Linking Centromere Stretch to SAC Signaling in Mammalian Oocytes: Insights from a Kinetochore-Modified Model

Linking Centromere Stretch to SAC Signaling in Mammalian Oocytes: Insights from a Kinetochore-Modified Model | 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 Article Linking Centromere Stretch to SAC Signaling in Mammalian Oocytes: Insights from a Kinetochore-Modified Model Yan Yun, Simon Lane, Ziyang Guo, Zhixiang Zheng, Janet Holt, Keith Jones This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8643946/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract The spindle assembly checkpoint (SAC) monitors kinetochore-microtubule interactions to ensure accurate chromosome segregation. In mitosis, investigation of its silencing at anaphase centers on whether attachment or tension is the key signal, while in meiosis for mammalian oocytes it remains unclear whether centromere stretching contributes to SAC deactivation, because in this reductional division sister chromatids co-segregate during meiosis I. While previous studies have linked bivalent tension to SAC control in oocytes, this measurement cannot be directly equated with mitotic chromatin stretching. Here, by tagging the N-terminus of NDC80/HEC1, we developed a mouse oocyte model that features stable microtubule attachments, yet exhibits persistent SAC signaling at metaphase I. Our comprehensive analysis revealed a strong correlation between centromere stretching and SAC signaling. Furthermore, high-resolution live imaging demonstrated that these modified kinetochores impaired their oscillatory movements, a process vital for error correction, while chromosome congression remained largely unaffected. Collectively, our data suggest that stable microtubule attachment alone is insufficient to silence SAC in mammalian oocytes; instead, centromere stretching may serve as a critical signal to deactivate the checkpoint. These findings underscore the adaptability of core cell-cycle surveillance pathways. Biological sciences/Cell biology/Cell division/Checkpoints Biological sciences/Cell biology/Chromosomes/Kinetochores Biological sciences/Cell biology/Cell division/Meiosis Biological sciences/Cell biology/Cellular imaging/Super-resolution microscopy Microtubule-kinetochore interaction centromere stretch spindle assembly checkpoint NDC80/HEC1 oocytes Full Text Additional Declarations There is NO Competing Interest. Supplementary Files MovieS1ControlKT.avi Oocytes with control kinetochores underwent bivalent segregation at anaphase I MovieS2ModifiedKT.avi Oocytes with modified kinetochores failed to complete meiosis I MovieS3convsn80n.avi The kinetochore modification inhibited oscillatory movement of sister kinetochore pairs in mouse oocytes Cite Share Download PDF Status: Under Review 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-8643946","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":585531658,"identity":"dce476c8-bad5-4e37-b2d4-96d47b4a2dc5","order_by":0,"name":"Yan Yun","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA00lEQVRIiWNgGAWjYPACGyBOACMGhgPEaUkjXcthBqh6IrQYHD97TJp3x/nE/vbkZxIP/jDI8d1IIKDlTF6aNO+Z24kzzjwzk0hsYzCWJKjlQI6ZNG/b7cQNEgnGBokNDIkbCGo5/wak5RxQS/png4Q/DPWEtdwA23IAqCXH8EECG0OCASEtkjfeGFvObUs2nnHmTeGDxDYJw5lnHuDXwnc+x/DG2zY72f729A0Hf/yxkec7TsAWhQMMLBJIfAmcKuFAvoGB+QNhZaNgFIyCUTCiAQAJR0tala8rTgAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-8293-0070","institution":"Shantou Central Hospital","correspondingAuthor":true,"prefix":"","firstName":"Yan","middleName":"","lastName":"Yun","suffix":""},{"id":585531659,"identity":"7282c8ae-fc33-476e-b817-1ebddf5f03de","order_by":1,"name":"Simon Lane","email":"","orcid":"https://orcid.org/0000-0002-8155-0981","institution":"University of Southampton","correspondingAuthor":false,"prefix":"","firstName":"Simon","middleName":"","lastName":"Lane","suffix":""},{"id":585531660,"identity":"84bb8613-157f-4ec4-858c-66f080d0b734","order_by":2,"name":"Ziyang Guo","email":"","orcid":"","institution":"Shantou University Medical College","correspondingAuthor":false,"prefix":"","firstName":"Ziyang","middleName":"","lastName":"Guo","suffix":""},{"id":585531661,"identity":"c4a0ff9b-0dd1-4d17-9ff3-7abaca074f86","order_by":3,"name":"Zhixiang Zheng","email":"","orcid":"","institution":"Shantou Central Hospital","correspondingAuthor":false,"prefix":"","firstName":"Zhixiang","middleName":"","lastName":"Zheng","suffix":""},{"id":585531662,"identity":"95023f48-6579-402c-8da7-e6e29d19d875","order_by":4,"name":"Janet Holt","email":"","orcid":"","institution":"University of Newcastle","correspondingAuthor":false,"prefix":"","firstName":"Janet","middleName":"","lastName":"Holt","suffix":""},{"id":585531663,"identity":"0b33f555-eea5-4e10-9b07-e82c04b36058","order_by":5,"name":"Keith Jones","email":"","orcid":"","institution":"University of Sussex","correspondingAuthor":false,"prefix":"","firstName":"Keith","middleName":"","lastName":"Jones","suffix":""}],"badges":[],"createdAt":"2026-01-20 02:16:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8643946/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8643946/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":105568944,"identity":"5e8cbef5-e252-4c2a-a525-38454feb6e08","added_by":"auto","created_at":"2026-03-27 13:10:54","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2731337,"visible":true,"origin":"","legend":"Article File","description":"","filename":"ManuscriptYun.