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Methods Mol Biol. 2012;839:91-104. doi: 10.1007/978-1-61779-510-7_8.

Using 32-cell stage Xenopus embryos to probe PCP signaling.

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1
School of Life Sciences, College of Natural Sciences, Kyungpook National University, Daegu, South Korea. leeh@knu.ac.kr

Abstract

Use of loss-of function (via antisense Morpholino oligonucleotides (MOs)) or over-expression of proteins in epithelial cells during early embryogenesis of Xenopus embryos, can be a powerful tool to understand how signaling molecules can affect developmental events. The techniques described here are useful for examining the roles of proteins in cell-cell adhesion, and planar cell polarity (PCP) signaling in cell movement. We describe how to target specific regions within the embryos by injecting an RNA encoding a tracer molecule along with RNA encoding your protein of interest or an antisense MO to knock-down a particular protein within a specific blastomere of the embryo. Effects on cell-cell adhesion, cell movement, and endogenous or exogenous protein localization can be assessed at later stages in specific targeted tissues using fluorescent microscopy and immunolocalization.

PMID:
22218895
PMCID:
PMC4461027
DOI:
10.1007/978-1-61779-510-7_8
[Indexed for MEDLINE]
Free PMC Article
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