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Live imaging and conditional disruption of native PCP activity using endogenously tagged zebrafish sfGFP-Vangl2

Author

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  • Maria Jussila

    (The Hospital for Sick Children)

  • Curtis W. Boswell

    (The Hospital for Sick Children
    The University of Toronto
    Yale University School of Medicine)

  • Nigel W. Griffiths

    (The Hospital for Sick Children)

  • Patrick G. Pumputis

    (The Hospital for Sick Children
    The University of Toronto)

  • Brian Ciruna

    (The Hospital for Sick Children
    The University of Toronto)

Abstract

Tissue-wide coordination of polarized cytoskeletal organization and cell behaviour, critical for normal development, 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 polarity proteins in vivo. Here we utilize CRISPR/Cas9 genome editing to introduce a fluorescent reporter onto the core PCP component, Vangl2, in zebrafish. Through live imaging of endogenous sfGFP-Vangl2 expression, we report on the authentic regulation of vertebrate PCP during embryogenesis. Furthermore, we couple sfGFP-Vangl2 with conditional zGrad GFP-nanobody degradation methodologies to interrogate tissue-specific functions for PCP. Remarkably, loss of Vangl2 in foxj1a-positive cell lineages causes ependymal cell cilia and Reissner fiber formation defects as well as idiopathic-like scoliosis. Together, our studies provide crucial insights into the establishment and maintenance of vertebrate PCP and create a powerful experimental paradigm for investigating post-embryonic and tissue-specific functions for Vangl2 in development and disease.

Suggested Citation

  • Maria Jussila & Curtis W. Boswell & Nigel W. Griffiths & Patrick G. Pumputis & Brian Ciruna, 2022. "Live imaging and conditional disruption of native PCP activity using endogenously tagged zebrafish sfGFP-Vangl2," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-33322-9
    DOI: 10.1038/s41467-022-33322-9
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    References listed on IDEAS

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    1. Carl-Philipp Heisenberg & Masazumi Tada & Gerd-Jörg Rauch & Leonor Saúde & Miguel L. Concha & Robert Geisler & Derek L. Stemple & James C. Smith & Stephen W. Wilson, 2000. "Silberblick/Wnt11 mediates convergent extension movements during zebrafish gastrulation," Nature, Nature, vol. 405(6782), pages 76-81, May.
    2. Jose Maria Carvajal-Gonzalez & Angel-Carlos Roman & Marek Mlodzik, 2016. "Positioning of centrioles is a conserved readout of Frizzled planar cell polarity signalling," Nature Communications, Nature, vol. 7(1), pages 1-9, September.
    3. Brian Ciruna & Andreas Jenny & Diana Lee & Marek Mlodzik & Alexander F. Schier, 2006. "Planar cell polarity signalling couples cell division and morphogenesis during neurulation," Nature, Nature, vol. 439(7073), pages 220-224, January.
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