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Role of the ret proto-oncogene in the development of the enteric nervous system

ret原癌基因在肠神经系统发育中的作用

基本信息

批准号：
06454192
负责人：
TAKAHASHI Masahide
金额：
$ 4.22万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454192/
关键词：
ret protooncogene tyrosine kinase Hirschsprung's disease enteric neuron Hirschosprung病腸管神経系

项目摘要

Immunohistochemical analysis with the anti-Ret antibody was performed to investigate the expression of the c-ret proto-oncogene product (c-Ret protein) in embryonic, and adult rat tissues. During embryogenesis, it was expressed at high levels in the enteric neuroblasts and the sympathetic ganglia. c-Ret positive cells appeared in the mesenchyme around the rofegut and the dorsal aorta at day 11.5 and formed the myeteric plexus of the whole embryonic gut and the sympathetic truck at later stages, respectively. In the enteric nervous system, c-Ret positive cells began to contact each other to form the myenteric ganglia and were arranged between the muscle layrs. After birth, these neurons continued to express the c-Ret protein at variable levels. These results suggested that the c-Ret protein might be involved in the development of the enteric and sympathetic nervous system.We introduced several mutations of the ret proto-oncogene found in the Hirchsprung's disease.ret cDNA with each mutation was transfected into NIH 3T3 cells, cell lines expressing the mutant Ret proteins were established. The Ret proteins were usually expressed as a mature glycosylated form of 170 kDa protein present on the cell surface and an immature glycosylated form of the 150 kDa Ret protein in the endoplasmic reticulum. Western blot analysis revealed that very little of the 170 kDa Ret protein was expressed in transfectants while expression of the 150 kDa Ret protein was not affected. This result suggested that the Hirschsprung mutations impair the transport of the Ret protein to the plasma membrane.

用抗Ret抗体进行免疫组织化学分析，以研究胚胎和成年大鼠组织中c-Ret原癌基因产物（c-Ret蛋白）的表达。在胚胎发育过程中，它在肠神经母细胞和交感神经节中高水平表达。c-Ret阳性细胞于11.5d出现在小肠和背主动脉周围的间充质中，后期分别形成整个胚胎肠的肠系膜丛和交感神经干。在肠神经系统中，c-Ret阳性细胞开始相互接触，形成肌间神经节，并排列在肌层之间。出生后，这些神经元继续以不同的水平表达c-Ret蛋白。本研究将在先天性巨结肠病中发现的ret原癌基因的几个突变点导入NIH 3 T3细胞，建立了表达突变Ret蛋白的细胞系。Ret蛋白通常表达为存在于细胞表面上的170 kDa蛋白的成熟糖基化形式和存在于内质网中的150 kDa Ret蛋白的未成熟糖基化形式。Western印迹分析表明，非常少的170 kDa的Ret蛋白表达的转染子，而表达的150 kDa的Ret蛋白不受影响。该结果表明，先天性巨结肠突变损害了Ret蛋白向质膜的转运。