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8643946/v1_covered_0d827caa-f6e0-4cc5-90c6-093f3e6594e3.pdf"},{"id":105562979,"identity":"334182d7-84ec-422d-8a78-7e6eb23b0c47","added_by":"auto","created_at":"2026-03-27 12:45:28","extension":"avi","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":400644,"visible":true,"origin":"","legend":"Oocytes with control kinetochores underwent bivalent segregation at anaphase I","description":"","filename":"MovieS1ControlKT.avi","url":"https://assets-eu.researchsquare.com/files/rs-8643946/v1/1b3fb9f333c986f8542d712d.avi"},{"id":105091977,"identity":"608b8daf-e32f-43cb-9e8c-c283754e9146","added_by":"auto","created_at":"2026-03-21 00:35:45","extension":"avi","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":755474,"visible":true,"origin":"","legend":"Oocytes with modified kinetochores failed to complete meiosis I","description":"","filename":"MovieS2ModifiedKT.avi","url":"https://assets-eu.researchsquare.com/files/rs-8643946/v1/e8107e565d3713b4ca7c56d6.avi"},{"id":105563132,"identity":"6c7586f2-5c7b-4439-a0a0-dfd38fd1a6f0","added_by":"auto","created_at":"2026-03-27 12:46:04","extension":"avi","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":1309412,"visible":true,"origin":"","legend":"The kinetochore modification inhibited oscillatory movement of sister kinetochore pairs in mouse oocytes","description":"","filename":"MovieS3convsn80n.avi","url":"https://assets-eu.researchsquare.com/files/rs-8643946/v1/024af853f23e60fbcb3e5afc.avi"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Linking Centromere Stretch to SAC Signaling in Mammalian Oocytes: Insights from a Kinetochore-Modified Model","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Microtubule-kinetochore interaction, centromere stretch, spindle assembly checkpoint, NDC80/HEC1, oocytes","lastPublishedDoi":"10.21203/rs.3.rs-8643946/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8643946/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The spindle assembly checkpoint (SAC) monitors kinetochore-microtubule interactions to ensure accurate chromosome segregation. In mitosis, investigation of its silencing at anaphase centers on whether attachment or tension is the key signal, while in meiosis for mammalian oocytes it remains unclear whether centromere stretching contributes to SAC deactivation, because in this reductional division sister chromatids co-segregate during meiosis I. While previous studies have linked bivalent tension to SAC control in oocytes, this measurement cannot be directly equated with mitotic chromatin stretching. Here, by tagging the N-terminus of NDC80/HEC1, we developed a mouse oocyte model that features stable microtubule attachments, yet exhibits persistent SAC signaling at metaphase I. Our comprehensive analysis revealed a strong correlation between centromere stretching and SAC signaling. Furthermore, high-resolution live imaging demonstrated that these modified kinetochores impaired their oscillatory movements, a process vital for error correction, while chromosome congression remained largely unaffected. Collectively, our data suggest that stable microtubule attachment alone is insufficient to silence SAC in mammalian oocytes; instead, centromere stretching may serve as a critical signal to deactivate the checkpoint. These findings underscore the adaptability of core cell-cycle surveillance pathways.","manuscriptTitle":"Linking Centromere Stretch to SAC Signaling in Mammalian Oocytes: Insights from a Kinetochore-Modified Model","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-21 00:35:40","doi":"10.21203/rs.3.rs-8643946/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"communications-biology","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"commsbio","sideBox":"Learn more about [Communications Biology](http://www.nature.com/commsbio/)","snPcode":"","submissionUrl":"","title":"Communications Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Communications Series","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"026b3794-9cb6-451b-b399-3735c4eb45d1","owner":[],"postedDate":"March 21st, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":62293087,"name":"Biological sciences/Cell biology/Cell division/Checkpoints"},{"id":62293088,"name":"Biological sciences/Cell biology/Chromosomes/Kinetochores"},{"id":62293089,"name":"Biological sciences/Cell biology/Cell division/Meiosis"},{"id":62293090,"name":"Biological sciences/Cell biology/Cellular imaging/Super-resolution microscopy"}],"tags":[],"updatedAt":"2026-03-21T00:35:40+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-21 00:35:40","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8643946","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8643946","identity":"rs-8643946","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}