Protocol

Planar Cell Polarity

Volume 839 of the series Methods in Molecular Biology pp 133-146

Date:

Convergent Extension Analysis in Mouse Whole Embryo Culture

  • Sophie E. PryorAffiliated withNeural Development Unit, Institute of Child Health, University College London
  • , Valentina MassaAffiliated withDulbecco Telethon Institute at M.Tettamanti Research Center, Pediatric Department, University of Milano-Bicocca
  • , Dawn SaveryAffiliated withNeural Development Unit, Institute of Child Health, University College London
  • , Nicholas D. E. GreeneAffiliated withNeural Development Unit, Institute of Child Health, University College London
  • , Andrew J. CoppAffiliated withNeural Development Unit, Institute of Child Health, University College London Email author 

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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