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Gene Loss-of-Function and Live Imaging in Chick Embryos

  • Anne C. Rios
  • Christophe MarcelleEmail author
  • Olivier Serralbo
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 839)

Abstract

Planar cell polarity (PCP) is the coordinate organization of cells within the plane of a tissue. PCP is essential for tissue function, such as for proper hearing in the vertebrate ear or for accurate vision in the Drosophila eye. Using the chick embryo, we have recently shown that during early muscle formation, the first formed muscle fibres utilize the PCP pathway to orient parallel to a WNT11 source present in the medial border of somites. Our results further establish that WNT11 acts as a directional cue to regulate this process. To perform this study, two major techniques have been utilized, the gene loss-of-function using a vector-based shRNAmir expression and confocal videomicroscopy of fluorescent gene reporters targeted in specific cell subpopulations by in vivo electroporation. Here we describe the two techniques.

Key words

Chick embryo Somite shRNAmir Confocal videomicroscopy In vivo imaging 

Notes

Acknowledgments

This study was funded by the Agence nationale de la recherche (ANR) and the EU sixth Framework Programme Network of Excellence MYORES. Martin Scaal and Jerome Gros have initiated the in vivo video microscopy. The help of Pascal Weber, Stephen Firth, and Chad Johnson from Imaging Facilities (IBDML, Marseille and MMI, Monash University) is acknowledged.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Anne C. Rios
    • 1
  • Christophe Marcelle
    • 1
    Email author
  • Olivier Serralbo
    • 1
  1. 1.EMBL Australia, Australian Regenerative Medicine Institute (ARMI)Monash UniversityClaytonAustralia

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