Live imaging and conditional disruption of native vertebrate planar cell polarity

Live imaging and conditional disruption of native vertebrate planar cell polarity | 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 Live imaging and conditional disruption of native vertebrate planar cell polarity Maria Jussila, Curtis Boswell, Brian Ciruna This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-764931/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 23 Sep, 2022 Read the published version in Nature Communications → Version 1 posted You are reading this latest preprint version Abstract Tissue-wide coordination of polarized cytoskeletal organization and cell behaviour, critical for organ morphogenesis and tumour progression, is controlled by asymmetric membrane localization of non-canonical Wnt/planar cell polarity (PCP) signalling components. Understanding the dynamic regulation of PCP thus requires visualization of these core proteins. In vertebrates, immunohistochemical studies have provided snapshots of PCP within tissues while exogenous, fluorescently-tagged reporters have been introduced for live imaging of cell polarity. However, the functionality and spatiotemporal relevance of static and exogenous PCP readouts remain uncertain. Here we fluorescently tag an endogenous core PCP component, Vangl2, in zebrafish. We report on the authentic regulation of vertebrate PCP, through live imaging of this native sfGFP-Vangl2 protein during embryogenesis. We couple sfGFP-Vangl2 with conditional zGrad GFP-nanobody degradation methodologies for tissue-specific interrogation of PCP function. Together, our studies provide crucial insights into the establishment and maintenance of vertebrate PCP and create a powerful experimental paradigm for future investigations. General Cell Biology & Physiology Stem Cell & Developmental Cell Biology Full Text Additional Declarations There is NO Competing Interest. Supplementary Files JussilaetalMovie3NatComm.mp4 Supplementary Movie 3 JussilaetalMovie2NatComm.mp4 Supplementary Movie 2 JussilaetalSupplementaryFiguresNatComm.pdf JussilaetalMovie1NatComm.mp4 Supplementary Movie 1 JussilaetalMovie4NatComm.mp4 Supplementary Movie 4 JussilaetalSupplementaryFiguresNatComm.pdf Supplementary Figures 1-4 nrreportingsummaryNCOMMS2129897.pdf Reporting Summary Cite Share Download PDF Status: Published Journal Publication published 23 Sep, 2022 Read the published version in Nature Communications → 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. 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