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整合素α亚基mRNA在非洲爪蟾早期胚胎中差异表达。

Integrin alpha subunit mRNAs are differentially expressed in early Xenopus embryos.

作者信息

Whittaker C A, DeSimone D W

机构信息

Department of Anatomy and Cell Biology, University of Virginia, Charlottesville 22908.

出版信息

Development. 1993 Apr;117(4):1239-49. doi: 10.1242/dev.117.4.1239.

Abstract

Adhesion of cells to extracellular matrix proteins is mediated, in large part, by transmembrane receptors of the integrin family. The identification of specific integrins expressed in early embryos is an important first step to understanding the roles of these receptors in developmental processes. We have used polymerase chain reaction methods and degenerate oligodeoxynucleotide primers to identify and clone Xenopus integrin alpha subunits from neurula-stage (stage 17) cDNA. Partial cDNAs encoding integrin subunits alpha 2, alpha 3, alpha 4, alpha 5, alpha 6 and an alpha IIb-related subunit were cloned and used to investigate integrin mRNA expression in early embryos by RNase protection assay and whole-mount in situ hybridization methods. Considerable integrin diversity is apparent early in development with integrins alpha 2, alpha 3, alpha 4, alpha 5 and alpha 6 each expressed by the end of gastrulation. Both alpha 3 and alpha 5 are expressed as maternal mRNAs. Zygotic expression of alpha 2, alpha 3, alpha 4 and alpha 6 transcripts begins during gastrulation. Integrin alpha 5 is expressed at relatively high levels during cleavage, blastula and gastrula stages suggesting that it may represent the major integrin expressed in the early embryo. We demonstrated previously that integrin beta 1 protein synthesis remains constant following induction of stage 8 animal cap cells with activin (Smith, J. C., Symes, K., Hynes, R. O. and DeSimone, D. W. (1990) Development 108, 289-298.). Here we report that integrin alpha 3, alpha 4 and alpha 6 mRNA levels increase following induction with 10 U/ml activin-A whereas alpha 5, beta 1 and beta 3 mRNA levels remain unchanged. Whole-mount in situ hybridization reveals that alpha 3 mRNAs are expressed by cells of the involuting mesoderm in the dorsal lip region of early gastrulae. As gastrulation proceeds, alpha 3 expression is localized to a stripe of presumptive notochordal cells along the dorsal midline. In neurulae, alpha 3 mRNA is highly expressed in the notochord but becomes progressively more restricted to the caudalmost portion of this tissue as development proceeds from tailbud to tadpole stages. In addition, alpha 3 is expressed in the forebrain region of later stage embryos. These data suggest that integrin-mediated adhesion may be involved in the process of mesoderm involution at gastrulation and the organization of tissues during embryogenesis.

摘要

细胞与细胞外基质蛋白的黏附在很大程度上是由整合素家族的跨膜受体介导的。鉴定早期胚胎中表达的特定整合素是理解这些受体在发育过程中作用的重要第一步。我们利用聚合酶链反应方法和简并寡脱氧核苷酸引物,从神经胚期(第17阶段)的cDNA中鉴定并克隆非洲爪蟾整合素α亚基。克隆了编码整合素亚基α2、α3、α4、α5、α6和一个αIIb相关亚基的部分cDNA,并通过核糖核酸酶保护分析和整装原位杂交方法研究早期胚胎中整合素mRNA的表达。在发育早期就明显存在相当大的整合素多样性,整合素α2、α3、α4、α5和α6在原肠胚形成结束时均有表达。α3和α5均作为母源mRNA表达。α2、α3、α4和α6转录本的合子表达在原肠胚形成期间开始。整合素α5在卵裂期、囊胚期和原肠胚期表达水平相对较高,这表明它可能是早期胚胎中表达的主要整合素。我们先前证明,用激活素诱导8期动物帽细胞后,整合素β1蛋白合成保持恒定(史密斯,J.C.,西姆斯,K.,海因斯,R.O.和德西蒙内,D.W.(1990年)《发育》108,289 - 298)。在此我们报告,用10 U/ml激活素 - A诱导后,整合素α3、α4和α6的mRNA水平升高,而α5、β1和β3的mRNA水平保持不变。整装原位杂交显示,α3 mRNA在早期原肠胚背唇区域内卷中胚层的细胞中表达。随着原肠胚形成的进行,α3表达定位于沿背中线的一条假定脊索细胞带。在神经胚中,α3 mRNA在脊索中高度表达,但随着发育从尾芽期到蝌蚪期进行,其表达逐渐更局限于该组织的最尾端部分。此外,α3在后期胚胎的前脑区域表达。这些数据表明,整合素介导的黏附可能参与原肠胚形成时中胚层的内卷过程以及胚胎发育期间组织的组织化过程。

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