Wnt Signaling pp 207-219 | Cite as

Detection of Planar Polarity Proteins in Mammalian Cochlea

  • Mireille Montcouquiol
  • Jennifer M. Jones
  • Nathalie Sans
Part of the Methods in Molecular Biology™ book series (MIMB, volume 468)

Abstract

The “core genes” were identified as a group of genes believed to function as a conserved signaling cassette for the specification of planar polarity in Drosophila Melanogaster, and includes frizzled (fz), van gogh (vang) or strabismus (stbm), prickle (Pk), dishevelled (dsh), flamingo (fmi), and diego. The mutation of each of these genes not only causes the disruption of planar polarity within the wing or the eye of the animal, but also affects the localization of all the other protein members of the core group. These properties emphasize the importance of the interrelations between the proteins of this group. All of these core genes have homologs in vertebrates. Studies in Danio Rerio (zebrafish) and Xenopus laevis (frog) have uncovered other roles for some of these molecules in gastrulation and neurulation, during which the shape of a given tissue will undergo major transformation through cell movements. A disruption in these processes can lead to severe neural tube defects in diverse organisms, including humans. In fact, a large body of evidence suggests that planar polarity proteins are not involved in one specific cascade but in many different ones and many different mechanisms such as, but not limited to, hair or cilia orientation, asymmetric division, cellular movements, or neuronal migration.

In mice cochleae, mutations in planar polarity genes lead to defects in the orientation of the stere-ociliary bundles at the apex of each hair cell. This phenotype established the cochlea as one of the clearest examples of planar polarity in mammals. Although significant progress has been made toward understanding the molecular basis required for the development of planar polarity in invertebrates, similar advances in vertebrates are more recent and rely mainly on the identification of a group of mammalian mutants that affect hair cell stereociliary bundle orientation. These include mutation of vangl2, scrb1, celsr1, PTK-7, dvl1-2, and more recently fz3 and fz6 (1). In this chapter, we describe how to use the mammalian cochlea, which represents one of the best systems to study planar polarity in mammals, to identify planar polarity mutants, study protein distribution, do in vitro analysis, and perform Western blots to analyze putative planar polarity proteins.

Key words

Vangl2 Planar polarity Asymmetry Culture Cochlea Immunofluorescence 

Notes

Acknowledgments

We thank F. Loll for technical assistance, Elodie Richard for proofreading, and Dr. Ronna Hertzano for the photographs in Fig. 16.1. This work was supported by grants from Institut National de la Santé et de la Recherche Médicale, Fondation pour la Recherche Médicale, and Région Aquitaine (MM and NS).

References

  1. 1.
    Montcouquiol, M., Crenshaw, E. B., and Kelley, M.W. (2006) Non canonical Wnt signalling and neural polarity. Annu Rev Neurosci. 29, 363–386.CrossRefPubMedGoogle Scholar
  2. 2.
    Eaton, S. (1997) Planar polarization of Dro-sophila and vertebrate epithelia. Curr. Opin. Cell Biol. 9, 860–866.CrossRefPubMedGoogle Scholar
  3. 3.
    Montcouquiol, M., Rachel, R. A., Lan-ford P. J., Copeland, N. G., Jenkins, N, A., and Kelley, M.W. (2003) Identification of Vangl2 and Scrb1 as planar polarity genes in mammals. Nature 423, 173–177.CrossRefPubMedGoogle Scholar
  4. 4.
    Wang, Y., Guo, N., and Nathans, J. (2006) The role of Frizzled3 and Frizzled6 in neural tube closure and in the planar polarity of inner-ear sensory hair cells. J. Neurosci. 26, 2147–2156.CrossRefPubMedGoogle Scholar
  5. 5.
    Montcouquiol, M., Sans, N., Huss, D., Kach, J., Dickman, J.D., Forge, A., Rachel, R.A., Copeland, N. G., Jenkins, N. A., Bogani, D., Murdoch, J., Warchol, M. E., Wenthold, R.J., and Kelley, M. W. (2006) Asymmetric localization of Vangl2 and Fz3 indicate novel mechanisms for planar cell polarity in mammals. J. Neurosci. 26, 5265–5275.CrossRefPubMedGoogle Scholar
  6. 6.
    Kaufman, M. H. (1992) The atlas of mouse development. Academic Press, San Diego, California. 386 pp.Google Scholar
  7. 7.
    Jones, J. M., Montcouquiol, M., Dab-doub, A., Woods, C., and Kelley, M. W. (2006) Inhibitors of differentiation and DNA binding (Ids) regulate Math1 and hair cell formation during the development of the organ of Corti. J. Neurosci. 26, 550–558.CrossRefPubMedGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Mireille Montcouquiol
    • 1
  • Jennifer M. Jones
    • 2
  • Nathalie Sans
    • 3
  1. 1.Equipe Avenir, Development Neurosciences, INSERM U862Institut Francois Magendie, Université Bordeaux IIBordeaux CédexFrance
  2. 2.Department of OtolaryngologyWashington University School of MedicineUSA
  3. 3.Equipe Avenir, Molecular Neurobiology, INSERM U862Université Bordeaux IIBordeaux CedexFrance

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