Analyzing Planar Cell Polarity During Zebrafish Gastrulation
Planar cell polarity was first described in invertebrates over 20 years ago and is defined as the polarity of cells (and cell structures) within the plane of a tissue, such as an epithelium. Studies in the last 10 years have identified critical roles for vertebrate homologs of these planar cell polarity proteins during gastrulation cell movements. In zebrafish, the terms convergence and extension are used to describe the collection of morphogenetic movements and cell behaviors that contribute to narrowing and elongation of the embryonic body plan. Disruption of planar cell polarity gene function causes profound defects in convergence and extension creating an embryo that has a shortened anterior–posterior axis and is broadened mediolaterally. The zebrafish gastrula-stage embryo is transparent and amenable to live imaging using both Nomarski/differential interference contrast and fluorescence microscopy. This chapter describes methods to analyze convergence and extension movements at the cellular level and thereby connect embryonic phenotypes with underlying planar cell polarity defects in migrating cells.
Key wordsConvergence and extension Ectoderm Gastrulation Nomarski/DIC Mesoderm Planar cell polarity Van gogh-like 2 Wnt Zebrafish
I thank my colleagues Diane Sepich and Lila Solnica-Krezel for their investment in my training in all methods concerning zebrafish gastrulation. Work in the Jessen lab is supported by grants from the American Cancer Society (RSG-09-281-01 DDC) and National Science Foundation (IOS 0950849).
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