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Analysis of PCP Defects in Mammalian Eye Lens

  • Yuki Sugiyama
  • John W. McAvoyEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 839)

Abstract

Multicellular tissues and organs often show planar cell polarity (PCP) where the constituent cells align along an axis to form coordinated patterns. Mammalian eye lenses are mainly comprised of epithelial-derived fibre cells, which exhibit highly ordered alignment that is regulated by PCP signaling. Each fibre cell has an apically situated primary cilium and in most cases this is polarized towards the lens anterior pole. Here we describe how to visualize the global cellular alignment of lens fibre cells by examining the suture pattern that is formed by the tips of fibres meeting at the anterior pole. We also describe a method for whole mount preparation, which allows observation of the polarized distribution of primary cilia at the apical surface of lens fibres. Given its relative simplicity, at least in cellular terms, and its requirement for a high degree of precision in cellular alignment and orientation, we predict that the lens will be an excellent model system to help elucidate the role of cilia and PCP components in the development of three-dimensional organization in tissues and organs.

Key words

Planar cell polarity (PCP) Mammalian eye lens Lens fibre cell Lens suture Primary cilium Whole mount 

Notes

Acknowledgements

This work was supported by NHMRC (Australia), NIH (USA, R01 EY03177), ORIA, Australia and The Sydney Foundation for Medical Research. Y.S. was supported by an Endeavour Fellowship, Australia and The Sydney Eye Hospital Foundation. Some research illustrated here was undertaken as part of the Vision CRC, New South Wales, Sydney, Australia.

References

  1. 1.
    Lawrence PA, Struhl G, Casal J (2007) Planar cell polarity: one or two pathways. Nat Rev Genet 8:555–563.PubMedCrossRefGoogle Scholar
  2. 2.
    Seifert JR, Mlodzik M (2007) Frizzled/PCP signaling: a conserved mechanism regulating cell polarity and directed motility. Nat Rev Genet 8:126–138.PubMedCrossRefGoogle Scholar
  3. 3.
    Strutt D (2008) The planar polarity pathway. Curr Biol 18:R898–902.PubMedCrossRefGoogle Scholar
  4. 4.
    Wang Y, Nathans J (2007) Tissue/planar cell polarity in vertebrates: new insights and new questions. Development 134:647–658.PubMedCrossRefGoogle Scholar
  5. 5.
    Axelrod JD (2008) Basal bodies, kinocilia and planar cell polarity. Nat Genet 40:10–11.PubMedCrossRefGoogle Scholar
  6. 6.
    Park TJ, Haigo SL, Wallingford JB (2006) Ciliogenesis defects in embryos lacking inturned or fuzzy function are associated with failure of planar cell polarity and Hedgehog signaling. Nat Genet 38:303–311.PubMedCrossRefGoogle Scholar
  7. 7.
    Ross AJ, May-Simera H, Eichers ER, Kai M, Hill J, Jagger DJ, Leitch CC, Chapple JP, Munro PM, Fisher S, Tan PL, Phillips HM, Leroux MR, Henderson DJ, Murdoch JN, Copp AJ, Eliot MM, Lupski JR, Kemp DT, Dolfus H, Tada M, Katsanis N, Forge A, Beales PL (2005) Disruption of Bardet–Biedl syndrome ciliary proteins perturbs planar cell polarity in vertebrates. Nat Genet 37:1135–1140.PubMedCrossRefGoogle Scholar
  8. 8.
    Simons M, Walz G (2006) Polycystic kidney disease: cell division without a c(l)ue? Kidney Int 70:854–864.PubMedCrossRefGoogle Scholar
  9. 9.
    Singla V, Reiter JF (2006) The primary cilium as the cell’s antenna: signaling at a sensory organelle. Science 313:629–633.PubMedCrossRefGoogle Scholar
  10. 10.
    Sugiyama Y, Stump RJ, Nguyen A, Wen L, Chen Y, Wang Y, Murdoch JN, Lovicu FJ, McAvoy JW (2010) Secreted frizzled-related protein disrupts PCP in eye lens fiber cells that have polarized primary cilia. Dev Biol 338, 193–201.PubMedCrossRefGoogle Scholar
  11. 11.
    Chen Y, Stump RJ, Lovicu FJ, Shimono A, McAvoy JW (2008) Wnt signaling is required for organization of the lens fiber cell cytoskeleton and development of lens three-dimensional architecture. Dev Biol 324:161–176.PubMedCrossRefGoogle Scholar
  12. 12.
    Ashery-Padan R, Marquardt T, Zhou, X, Gruss P (2000) Pax6 activity in the lens primordium is required for lens formation and for correct placement of a single retina in the eye. Genes Dev 14:2701–2711.PubMedCrossRefGoogle Scholar
  13. 13.
    Zhao H, Yang Y, Rizo CM, Overbeek PA, Robinson ML (2004) Insertion of a Pax6 consensus binding site into the alphaA-crystallin promoter acts as a lens epithelial cell enhancer in transgenic mice. Invest Ophthalmol Vis Sci 45:1930–1939.PubMedCrossRefGoogle Scholar
  14. 14.
    Lovicu FJ, McAvoy JW (2008) Epithelial explants and their application to study developmental processes in the lens. In: Tsonis PA (ed) Animal Models in Eye Research, Academic Press, New York.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Save Sight InstituteThe University of SydneySydneyAustralia
  2. 2.Sydney Hospital and Eye HospitalSydneyAustralia

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