Malosio M L, Gilardelli D, Paris S, Albertinazzi C, de Curtis I
Cell Adhesion Unit, Department of Biological and Technological Research (DIBIT), San Raffaele Scientific Institute, 20132 Milano, Italy.
J Neurosci. 1997 Sep 1;17(17):6717-28. doi: 10.1523/JNEUROSCI.17-17-06717.1997.
Previous studies on small GTP-binding proteins of the Rho family have revealed their involvement in the organization of cell actin cytoskeleton. The function of these GTPases during vertebrate development is not known. With the aim of understanding the possible role of these proteins during neuronal development, we have cloned and sequenced five members expressed in developing chick neural retinal cells. We have identified four chicken genes, cRhoA, cRhoB, cRhoC, and cRac1A, homologous to known human genes, and a novel Rac gene, cRac1B. Analysis of the distribution of four of the identified transcripts in chicken embryos shows for the first time high levels of expression of Rho family genes in the vertebrate developing nervous system, with distinct patterns of distribution for the different transcripts. In particular, cRhoA and cRac1A gene expression appeared ubiquitous in the whole embryo, and the cRhoB transcript was more prominent in populations of neurons actively extending neurites, whereas the newly identified cRac1B gene was homogeneously expressed only in the developing nervous system. Temporal analysis of the expression of the five genes suggests a correlation with the morphogenetic events occurring within the developing retina and the retinotectal pathway. Expression of an epitope-tagged cRac1B in retinal neurons showed a diffuse distribution of the protein in the cell body and along neurites. Taken as a whole, our results suggest important roles for ubiquitous and neural-specific members of the Rho family in the acquisition of the mature neuronal phenotype.
先前对Rho家族小GTP结合蛋白的研究表明,它们参与细胞肌动蛋白细胞骨架的组织。这些GTP酶在脊椎动物发育过程中的功能尚不清楚。为了了解这些蛋白在神经元发育过程中可能发挥的作用,我们克隆并测序了在发育中的鸡神经视网膜细胞中表达的五个成员。我们鉴定出四个与已知人类基因同源的鸡基因,即cRhoA、cRhoB、cRhoC和cRac1A,以及一个新的Rac基因cRac1B。对鸡胚胎中四个已鉴定转录本分布的分析首次表明,Rho家族基因在脊椎动物发育中的神经系统中高水平表达,不同转录本具有不同的分布模式。特别是,cRhoA和cRac1A基因表达在整个胚胎中普遍存在,cRhoB转录本在积极延伸神经突的神经元群体中更为突出,而新鉴定的cRac1B基因仅在发育中的神经系统中均匀表达。对这五个基因表达的时间分析表明,它们与发育中的视网膜和视网膜顶盖通路中发生的形态发生事件相关。在视网膜神经元中表达表位标记的cRac1B表明,该蛋白在细胞体和神经突中呈弥散分布。总体而言,我们的结果表明,Rho家族中普遍存在的成员和神经特异性成员在获得成熟神经元表型中发挥着重要作用。