Buffering of nuclear membrane tension and mechanotransduction by the endoplasmic reticulum revealed by quantitative ALPIN imaging

Buffering of nuclear membrane tension and mechanotransduction by the endoplasmic reticulum revealed by quantitative ALPIN imaging | 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 Letter Buffering of nuclear membrane tension and mechanotransduction by the endoplasmic reticulum revealed by quantitative ALPIN imaging Philipp Niethammer, Zhouyang Shen, Zaza Gelashvili This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5530637/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 09 Dec, 2025 Read the published version in Nature Cell Biology → Version 1 posted You are reading this latest preprint version Abstract Nuclear deformation by osmotic shock or necrosis activates the cytosolic phospholipase A2 (cPla2) nuclear shape sensing pathway, a key regulator of tissue inflammation and repair. Ca²⁺ and inner nuclear membrane (INM) tension (T INM ) are believed to mediate nucleoplasmic cPla2 activation. The concept implies that T INM persists long enough to stimulate cPla2-INM adsorption. However, T INM may instead be rapidly dissipated by the contiguous endoplasmic reticulum (ER), with cPla2-INM adsorption reporting rather on changes in Ca²⁺ than T INM . The impact of T INM and ER contiguity on nuclear shape sensing and mechanotransduction remains unknown. To address this gap, we developed the Ca 2+ insensitive, T INM -only biosensor ALPIN (Amphipathic Lipid Packing sensor domain Inside the Nucleus). By quantitative ALPIN imaging, we found that stress-induced ER fragmentation increases T INM and nuclear membrane mechanotransduction in osmotically shocked or ferroptotic cells, permeabilized cell corpses, and at zebrafish wounds in vivo. Our findings reveal critical roles for the ER and T INM in nuclear shape sensing and introduce ALPIN as promising tool for studying organelle membrane mechanotransduction in health and disease. Biological sciences/Cell biology/Organelles/Nucleus Biological sciences/Biological techniques/Imaging/Fluorescence imaging cPLA2 ALPS nuclear membrane tension mechanotransduction endoplasmic reticulum ferroptosis wound Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryVideo1.avi Supplementary Video 1 SupplementaryVideo2.avi Supplementary Video 2 SupplementaryVideo3compressed.avi Supplementary Video 3 Cite Share Download PDF Status: Published Journal Publication published 09 Dec, 2025 Read the published version in Nature Cell Biology → 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-5530637","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Letter","associatedPublications":[],"authors":[{"id":387271965,"identity":"4aa7fc4c-7553-4c61-ade7-a79754372a5a","order_by":0,"name":"Philipp 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endoplasmic reticulum revealed by quantitative ALPIN imaging","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"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":"cPLA2, ALPS, nuclear membrane tension, mechanotransduction, endoplasmic reticulum, ferroptosis, wound","lastPublishedDoi":"10.21203/rs.3.rs-5530637/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5530637/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Nuclear deformation by osmotic shock or necrosis activates the cytosolic phospholipase A2 (cPla2) nuclear shape sensing pathway, a key regulator of tissue inflammation and repair. Ca²⁺ and inner nuclear membrane (INM) tension (T\u003csub\u003eINM\u003c/sub\u003e) are believed to mediate nucleoplasmic cPla2 activation. The concept implies that T\u003csub\u003eINM\u003c/sub\u003e persists long enough to stimulate cPla2-INM adsorption. However, T\u003csub\u003eINM\u003c/sub\u003e may instead be rapidly dissipated by the contiguous endoplasmic reticulum (ER), with cPla2-INM adsorption reporting rather on changes in Ca²⁺ than T\u003csub\u003eINM\u003c/sub\u003e. The impact of T\u003csub\u003eINM\u003c/sub\u003e and ER contiguity on nuclear shape sensing and mechanotransduction remains unknown.\r\nTo address this gap, we developed the Ca\u003csup\u003e2+\u003c/sup\u003e insensitive, T\u003csub\u003eINM\u003c/sub\u003e-only biosensor ALPIN (Amphipathic Lipid Packing sensor domain Inside the Nucleus). By quantitative ALPIN imaging, we found that stress-induced ER fragmentation increases T\u003csub\u003eINM\u003c/sub\u003e and nuclear membrane mechanotransduction in osmotically shocked or ferroptotic cells, permeabilized cell corpses, and at zebrafish wounds in vivo. Our findings reveal critical roles for the ER and T\u003csub\u003eINM\u003c/sub\u003e in nuclear shape sensing and introduce ALPIN as promising tool for studying organelle membrane mechanotransduction in health and disease.","manuscriptTitle":"Buffering of nuclear membrane tension and mechanotransduction by the endoplasmic reticulum revealed by quantitative ALPIN imaging","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-12-09 02:52:07","doi":"10.21203/rs.3.rs-5530637/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-cell-biology","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"ncb","sideBox":"Learn more about [Nature Cell Biology](http://www.nature.com/ncb/)","snPcode":"","submissionUrl":"","title":"Nature Cell Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Research","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"26fec094-6953-4060-992b-6bcd05839681","owner":[],"postedDate":"December 9th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":41274276,"name":"Biological sciences/Cell biology/Organelles/Nucleus"},{"id":41274277,"name":"Biological sciences/Biological techniques/Imaging/Fluorescence imaging"}],"tags":[],"updatedAt":"2025-12-10T08:09:19+00:00","versionOfRecord":{"articleIdentity":"rs-5530637","link":"https://doi.org/10.1038/s41556-025-01820-9","journal":{"identity":"nature-cell-biology","isVorOnly":false,"title":"Nature Cell Biology"},"publishedOn":"2025-12-09 05:00:00","publishedOnDateReadable":"December 9th, 2025"},"versionCreatedAt":"2024-12-09 02:52:07","video":"","vorDoi":"10.1038/s41556-025-01820-9","vorDoiUrl":"https://doi.org/10.1038/s41556-025-01820-9","workflowStages":[]},"version":"v1","identity":"rs-5530637","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5530637","identity":"rs-5530637","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}