Morphogenetic ovarian modules for oocyte production from germline stem cells

Morphogenetic ovarian modules for oocyte production from germline stem cells | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Biological Sciences - Article Morphogenetic ovarian modules for oocyte production from germline stem cells Yaniv Elkouby, Neta Hart, Johanna Kroll, Vineet Kumar, Sylwia Czukiewska, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7325126/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Oocyte production from early progenitors in the fetal ovary determines the ovarian reserve and impacts lifelong female reproduction, yet its developmental regulation remains elusive. Early progenitors or germline stem cells (GSCs) are not maintained in adult mammalian ovaries, and in mice they are lost closely following embryonic sex determination. Fundamental knowledge of vertebrate ovarian GSCs and how they are regulated to produce oocytes in the developmental context of the ovary is lacking. Here, we uncover a previously unrecognized ovarian developmental microenvironment, the ovarian module, in developing zebrafish and human ovaries. We show zebrafish ovarian modules comprise the morphogenetic unit of oocyte production, harboring GSCs at the young tip, which generate oocyte precursors and early differentiating oocytes in a spatially linear manner. We define module morphology, resolve its developmental dynamics, recapitulate its single-cell transcriptomic architecture, and identify Notch signaling as a regulator of GSCs. We map Notch pathway components in modules, identifying GSC as sender cells and their progeny as receiver cells, and functionally demonstrate that Notch activity is required to suppress the GSC fate and to promote differentiation. Ovarian modules persist in adult zebrafish ovaries, highlighting a similar framework in oocyte production during homeostasis. We further demonstrate that modules are conserved in human fetal ovaries, where POU5F1/OCT4+ germ cells and differentiating oocytes are similarly organized, and Notch components are expressed in equivalent cell types, supporting the module as a fundamental strategy in oocyte production. Our findings propose new developmental principles for oogenesis and establish new paradigms in female GSC biology and for women’s reproductive regenerative medicine. Biological sciences/Developmental biology/Germline development/Oogenesis Biological sciences/Stem cells/Stem-cell differentiation Biological sciences/Developmental biology/Stem-cell niche Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryVideo8.avi Supplementary Video 8. Supporting information for Fig. 3j. SupplementaryVideo2.mp4 Supplementary Video 2. Supporting information for Fig. 1h. SupplementaryVideo7.avi Supplementary Video 7. Supporting information for Fig. 3j. SupplementaryVideo6.avi Supplementary Video 6. Supporting information for Fig. 3j. SupplementaryVideo9.avi Supplementary Video 9. Supporting information for Fig. 3j. SupplementaryVideo4.mp4 Supplementary Video 4. Supporting information for Fig. 1l. SupplementaryVideo5.mp4 Supplementary Video 5. Supporting information for Fig. 1l. SupplementaryVideo3.mp4 Supplementary Video 3. Supporting information for Fig. 1k. SupplementaryVideo1.mp4 Supplementary Video 1. Supporting information for Fig. 1e. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7325126","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Biological Sciences - Article","associatedPublications":[],"authors":[{"id":534548136,"identity":"00493312-6d1a-4db3-bb48-a88513b64ba7","order_by":0,"name":"Yaniv 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Early progenitors or germline stem cells (GSCs) are not maintained in adult mammalian ovaries, and in mice they are lost closely following embryonic sex determination. Fundamental knowledge of vertebrate ovarian GSCs and how they are regulated to produce oocytes in the developmental context of the ovary is lacking. Here, we uncover a previously unrecognized ovarian developmental microenvironment, the ovarian module, in developing zebrafish and human ovaries. We show zebrafish ovarian modules comprise the morphogenetic unit of oocyte production, harboring GSCs at the young tip, which generate oocyte precursors and early differentiating oocytes in a spatially linear manner. We define module morphology, resolve its developmental dynamics, recapitulate its single-cell transcriptomic architecture, and identify Notch signaling as a regulator of GSCs. We map Notch pathway components in modules, identifying GSC as sender cells and their progeny as receiver cells, and functionally demonstrate that Notch activity is required to suppress the GSC fate and to promote differentiation. Ovarian modules persist in adult zebrafish ovaries, highlighting a similar framework in oocyte production during homeostasis. We further demonstrate that modules are conserved in human fetal ovaries, where POU5F1/OCT4+ germ cells and differentiating oocytes are similarly organized, and Notch components are expressed in equivalent cell types, supporting the module as a fundamental strategy in oocyte production. 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