. This raised the question of whether such variability arose from the labelling itself or had a biological basis. The zona pellucida was not responsible for this effect because similar results were obtained in its presence or absence. However, this variability could be attributable to the large size of the fertilisation cone relative to the SEP. Therefore, we have developed a means of fluorescently labelling sperm that can record the exact site of its penetration when the label transfers to the egg surface. This approach indicates that the SEP marks the first cleavage in the great majority (88%) of embryos. In conclusion, direct sperm labelling shows precisely the correlation between the SEP and the first cleavage, although there is natural variability in this process.", "doi": "10.1002/gene.10027", "issn": "1526-954X", "publisher": "Wiley", "publication": "Genesis", "publication_date": "2002-03", "series_number": "3", "volume": "32", "issue": "3", "pages": "193-198" }, { "id": "authors:7w1qa-xgr16", "collection": "authors", "collection_id": "7w1qa-xgr16", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-100425160", "type": "article", "title": "Patterning of the embryo: the first spatial decisions in the life of a mouse", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Although in most species the polarity of the embryo takes its roots from the spatial patterning of the egg, mammals were viewed as an exception. This was because the anteroposterior polarity of the mouse embryo could not be seen until gastrulation, and no developmental cues were known that could define polarity at earlier stages. Why should we now re-consider this view? While mechanisms of axis formation in mammals could, in principle, be unique, the evolutionary conservation of numerous other developmental processes raises the question of why mammals would have evolved a different way or timing of organising their embryonic polarity. Indeed, recent evidence shows that well before the onset of gastrulation, the mouse embryo initiates asymmetric patterns of gene expression in its visceral endoderm. Although this extra-embryonic tissue does not contribute to the body itself, it is involved in axis formation. Other recent work has revealed that spatial distribution of cells in the visceral endoderm can be traced back to polarity present at the blastocyst stage. These insights have raised the possibility that embryonic polarity might also originate early during development of mammalian embryos. Indeed it now appears that there are at least two spatial cues that operate in the mouse egg to shape polarity of the blastocyst. One of these is at the animal pole, which is defined by the site of female meiosis, and another is associated with the position of sperm entry. In this review I discuss these recent findings, which have led to the recognition that mouse embryos initiate development of their polarity at the earliest stages of their life. This novel perspective raises questions about the nature of cellular and molecular mechanisms that could convert developmental cues in the zygote to axes of the blastocyst, and hence into polarity of the post-implantation embryo. It also brings to light the need to understand how such mechanisms could enable early mouse development to be so regulative.", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "2002-02-15", "series_number": "4", "volume": "129", "issue": "4", "pages": "815-829" }, { "id": "authors:st565-ngg54", "collection": "authors", "collection_id": "st565-ngg54", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-100452455", "type": "article", "title": "Blastomeres arising from the first cleavage division have distinguishable fates in normal mouse development", "author": [ { "family_name": "Piotrowska", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Wianny", "given_name": "Florence", "clpid": "Wianny-F" }, { "family_name": "Pedersen", "given_name": "Roger A.", "clpid": "Pedersen-R-A" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Two independent studies have recently suggested similar models in which the embryonic and abembryonic parts of the mouse blastocyst become separated already by the first cleavage division. However, no lineage tracing studies carried out so far on early embryos provide the support for such a hypothesis. Thus, to re-examine the fate of blastomeres of the two-cell mouse embryo, we have undertaken lineage tracing studies using a non-perturbing method. We show that two-cell stage blastomeres have a strong tendency to develop into cells that comprise either the embryonic or the abembryonic parts of the blastocyst. Moreover, the two-cell stage blastomere that is first to divide will preferentially contribute its progeny to the embryonic part. Nevertheless, we find that the blastocyst embryonic-abembryonic axis is not perfectly orthogonal to the first cleavage plane, but often shows some angular displacement from it. Consequently, there is a boundary zone adjacent to the interior margin of the blastocoel that is populated