XB-ART-44653Methods Mol Biol January 1, 2012; 839 91-104.
Using 32-cell stage Xenopus embryos to probe PCP signaling.
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.
PubMed ID: 22218895
PMC ID: PMC4461027
Article link: Methods Mol Biol
Species referenced: Xenopus
Genes referenced: daam1
Morpholinos: daam1 MO1
References [+] :
Bauer, The cleavage stage origin of Spemann's Organizer: analysis of the movements of blastomere clones before and during gastrulation in Xenopus. 1994, Pubmed, Xenbase