Berger Corinna D, März Martin, Kitzing Thomas M, Grosse Robert, Steinbeisser Herbert
Institute of Human Genetics, Section Developmental Genetics, University Heidelberg, Heidelberg, Germany.
Dev Dyn. 2009 Jun;238(6):1407-11. doi: 10.1002/dvdy.21884.
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. (c) 2009 Wiley-Liss, Inc.
Rho家族的小GTP酶是细胞骨架的重要调节因子，在胚胎发育过程中调节细胞形态发生运动。在非洲爪蟾胚胎中，Rho信号传导有助于调节原肠胚和神经胚阶段的汇聚延伸（CE）运动以及组织分离（TS）。在这里，我们描述了一种方法，可用于检测固定的非洲爪蟾组织中活化的（GTP结合的）Rho。该检测方法利用了在细菌中产生并可通过生化方法纯化的Rhotekin与绿色荧光蛋白的融合蛋白（RBD-GFP）。这项技术能够对发育中的胚胎中的Rho信号进行时空分析。《发育动力学》238:1407 - 1411, 2009年。(c) 2009威利 - 利斯公司。