by cells derived from both earlier and later dividing blastomeres. The majority of cells that inhabit this boundary region are, however, derived from the later dividing two-cell stage blastomere that contributes predominantly to the abembryonic part of the blastocyst. Thus, at the two-cell stage it is already possible to predict which cell will contribute a greater proportion of its progeny to the abembryonic part of the blastocyst (region including the blastocyst cavity) and which to the embryonic part (region containing the inner cell mass) that will give rise to the embryo proper.", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "2001-10", "series_number": "19", "volume": "128", "issue": "19", "pages": "3739-3748" }, { "id": "authors:s3qda-39884", "collection": "authors", "collection_id": "s3qda-39884", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101735870", "type": "article", "title": "Use of Green Fluorescent Protein in Mouse Embryos", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Pines", "given_name": "Jonathon", "orcid": "0000-0002-5227-6004", "clpid": "Pines-J" } ], "abstract": "Green Fluorescent Protein (GFP) has rapidly been established as a versatile and powerful cell marker in many organisms. Initial problems in using it in mammalian cells were solved by introducing mutations to increase its solubility at higher temperatures, such that GFP has now been used as a reporter in both gene expression and cell lineage studies, and to localize proteins within mammalian cells. GFP has two unique advantages: (i) the protein becomes fluorescent in an autocatalytic reaction, so that it can be introduced into any cell type simply as a cDNA or mRNA, or as protein; (ii) it is \"bright\" enough to be visualized in living cells under conditions that do not cause photodamage to the cells. In this article we outline the ways in which we have used GFP mRNA and cDNA in our studies of mouse cell lineages, and to characterize the behavior of proteins within the embryos.", "doi": "10.1006/meth.2001.1157", "issn": "1046-2023", "publisher": "Elsevier", "publication": "Methods", "publication_date": "2001-05", "series_number": "1", "volume": "24", "issue": "1", "pages": "55-60" }, { "id": "authors:svjpx-6mz35", "collection": "authors", "collection_id": "svjpx-6mz35", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101735996", "type": "article", "title": "Role for sperm in spatial patterning of the early mouse embryo", "author": [ { "family_name": "Piotrowska", "given_name": "Karolina", "clpid": "Piotrowska-Nitsche-K" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Despite an apparent lack of determinants that specify cell fate, spatial patterning of the mouse embryo is evident early in development. The axis of the post-implantation egg cylinder can be traced back to organization of the pre-implantation blastocyst. This in turn reflects the organization of the cleavage-stage embryo and the animal\u2013vegetal axis of the zygote. These findings suggest that the cleavage pattern of normal development may be involved in specifying the future embryonic axis; however, how and when this pattern becomes established is unclear. In many animal eggs, the sperm entry position provides a cue for embryonic patterning, but until now no such role has been found in mammals. Here we show that the sperm entry position predicts the plane of initial cleavage of the mouse egg and can define embryonic and abembryonic halves of the future blastocyst. In addition, the cell inheriting the sperm entry position acquires a division advantage and tends to cleave ahead of its sister. As cell identity reflects the timing of the early cleavages, these events together shape the blastocyst whose organization will become translated into axial patterning after implantation. We present a model for axial development that accommodates these findings with the regulative nature of mouse embryos.", "doi": "10.1038/35054069", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2001-01-25", "series_number": "6819", "volume": "409", "issue": "6819", "pages": "517-521" }, { "id": "authors:v3jkt-qp874", "collection": "authors", "collection_id": "v3jkt-qp874", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101707437", "type": "article", "title": "Animal and vegetal poles of the mouse egg predict the polarity of the embryonic axis, yet are nonessential for development", "author": [ { "family_name": "Ciemerych", "given_name": "Maria Anna", "clpid": "Ciemerych-M-A" }, { "family_name": "Mesnard", "given_name": "Daniel", "clpid": "Mesnard-D" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Recent studies suggest early (preimplantation) events might be important in the development of polarity in mammalian embryos. We report here lineage tracing experiments with green fluorescent protein showing that cells located either near to or opposite the polar body at the 8-cell stage of the mouse embryo retain their same relative positions in the blastocyst. Thus they come to lie on either end of an axis of symmetry