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.
School of Life Sciences, College of Natural Sciences, Kyungpook National University, Daegu, South Korea. leeh@knu.ac.kr
Hyun-Shik
HS
Lee
Sergei Y
SY
Sokol
Sally A
SA
Moody
Ira O
IO
Daar
D000818:Animals; D001757:Blastomeres / Q000166:cytology; D016764:Cell Polarity; D004625:Embryo, Nonmammalian / Q000166:cytology*; D005260:Female; D005455:Fluorescent Antibody Technique; D008297:Male; D015398:Signal Transduction; D016610:Tissue Embedding; D016707:Tissue Fixation; D014981:Xenopus / Q000196:embryology*; D001616:beta-Galactosidase / Q000378:metabolism
D001616:beta-Galactosidase