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Analyses of cell recognition mechanisms by genetic manipulation

通过基因操作分析细胞识别机制

基本信息

批准号：
63440005
负责人：
TAKEICHI Masatoshi
金额：
$ 14.72万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1991
项目状态：
已结题

项目摘要

We obtained the following results : 1. We searched for novel. cadherins, and identified R-cadherin. This cadherin was mainly expressed in the nervous system, but its expression occured at later developmental stages than Ncadherin. The distribution patterns of these cadherins were also distinct. 2. By PCR using brain MRNA as template, we obtained other novel cadherin CDNA fragments. We cloned two of them and determined their sequences. Cadherins encoded by these cDNAs showed some unique features which demarcate them from classical cadherins. These results suggest that the cadherin family should be divided. into subfamilies. 3. We attempted to identify Drosophila cadherins using PCR', and could obtain CDNA fragments which can be divided into four groups. Also, we could clone -a Drosophila homologue of vertebrate (x-catenin. 4. To investigate the morphogenetic role of cadherins, we injected N-cadherin mRNA into Xenopus 2-cell stage embryos. The overexpression of N-cadherin in embryos induced various morphological abnormalities, including thickening of epidermis and separation of cell masses. The ectopic expression of N-cadherin also affected some morphogenetic movements. These findings confirmed the importance of. regulated expression of cadherins in morphogenesis. 5. We demonstrated that catenins, cadherin-associated proteins, are crucial for cadherin function. We also cloned cc-catenin, and found that it is similar to vinculin.

我们得到了以下结果：1.我们搜索小说。钙粘蛋白和鉴定的R-钙粘蛋白。这种钙粘蛋白主要在神经系统中表达，但其表达发生在比Ncadherin晚的发育阶段。这些钙粘蛋白的分布模式也是不同的。2.以脑组织mRNA为模板，通过PCR扩增，获得了其它新的钙粘蛋白cDNA片段。我们克隆了其中两个并确定了它们的序列。由这些cDNA编码的钙粘蛋白显示出一些独特的特征，这些特征将它们与经典的钙粘蛋白区分开来。这些结果表明，钙粘蛋白的家庭应该分开。分为亚家族3.我们尝试用PCR方法鉴定果蝇钙粘蛋白，获得了四组cDNA片段。此外，我们可以克隆-一个果蝇同源的脊椎动物（x-连环蛋白。4.为了研究钙粘蛋白的形态发生作用，我们将N-钙粘蛋白mRNA注射到爪蟾2细胞期胚胎中。N-钙粘蛋白在胚胎中的过度表达诱导了各种形态学异常，包括表皮增厚和细胞团分离。异位表达的N-钙粘蛋白也影响一些形态发生的运动。这些发现证实了。钙粘蛋白在形态发生中的调节表达。5.我们证明，连环蛋白，钙粘蛋白相关蛋白，是至关重要的钙粘蛋白的功能。我们还克隆了α-连环蛋白，发现它与黏着斑蛋白相似。