of the blastocyst that has recently been shown to correlate with the anterior-posterior axis of the postimplantation embryo (see R. J. Weber, R. A. Pedersen, F. Wianny, M. J. Evans and M. Zernicka-Goetz (1999). Development 126, 5591-5598). The embryonic axes of the mouse can therefore be related to the position of the polar body at the 8-cell stage, and by implication, to the animal-vegetal axis of the zygote. However, we also show that chimeric embryos constructed from 2-cell stage blastomeres from which the animal or the vegetal poles have been removed can develop into normal blastocysts and become fertile adult mice. This is also true of chimeras composed of animal or vegetal pole cells derived through normal cleavage to the 8-cell stage. We discuss that although polarity of the postimplantation embryo can be traced back to the 8-cell stage and in turn to the organisation of the egg, it is not absolutely fixed by this time.", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "2000-08-15", "series_number": "16", "volume": "127", "issue": "16", "pages": "3467-3474" }, { "id": "authors:sgbph-qjh48", "collection": "authors", "collection_id": "sgbph-qjh48", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736085", "type": "article", "title": "Jumping the gun on mouse gene expression", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "I am writing to correct a statement in the informative News Feature \"A silence that speaks volumes\" (Nature 404, 804; 2000). The work from my laboratory, published in the paper by Florence Wianny and myself, has shown that dsRNA interference can be effective in both the mouse oocyte and early embryo (see Wianny, F. & Zernicka-Goetz, M. Nature Cell Biol. 2, 70\u201375; 2000). We have successfully used the technique to prevent expression of the c-mos gene during oocyte maturation, and of the E-cadherin gene to interfere with formation of the blastocyst when delivered to the one-cell-stage zygote.", "doi": "10.1038/35015787", "issn": "0028-0836", "publisher": "Nature Publishing Group", "publication": "Nature", "publication_date": "2000-06-15", "series_number": "6788", "volume": "405", "issue": "6788", "pages": "733" }, { "id": "authors:sw9xb-g4k94", "collection": "authors", "collection_id": "sw9xb-g4k94", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736181", "type": "article", "title": "Progression of mouse oocytes from metaphase I to metaphase II is inhibited by fusion with G2 cells", "author": [ { "family_name": "Grabarek", "given_name": "Joanna", "clpid": "Grabarek-J" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "We show that in contrast to metaphase II oocytes, metaphase I oocytes cannot be activated by fusion with the zygote. Fusion of metaphase I oocytes with G2 zygotes was followed by premature chromosome condensation, with 60% of the hybrids becoming arrested at metaphase I, the remainder progressing and arresting at metaphase II. Hybrids of metaphase I oocytes and M-phase zygotes underwent accelerated maturation, but all arrested at metaphase II. In both cases the arrest could be overcome by treatment with the parthenogenetic activators ethanol and cycloheximide. We discuss these findings in relation to the possibility that the metaphase I oocyte contains cytostatic factor activity that is activated by its zygotic partner. Alternatively, the G2 zygote may provide an inhibitor of anaphase, normally never present in the metaphase I oocyte and which is absent from the M-phase zygote.", "doi": "10.1017/s0967199400000927", "issn": "0967-1994", "publisher": "Cambridge University Press", "publication": "Zygote", "publication_date": "2000-05", "series_number": "2", "volume": "8", "issue": "2", "pages": "145-151" }, { "id": "authors:1ja12-3x079", "collection": "authors", "collection_id": "1ja12-3x079", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736270", "type": "article", "title": "Specific interference with gene function by double-stranded RNA in early mouse development", "author": [ { "family_name": "Wianny", "given_name": "Florence", "clpid": "Wianny-F" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "The use of double-stranded (ds) RNA is a powerful way of interfering with gene expression in a range of organisms, but doubts have been raised about whether it could be successful in mammals. Here, we show that dsRNA is effective as a specific inhibitor of the function of three genes in the mouse, namely maternally expressed c-mos in the oocyte and zygotically expressed E-cadherin or a GFP transgene in the preimplantation embryo. The phenotypes observed are the same as those reported for null mutants of the endogenous genes. These findings offer the opportunity to study development and gene regulation in normal and diseased cells.", "doi": "10.1038/35000016", "issn": "1465-7392", "publisher": "Nature Publishing Group", "publication": "Nature Cell Biology", "publication_date": "2000-02", "series_number": "2", "volume": "2", "issue": "2", "pages": "70-75" }, { "id": "authors:2jqnk-kdv96", "collection": "authors", "collection_id": "2jqnk-kdv96", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101708618", "type": "article", "title": "Polarity