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

Maria Jussila, Curtis W. Boswell, Nigel W. Griffiths, Patrick G. Pumputis and Brian Ciruna ()
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Maria Jussila: The Hospital for Sick Children
Curtis W. Boswell: The Hospital for Sick Children
Nigel W. Griffiths: The Hospital for Sick Children
Patrick G. Pumputis: The Hospital for Sick Children
Brian Ciruna: The Hospital for Sick Children

Nature Communications, 2022, vol. 13, issue 1, 1-13

Abstract: 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.

Date: 2022
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DOI: 10.1038/s41467-022-33322-9

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