Convergent Extension Analysis in Mouse Whole Embryo Culture

  • Sophie E. Pryor
  • Valentina Massa
  • Dawn Savery
  • Nicholas D. E. Greene
  • Andrew J. Copp
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 839)

Abstract

Mutations have been identified in a non-canonical Wnt signalling cascade (the planar cell polarity pathway) in several mouse genetic models of severe neural tube defects. In each of these models, neurulation fails to be initiated at the 3–4 somite stage, leading to an almost entirely open neural tube (termed craniorachischisis). Studies in whole embryo culture have identified a defect in the morphogenetic process of convergent extension during gastrulation, preceding the onset of neural tube closure. The principal defect is a failure of midline extension, both in the neural plate and axial mesoderm. This leads to an abnormally wide neural plate in which the elevating neural folds are too far apart to achieve closure. In this chapter, we provide details of several experimental methods that can be used to evaluate convergent extension in cultured mouse embryos. We describe analytical methods that can reveal the abnormalities that characterise neurulation-stage embryos with defective planar cell polarity signalling, in particular the loop-tail (Lp; Vangl2) mutant.

Key words

Mouse Neurulation Whole embryo culture DiI labelling Electroporation Convergent extension Neural tube defects Craniorachischisis Vangl2 Loop-tail 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Sophie E. Pryor
    • 1
  • Valentina Massa
    • 2
  • Dawn Savery
    • 1
  • Nicholas D. E. Greene
    • 1
  • Andrew J. Copp
    • 1
  1. 1.Neural Development Unit, Institute of Child HealthUniversity College LondonLondonUK
  2. 2.Dulbecco Telethon Institute at M.Tettamanti Research Center, Pediatric DepartmentUniversity of Milano-BicoccaMonzaItaly

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