of the mouse embryo is anticipated before implantation", "author": [ { "family_name": "Weber", "given_name": "R. J.", "clpid": "Weber-R-J" }, { "family_name": "Pedersen", "given_name": "R. A.", "clpid": "Pedersen-R-A" }, { "family_name": "Wianny", "given_name": "F.", "clpid": "Wianny-F" }, { "family_name": "Evans", "given_name": "M. J.", "clpid": "Evans-M-J-Bio" }, { "family_name": "Zernicka-Goetz", "given_name": "M.", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "In most species, the polarity of an embryo underlies the future body plan and is determined from that of the zygote. However, mammals are thought to be an exception to this; in the mouse, polarity is generally thought to develop significantly later, only after implantation. It has not been possible, however, to relate the polarity of the preimplantation mouse embryo to that of the later conceptus due to the lack of markers that endure long enough to follow lineages through implantation. To test whether early developmental events could provide cues that predict the axes of the postimplantation embryo, we have used the strategy of injecting mRNA encoding an enduring marker to trace the progeny of inner cell mass cells into the postimplantation visceral endoderm. This tissue, although it has an extraembryonic fate, plays a role in axis determination in adjacent embryonic tissue. We found that visceral endoderm cells that originated near the polar body (a marker of the blastocyst axis of symmetry) generally became distal as the egg cylinder formed, while those that originated opposite the polar body tended to become proximal. It follows that, in normal development, bilateral symmetry of the mouse blastocyst anticipates the polarity of the later conceptus. Moreover, our results show that transformation of the blastocyst axis of symmetry into the axes of the postimplantation conceptus involves asymmetric visceral endoderm cell movement. Therefore, even if the definitive axes of the mouse embryo become irreversibly established only after implantation, this polarity can be traced back to events before implantation.", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "1999-12-15", "series_number": "24", "volume": "126", "issue": "24", "pages": "5591-5598" }, { "id": "authors:9ycvq-k2r52", "collection": "authors", "collection_id": "9ycvq-k2r52", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101709593", "type": "article", "title": "Fertile offspring derived from mammalian eggs lacking either animal or vegetal poles", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "In all animals so far tested, removing either pole of the undivided egg prevents normal development: embryos may arrest early, lack organs, or the adults may be sterile. These experiments have shown that spatial patterning of the egg is of utmost importance for subsequent development. However, the significance of spatial patterning in mammalian eggs is still controversial. To test the importance of egg polarity in the mouse a substantial amount of material either from the animal (polar body-associated) or the vegetal (opposite) pole of the fertilised egg was removed. One pole of the egg was cut away manually with a glass needle and the eggs were allowed to develop in vitro. Both kinds of surgical operation permit the development of blastocysts, which, after transfer to the uteri of pseudo-pregnant foster mothers, can produce viable offspring. Furthermore, these develop into fertile adult mice. I conclude that mouse eggs have no essential components that are localised uniquely to the animal or the vegetal pole and, therefore, do not rely for their axial development on maternal determinants that are so localised in the fertilised egg. Thus the mammalian egg appears to be very unusual in the animal kingdom in that it establishes the embryonic axes after the zygote has begun development.", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "1998-12", "series_number": "23", "volume": "125", "issue": "23", "pages": "4803-4808" }, { "id": "authors:ge56g-h5h39", "collection": "authors", "collection_id": "ge56g-h5h39", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736448", "type": "article", "title": "Mouse polo-like kinase 1 associates with the acentriolar spindle poles, meiotic chromosomes and spindle midzone during oocyte maturation", "author": [ { "family_name": "Wianny", "given_name": "Florence", "clpid": "Wianny-F" }, { "family_name": "Tavares", "given_name": "\u00c1lvaro", "clpid": "Tavares-\u00c1" }, { "family_name": "Evans", "given_name": "Martin J.", "clpid": "Eans-M-J" }, { "family_name": "Glover", "given_name": "David M.", "orcid": "0000-0003-0956-0103", "clpid": "Glover-D-M" }, { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "We have examined the dynamics of the localisation of the polo-like kinase 1 (Plk1) during maturation of the mouse oocyte. Levels of Plk1 protein increase following