Letter | Published:

Primary cilium migration depends on G-protein signalling control of subapical cytoskeleton

Nature Cell Biology volume 15, pages 11071115 (2013) | Download Citation

Abstract

In ciliated mammalian cells, the precise migration of the primary cilium at the apical surface of the cells, also referred to as translational polarity, defines planar cell polarity (PCP) in very early stages. Recent research has revealed a co-dependence between planar polarization of some cell types and cilium positioning at the surface of cells. This important role of the primary cilium in mammalian cells is in contrast with its absence from Drosophila melanogaster PCP establishment. Here, we show that deletion of GTP-binding protein alpha-i subunit 3 (Gαi3) and mammalian Partner of inscuteable (mPins) disrupts the migration of the kinocilium at the surface of cochlear hair cells and affects hair bundle orientation and shape. Inhibition of G-protein function in vitro leads to kinocilium migration defects, PCP phenotype and abnormal hair bundle morphology. We show that Gαi3/mPins are expressed in an apical and distal asymmetrical domain, which is opposite and complementary to an aPKC/Par-3/Par-6b expression domain, and non-overlapping with the core PCP protein Vangl2. Thus G-protein-dependent signalling controls the migration of the cilium cell autonomously, whereas core PCP signalling controls long-range tissue PCP.

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Acknowledgements

We thank the animal and genotyping facilities’ members of the Neurocentre for technical assistance, notably H. Doat and D. Gonzales. We also thank the entire team of the Bordeaux Imaging Center (BIC) for the constant technical assistance, notably P. Legros, S. Marais and C. Poujol. We thank P. Beales (UCL, UK) for the Mkks mutants. We thank L. Mays (Tubingen, Germany) for critical reading of the manuscript, and F. Schweisguth (Paris, France) and J. Raff (Oxford, UK) for thoughtful discussions. We apologize to all whose relevant work could not be cited.

This research was supported by an INSERM grant to M.M. and N.S., the Conseil Regional d’Aquitaine Neurocampus program, La Fondation pour la Recherche Medicale (M.M., N.S., J.E., A-C.L.), ANR-08-MNPS-040-01 (M.M.), the European Commission Coordination Action ENINET (LSHM-CT-2005-19063; N.S. and M.M.), Ligue Nationale Contre le Cancer (Label 2010, J-P.B.), EUCAAD (FP7 program, J-P.B.), Fondation ARC pour la Recherche sur le Cancer (B.N., E.B.), the Deutsche Forschungsgemeinschaft (DFG; B.N., S.B-H.), the Intramural Research Program of the NIH (Project Z01-ES-101643 to L.B.), the European FP7 program (HEALTH-F2-2008-200234, A.L.B.) and ANR (BLAN07-2-186738, A.L.B.).

Author information

Affiliations

  1. INSERM, Planar Polarity and Plasticity Group, Neurocentre Magendie, 33077 Bordeaux, France

    • Jerome Ezan
    • , Léa Lasvaux
    • , Aysegul Gezer
    • , Nathalie Sans
    •  & Mireille Montcouquiol
  2. Université Bordeaux, Neurocentre Magendie, Bordeaux 33077, France

    • Jerome Ezan
    • , Léa Lasvaux
    • , Aysegul Gezer
    • , Nathalie Sans
    •  & Mireille Montcouquiol
  3. Department of Pharmacology and Experimental Therapy, Institute of Experimental and Clinical Pharmacology and Toxicology, Eberhard Karls University Hospitals and Clinics, and Interfaculty Center of Pharmacogenomics and Drug Research, University of Tübingen, 72074 Tübingen, Germany

    • Ana Novakovic
    • , Sandra Beer-Hammer
    •  & Bernd Nürnberg
  4. Neurobiology-Neurodegeneration & Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892, USA

    • Helen May-Simera
  5. INSERM U1068, CRCM, 13009 Marseille, France

    • Edwige Belotti
    • , Anne-Catherine Lhoumeau
    •  & Jean-Paul Borg
  6. CNRS UMR7258, CRCM, 13009 Marseille, France

    • Edwige Belotti
    • , Anne-Catherine Lhoumeau
    •  & Jean-Paul Borg
  7. Institut Paoli-Calmettes, 13009 Marseille, France

    • Edwige Belotti
    • , Anne-Catherine Lhoumeau
    •  & Jean-Paul Borg
  8. Aix-Marseille Université, 13007 Marseille, France

    • Edwige Belotti
    • , Anne-Catherine Lhoumeau
    •  & Jean-Paul Borg
  9. Laboratory of Neurobiology, National Institute of Environmental Health Sciences, National Institutes of Health/Department of Health and Human Services, Durham, North Carolina 27709, USA

    • Lutz Birnbaumer
  10. CNRS, UMR 7288, Developmental Biology Institute of Marseille Luminy (IBDML), case 907, 13288 Marseille, cedex 09, France

    • André Le Bivic
  11. Aix-Marseille Université, Developmental Biology Institute of Marseille Luminy (IBDML), 13288 Marseille, France

    • André Le Bivic

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Contributions

M.M., J.E. and N.S. designed and carried out experiments, analysed data and wrote the paper. A.G. and L.L. carried out immunocytochemistry on cochleae, cultures and western blots. L.B., B.N., S.B-H., A.N. and A.L.B. generated and provided the G-protein mutant and G-protein antibodies, and carried out the characterization of the Go-protein antibody. E.B., A-C.L. and J-P.B. generated the rat anti-Vangl2 monoclonal antibody and PTK7-deficient mice. J-P.B., S.B-H., B.N. and A.L.B. provided experimental and conceptual advice and edited the manuscript. All authors discussed the results and implications and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Mireille Montcouquiol.

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DOI

https://doi.org/10.1038/ncb2819

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