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Vangl2 directs the posterior tilting and asymmetric localization of motile primary cilia

Nature Cell Biology volume 12, pages 407–412 (2010)Cite this article

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Abstract

Cilia are microtubule-based organelles that project into the extracellular space, function in the perception and integration of environmental cues1, and regulate Hedgehog signal transduction2. The emergent association of ciliary defects with diverse and pleiotropic human disorders3,4 has fuelled investigations into the molecular genetic regulation of ciliogenesis. Although recent studies implicate planar cell polarity (PCP) in cilia formation, this conclusion is based on analyses of proteins that are not specific to, or downstream effectors of PCP signal transduction5,6,7,8,9,10. Here we characterize zebrafish embryos devoid of all Vangl2 function11, a core and specific component of the PCP signalling pathway. Using Arl13b–GFP as a live marker of the ciliary axoneme, we demonstrate that Vangl2 is not required for ciliogenesis. Instead, Vangl2 controls the posterior tilting of primary motile cilia lining the neurocoel, Kupffer's vesicle and pronephric duct. Furthermore, we show that Vangl2 is required for asymmetric localization of cilia to the posterior apical membrane of neuroepithelial cells. Our results indicate a broad and essential role for PCP in the asymmetric localization and orientation of motile primary cilia, establishing directional fluid flow implicated in normal embryonic development and disease.

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Figure 1: MZvangl2 mutants show left–right patterning defects, irregular fluid flow and abnormal cilia orientation within Kupffer's vesicle (KV).
Figure 2: Arl13b–GFP expression reveals asymmetric orientation and position of motile primary cilia at the floorplate.
Figure 3: Chimaeric analysis reveals a cell autonomous function for Vangl2 in controlling the posterior tilting of motile floorplate cilia.
Figure 4: MZvangl2 mutants show normal differentiation of multiciliated cells in the pronephric duct, but show defects in the orientation of primary motile cilia.
Figure 5: Analysis of MZknypek mutant embryos reveal similar defects in the asymmetric localization and orientation of primary motile cilia.

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Acknowledgements

We thank Tamara Caspary for her gift of the mArl13b cDNA clone; Adrian Salic and Lila Solnica-Krezel for providing the Centrin–GFP expression construct; Sasha Fernando and Angela Morley for zebrafish husbandry; and Danielle Gelinas for expert technical support. This research was undertaken, in part, thanks to funding from the Terry Fox Foundation, the Natural Sciences and Engineering Research Council of Canada, and the Canada Research Chairs program.

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  1. Daniel Voskas

    Present address: Current address: Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada, M5G 1X5.,

Authors and Affiliations

  1. Program in Developmental & Stem Cell Biology, The Hospital for Sick Children, Toronto, M5G 1X8, Ontario, Canada

    Antonia Borovina, Simone Superina, Daniel Voskas & Brian Ciruna

  2. Department of Molecular Genetics, University of Toronto, Toronto, M5S 1A8, Ontario, Canada

    Antonia Borovina, Simone Superina & Brian Ciruna

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  1. Antonia Borovina
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Contributions

S.S. generated MZkny mutants; S.S. and B.C. characterized KV cilia orientation; D.V. and S.S. quantified left–right patterning defects; A.B. and D.V. characterized Arl13b localization in wild-type and MZvangl2 mutants. A.B. performed all further experimentation and analysis; B.C. supervised the project and wrote the manuscript.

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Correspondence to Brian Ciruna.

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Borovina, A., Superina, S., Voskas, D. et al. Vangl2 directs the posterior tilting and asymmetric localization of motile primary cilia. Nat Cell Biol 12, 407–412 (2010). https://doi.org/10.1038/ncb2042

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