germinal vesicle breakdown, at which time the enzyme begins to accumulate at discrete positions on the condensing chromosomes and, subsequently, at the poles of the meiotic spindle, which moves towards the cortex of the egg. Interestingly, at metaphase in both meiotic divisions, Plk1 shows a punctate localisation along the broad spindle poles. Moreover, the punctate distribution of Plk1 on the meiotic chromosomes appears at early anaphase to correspond to the centromeric regions. The protein relocates to the spindle midzone during late anaphase and then associates with the midbody at telophase. We have confirmed the specific pattern of immuno-localisation seen in fixed preparations by observing the distribution of Plk1 tagged with green fluorescent protein in living oocytes. We discuss the localisation of the enzyme in light of the structure of the spindle poles, which are known to lack centrioles, and the highly asymmetric nature of the meiotic divisions.", "doi": "10.1007/s004120050327", "issn": "0009-5915", "publisher": "Springer", "publication": "Chromosoma", "publication_date": "1998-12", "series_number": "6-7", "volume": "107", "issue": "6-7", "pages": "430-439" }, { "id": "authors:88m99-c2788", "collection": "authors", "collection_id": "88m99-c2788", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101710597", "type": "article", "title": "Following cell fate in the living mouse embryo", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "M.", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Pines", "given_name": "J.", "orcid": "0000-0002-5227-6004", "clpid": "Pines-J" }, { "family_name": "McLean Hunter", "given_name": "S.", "clpid": "McLean-Hunter-S" }, { "family_name": "Dixon", "given_name": "J. P. C.", "clpid": "Dixon-J-P-C" }, { "family_name": "Siemering", "given_name": "K. R.", "clpid": "Siemering-K-R" }, { "family_name": "Haseloff", "given_name": "J.", "clpid": "Haseloff-J" }, { "family_name": "Evans", "given_name": "M. J.", "clpid": "Evans-M-J-Bio" } ], "abstract": "It has been difficult to follow many of the dramatic changes in cell fate and cell migration during mouse development. This is because there has been no enduring marker that would allow cells to be recognised in the living embryo. We believe that we have overcome this problem by developing a novel form of green fluorescent protein, named MmGFP, that proves to be easily visible and non toxic to mouse cells and does not perturb embryogenesis. We show that synthetic mRNA encoding MmGFP can be injected into blastomeres to follow the fate of their progeny during preimplantation development. We have made a stable embryonic stem cell line that expresses MmGFP and introduced these fluorescent cells into mouse embryos. For the first time, we have been able to follow the fate of embryonic stem cells in living embryos and to observe directly the contribution of these cells to distinct lineages of the postimplantation embryo. This approach should lead to a more complete description of the dynamics of cell fate in the mouse.", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "1997-03-15", "series_number": "6", "volume": "124", "issue": "6", "pages": "1133-1137" }, { "id": "authors:1ssna-chh64", "collection": "authors", "collection_id": "1ssna-chh64", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101711630", "type": "article", "title": "Protein phosphatases control MAP kinase activation and microtubule organization during rat oocyte maturation", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "M.", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Verlhac", "given_name": "M. H.", "clpid": "Verlhac-M-H" }, { "family_name": "G\u00e9raud", "given_name": "G.", "clpid": "G\u00e9raud-G" }, { "family_name": "Kubiak", "given_name": "J. Z.", "clpid": "Kubiak-J-Z" } ], "abstract": "Mitogen-activated protein kinase (MAP) is involved in many signal transduction pathways and is activated during meiotic maturation in various species. In this study, we used the rat oocyte to identify some of the control mechanisms involved in MAP kinase activation which is triggered at resumption of meiosis. We examined the respective contribution of this kinase and maturation promoting factor (MPF), or cdc2 kinase, in the regulation of microtubule behavior and in the reorganization of chromatin during meiotic maturation. We found that the resumption of meiotic division in rat oocytes coincided with the activation of MPF and was followed 3 h later by the activation of MAP kinase. The activation of the two kinases also occurred in oocytes undergoing maturation in the presence of the protein phosphatase inhibitor okadaic acid (OA). However, the activation of cdc2 kinase was only partial, whereas activation of MAP kinase was accelerated and began 1 h after the resumption of meiosis, i.e. 2 h earlier than in control oocytes. We also showed that protein synthesis was required to activate MAP kinase, but not cdc2 kinase. However, once MAP kinase was activated, ongoing protein synthesis was not necessary to maintain its activity. These