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Detection of activated Rho in fixed Xenopus tissue
Author(s) -
Berger Corinna D.,
März Martin,
Kitzing Thomas M.,
Grosse Robert,
Steinbeisser Herbert
Publication year - 2009
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.21884
Subject(s) - xenopus , biology , neurula , convergent extension , gastrulation , microbiology and biotechnology , embryo , green fluorescent protein , gtpase , cytoskeleton , embryogenesis , cell , genetics , gene
Small GTPases of the Rho family are important modulators of the cytoskeleton and regulate morphogenetic cell movements during embryonic development. In the Xenopus embryo, Rho signaling contributes to the regulation of convergent extension (CE) movements in gastrula and neurula stages as well as to tissue separation (TS). Here we describe a method that allows the detection of activated (GTP‐bound) Rho in fixed Xenopus tissue. The assay makes use of a fusion protein of Rhotekin and Green‐Fluorescent‐Protein (RBD‐GFP), which is produced in bacteria and can be purified biochemically. This technique allows a temporal and spatial analysis of Rho signaling in the developing embryo. Developmental Dynamics 238:1407–1411, 2009. © 2009 Wiley‐Liss, Inc.