Advertisement

Cuticle Refraction Microscopy: A Rapid and Simple Method for Imaging Drosophila Wing Topography, an Alternative Readout of Wing Planar Cell Polarity

  • David Neff
  • Justin Hogan
  • Simon CollierEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 839)

Abstract

The polarity of hairs on the adult Drosophila wing provides information about the planar cell polarity (PCP) signaling events that occur during pupal wing development. We have recently shown that PCP signaling also determines the orientation of cuticle ridges that traverse the surface of the adult wing membrane; a feature we call the wing membrane topography. Although hair polarity is uniform across the wild-type wing, ridge orientation differs between the anterior and posterior wing. Consequently, mapping wing topography can provide additional information about PCP signaling, rather than simply confirming observations of wing hair polarity. Wing membrane ridges can be imaged using scanning electron microscopy, however, significant preparation time and operator expertise are required. Here, we describe cuticle refraction microscopy, a rapid and simple light microscopy method for imaging Drosophila wing topography.

Key words

Drosophila Planar cell polarity Wing Hair Membrane Ridge Topography 

Notes

Acknowledgments

Thanks to Kristy Doyle for her contribution to the early development of the CRM method. Thanks also to Michael Norton at the Marshall University Molecular and Biological Imaging Center (MBIC) for assistance with SEM imaging. Development of the CRM method was supported by West Virginia NSF Experimental Program to Stimulate Competitive Research (WV-EPSCoR), the Marshall University Center for Cell Differentiation and Development (MU-CDDC) and an NSF award to Simon Collier.

References

  1. 1.
    Gubb, D., and Garcia-Bellido, A. (1982) A genetic analysis of the determination of cuticular polarity during development in Drosophila melanogaster, J Embryol Exp Morphol 68, 37–57.PubMedGoogle Scholar
  2. 2.
    Seifert, J. R., and Mlodzik, M. (2007) Frizzled/PCP signalling: a conserved mechanism regulating cell polarity and directed motility, Nat Rev Genet 8, 126–138.PubMedCrossRefGoogle Scholar
  3. 3.
    Doyle, K., Hogan, J., Lester, M., and Collier, S. (2008) The Frizzled Planar Cell Polarity signaling pathway controls Drosophila wing topography, Dev Biol 317, 354–367.PubMedCrossRefGoogle Scholar
  4. 4.
    Hogan, J., Valentine, M., Cox, C., Doyle, K., and Collier, S. (2011) Two frizzled planar cell polarity signals in the Drosophila wing are differentially organized by the fat/dachsous pathway, PLoS Genet 7, e1001305.PubMedCrossRefGoogle Scholar
  5. 5.
    Valentine, M., and Collier, S. (2011) Planar cell polarity and tissue design: Shaping the Drosophila wing membrane, Fly (Austin) 5(4)PubMedCrossRefGoogle Scholar
  6. 6.
    Kiger, J. A., Jr., Natzle, J. E., Kimbrell, D. A., Paddy, M. R., Kleinhesselink, K., and Green, M. M. (2007) Tissue remodeling during maturation of the Drosophila wing, Dev Biol 301, 178–191.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Biological SciencesMarshall UniversityHuntingtonUSA

Personalised recommendations