results suggest that a negative regulation of MAP kinase slows down its activation at the resumption of meiosis, mediated through the level of phosphatase activity. Moreover, MAP kinase activation requires protein synthesis, even upon phosphatase inactivation by OA, suggesting also the existence of a positive control pathway. We observed that during the first meiotic M-phase, the spindle did not form immediately after cdc2 kinase activation, but that its formation coincided with the appearance of MAP kinase activity. However, earlier activation of MAP kinase by treatment with OA did not lead to premature spindle formation, but instead a large aster formed consisting of long microtubules radiating from the condensed chromatin. In OA-treated oocytes, spindles did not form and an interphase network of microtubule developed with time. Thus, MAP kinase is unable to substitute for MPF under these conditions, its activity alone being insufficient to maintain the progression through meiotic maturation.", "issn": "0171-9335", "publisher": "Elsevier", "publication": "European journal of cell biology", "publication_date": "1997-01", "series_number": "1", "volume": "72", "issue": "1", "pages": "30-38" }, { "id": "authors:8byqp-ahs04", "collection": "authors", "collection_id": "8byqp-ahs04", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101712629", "type": "article", "title": "An indelible lineage marker for Xenopus using a mutated green fluorescent protein", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "M.", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Pines", "given_name": "J.", "orcid": "0000-0002-5227-6004", "clpid": "Pines-J" }, { "family_name": "Ryan", "given_name": "K.", "clpid": "Ryan-K" }, { "family_name": "Siemering", "given_name": "K. R.", "clpid": "Siemering-K-R" }, { "family_name": "Haseloff", "given_name": "J.", "clpid": "Haseloff-J" }, { "family_name": "Evans", "given_name": "M. J.", "clpid": "Evans-M-J-Bio" }, { "family_name": "Gurdon", "given_name": "J. B.", "clpid": "Gurdon-J-B" } ], "abstract": "We describe the use of a DNA construct (named GFP.RN3) encoding green fluorescent protein as a lineage marker for Xenopus embryos. This offers the following advantages over other lineage markers so far used in Xenopus. When injected as synthetic mRNA, its protein emits intense fluorescence in living embryos. It is non-toxic, and the fluorescence does not bleach when viewed under 480 nm light. It is surprisingly stable, being strongly visible up to the feeding tadpole stage (5 days), and in some tissues for several weeks after mRNA injection. We also describe a construct that encodes a blue fluorescent protein. We exemplify the use of this GFP.RN3 construct for marking the lineage of individual blastomeres at the 32- to 64-cell stage, and as a marker for single transplanted blastula cells. Both procedures have revealed that the descendants of one embryonic cell can contribute single muscle cells to nearly all segmental myotomes rather than predominantly to any one myotome. An independent aim of our work has been to follow the fate of cells in which an early regulatory gene has been temporarily overexpressed. For this purpose, we co-injected GFP.RN3 mRNA and mRNA for the early Xenopus gene Eomes, and found that a high concentration of Eomes results in ectopic muscle gene activation in only the injected cells. This marker may therefore be of general value in providing long term identification of those cells in which an early gene with ephemeral expression has been overexpressed.", "issn": "0950-1991", "publisher": "Company of Biologists", "publication": "Development", "publication_date": "1996-12", "series_number": "12", "volume": "122", "issue": "12", "pages": "3719-3724" }, { "id": "authors:kdfws-e3m86", "collection": "authors", "collection_id": "kdfws-e3m86", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101713745", "type": "article", "title": "Cytostatic factor inactivation is induced by a calcium-dependent mechanism present until the second cell cycle in fertilized but not in parthenogenetically activated mouse eggs", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Ciemerych", "given_name": "Maria A.", "clpid": "Ciemerych-M-A" }, { "family_name": "Kubiak", "given_name": "Jacek Z.", "clpid": "Kubiak-J-Z" }, { "family_name": "Tarkowski", "given_name": "Andrzej K.", "clpid": "Tarkowski-A-K" }, { "family_name": "Maro", "given_name": "Bernard", "clpid": "Maro-B" } ], "abstract": "Cytostatic factor (CSF) is an activity responsible for the metaphase II arrest in vertebrate oocytes. This activity maintains a high level of maturation promoting factor (MPF) in the oocyte and both activities are destroyed after fertilization or parthenogenetic activation. To study some of the characteristics of the mechanism involved in MPF and CSF destruction, we constructed hybrid cells between metaphase II arrested oocytes and early embryos obtained after fertilization or artificial activation. We found that the behavior of hybrid cells differed depending upon the type of oocyte activation. Initially, the reaction of both types of hybrid cells was similar, the nuclear envelope broke down and chromatin condensation was induced. However, while metaphase II oocytes fused with parthenogenetic eggs remained arrested in M-phase, the oocytes fused with fertilized eggs underwent activation and passed into interphase. This ability of fertilized eggs to induce oocyte activation was still present at the beginning, but not at the end of the second embryonic cell cycle. Oocyte activation induced by fusion with a fertilized egg could be prevented when calcium was chelated by BAPTA. Thus, element(s) of the mechanism involved in calcium release triggered by a sperm component at fertilization remain(s) active until the second cell cycle and is (are) inactivated before the end of the 2-cell stage.", "issn": "0021-9533", "publisher": "Company of Biologists", "publication": "Journal of Cell Science", "publication_date": "1995-02", "series_number": "2", "volume": "108", "issue": "2", "pages": "469-474" }, { "id": "authors:ybvna-rd425", "collection": "authors", "collection_id": "ybvna-rd425", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736535", "type": "article", "title": "Activation of embryonic genes during preimplantation rat development", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Expression of the embryonic genome has been examined during preimplantation rat development. Proteins synthesized at different stages of embryogenesis were labelled with [^(35)S]methionine and then separated by one\u2010dimensional gel electrophoresis. A major transformation in the pattern of protein synthesis has been observed between the two\u2010 and the four\u2010cell stages of embryonic development. Also the culture of embryos with an inhibitor of transcription (\u03b1\u2010amanitin) has shown that the first \u03b1\u2010amanitin\u2010sensitive events take place during the late two\u2010cell stage. However, inhibition of transcription does not arrest the embryo development up to the four\u2010cell stage. Taken together, the results indicate that in rats the initiation of embryonic gene activation occurs at the late two\u2010cell stage. However, the first two cleavage divisions can occur in the absence of transcription.", "doi": "10.1002/mrd.1080380106", "issn": "1040-452X", "publisher": "Wiley-Liss", "publication": "Molecular Reproduction and Development", "publication_date": "1994-05", "series_number": "1", "volume": "38", "issue": "1", "pages": "30-35" }, { "id": "authors:fvsa3-dda03", "collection": "authors", "collection_id": "fvsa3-dda03", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736624", "type": "article", "title": "Okadaic Acid Affects Spindle Organization in Metaphase II-Arrested Rat Oocytes", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Maro", "given_name": "Bernard", "clpid": "Maro-B" } ], "abstract": "We used okadaic acid (OA), a potent phosphatase inhibitor, to study the role of phosphatases in the organization of the spindle microtubules in metaphase II-arrested rat oocytes. OA induced a dramatic elongation of the spindle and disorganization of the metaphase plate. Biochemical analysis revealed some dramatic changes after OA treatment: increased histone H1 kinase activity, a burst of protein phosphorylation, and inhibition of protein synthesis. The latter effect alone cannot be responsible for the changes in the spindle structure since inhibition of protein synthesis with puromycin had no such effect. To identify the targets of OA among the proteins associated with the spindle, meiotic spindles of OA-treated and control oocytes were purified. After OA treatment some proteins gained and others lost their association with the spindle, A few proteins that lost their affinity for the spindle also underwent hyperphosphorylation after OA treatment. These results suggest that hyperphosphorylation of some proteins associated with the spindle alters their affinity to microtubules and thus stabilizes them.", "doi": "10.1006/excr.1993.1179", "issn": "0014-4827", "publisher": "Elsevier", "publication": "Experimental Cell Research", "publication_date": "1993-07", "series_number": "1", "volume": "207", "issue": "1", "pages": "189-193" }, { "id": "authors:5s53s-xz364", "collection": "authors", "collection_id": "5s53s-xz364", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101714782", "type": "article", "title": "Full activation of the rat oocyte by protein synthesis inhibition requires protein phosphatase activity", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Weber", "given_name": "Mich\u00e8le", "clpid": "Weber-M" }, { "family_name": "Maro", "given_name": "Bernard", "clpid": "Maro-B" } ], "abstract": "The rat oocyte provides an interesting system in which to dissect the control mechanisms involved in the transition between a meiotic M phase and a mitotic interphase. In this study, we show that in rat oocytes activated parthenogenetically by puromycin, okadaic acid (a potent inhibitor of protein phosphatases 1 and 2A) induced an increase in histone H1 kinase activity suggesting that MPF was reactivated. However, the inhibition of phosphatases 1 and 2A shortly after second polar body extrusion did not allow the formation of a metaphase-like spindle, although microtubule polymerization was not inhibited. Instead, the chromatin remained condensed as a single mass and a large aster formed around it.", "issn": "0214-6282", "publisher": "University of the Basque Country Press", "publication": "International Journal of Developmental Biology", "publication_date": "1993-06", "series_number": "2", "volume": "37", "issue": "2", "pages": "273-277" }, { "id": "authors:h2b2n-gzn68", "collection": "authors", "collection_id": "h2b2n-gzn68", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736718", "type": "article", "title": "Cytoskeletal organization of rat oocytes during metaphase II arrest and following abortive activation: A study by confocal laser scanning microscopy", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" }, { "family_name": "Kubiak", "given_name": "Jacek Z.", "clpid": "Kubiak-J-Z" }, { "family_name": "Antony", "given_name": "Claude", "clpid": "Antony-C" }, { "family_name": "Maro", "given_name": "Bernard", "clpid": "Maro-B" } ], "abstract": "In metaphase II arrested rat oocytes (M il), microtubles were found in the taper\u2010shaped meiotic spindle and in the cytoplasm as asters and free microtubules. Whereas spindle microtubules were acetylated, those located in the cytoplasm were not. Cytoplasmic microtubules were also labile as assessed by mild cooling. In contast to mouse oocytes, rat microtubule organizing centers (MTOCs) did not react with MPM\u20102 antibody by immunofluorescence despite the fact that this antibody reacts with several proteins as shown by immunoblot. However, cytoplasmic MTOCs in M II\u2010arrested rat oocytes could be detected by their nucleating capacity in the presence of taxol, a drug that induced the formation of numerous cytoplasmic asters. In addition, taxol caused a change in the spindle shape and the formation of astral microtubules at the spindle poles. Meiotic spindles (as well as chromosomes devoid of microtubules after nocodazoletreatment) were overlaid by an actin\u2010rich domain. Spontaneous abortive activation led to the extrusion of the second polar body followed by another metaphase arrest\u2014 metaphase III; however, normal spindles did not form and dispersed chromosomes surrounded by microtubles were observed. Electron microscopic studies confirmed these observations and revealed that the kinetochores are located deep within the chromosomes in contrast to mouse kinetochores, and this might be responsible for the absence of a metaphase III spindle in the rat oocyte. Induced activation caused transition to interphase with the formation of a characteristic microtubule network. This study shows that there are several significant differences in the cytoskeletal organization of rat and mouse oocytes.", "doi": "10.1002/mrd.1080350210", "issn": "1040-452X", "publisher": "Wiley-Liss", "publication": "Molecular Reproduction and Development", "publication_date": "1993-06", "series_number": "2", "volume": "35", "issue": "2", "pages": "165-175" }, { "id": "authors:m1t1b-th494", "collection": "authors", "collection_id": "m1t1b-th494", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190422-101736808", "type": "article", "title": "Spontaneous and induced activation of rat oocytes", "author": [ { "family_name": "Zernicka-Goetz", "given_name": "Magdalena", "orcid": "0000-0002-7004-2471", "clpid": "Zernicka-Goetz-M" } ], "abstract": "Ovulated rat oocytes undergo spontaneous activation during in vitro culture. After extrusion of the second polar body, they do not enter interphase but are arrested again in next metaphase\u2010like stage (M III arrest). The present study demonstrates that puromycin and chloral hydrate can trigger transition to interphase of metaphase II and spontaneously (incompletely) activated rat oocytes. The response of oocytes to these activators depends on their stage at the time of application of a stimulus. Metaphase II oocytes enter interphase at 86.8% when treated with puromycin and in 28.7% after chloral hydrate activation. Oocytes activated with chloral hydrate at the time of spontaneously induced anaphase II enter interphase at 64.8%, but after reaching the stage of telophase II their capability to shift to interphase is again low (28.8%). Finally, M III oocytes cannot be forced to enter interphase by either chloral hydrate or puromycin treatment. This study shows that resumption of the second meiotic division and transition to interphase\u2013\u2010the two processes that normally occur in succession as a response to oocyte activatin\u2010\u2013can be experimentally separated.", "doi": "10.1002/mrd.1080280210", "issn": "1040-452X", "publisher": "Wiley-Liss", "publication": "Molecular Reproduction and Development", "publication_date": "1991-02", "series_number": "2", "volume": "28", "issue": "2", "pages": "169-176" } ]