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Functional analysis of human corneal endothelial cells based on the cDNA library

基于cDNA文库的人角膜内皮细胞功能分析

基本信息

批准号：
16390494
负责人：
YAMAGAMI Satoru
金额：
$ 4.99万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390494/
关键词：
cDNA corneal endothelium matrix gla protein DDB2 ヒト角膜内皮細胞遺伝子修復

项目摘要

The gene expression profile of human corneal endothelium (CE) was established with the gene signature system. A novel gene, GS3582, was abundantly transcribed gene in the CE compared with other tissues using a human gene expression database. To investigate the physiological function of CESP-1, we assessed its tissue distribution and subcellular localization in humans and mice. The rabbit and mouse CESP-1 cDNA sequences contained an open reading frame coding 242 and 283 amino acids, respectively. Mouse CESP-1 is entirely consistent with mouse OSAP. CESP-1 was expressed in the human corneal epithelium, CE, cultured CE, brain, testis, and ovary on western blotting analysis. Mouse CESP-1 was also expressed in mouse corneal epithelium and CE with anti-mouse C- but not N--terminal OSAP Ab according to immunohistochemical analysis. Subcellular localization of CESP-1 to the mitochondria was demonstrated in cultured human CE. The N-terminal of CESP-1, possessing a mitochondrial targeting sequence, may be processed after the protein is imported into the mitochondria. CESP-1 was distributed in the corneal epithelium, the CE and cultured human CE, as well as the brain, testis, and ovary. CESP-1 was localized in the mitochondria of cultured human CE. These findings may provide some clues about the physiological function of CESP-1.

利用基因标记系统建立了人角膜内皮（CE）的基因表达谱。一个新的基因GS3582在CE中被大量转录。为了研究csp -1的生理功能，我们评估了它在人和小鼠中的组织分布和亚细胞定位。家兔和小鼠csp -1 cDNA序列包含一个开放阅读框，分别编码242个和283个氨基酸。小鼠csp -1与小鼠OSAP完全一致。western blotting分析显示，csp -1在人角膜上皮、CE、培养CE、脑、睾丸和卵巢中均有表达。免疫组化分析显示，小鼠csp -1在小鼠角膜上皮和CE中也有表达，而抗小鼠C端OSAP Ab不表达。在培养的人CE中证实了csp -1在线粒体的亚细胞定位。csp -1的n端具有线粒体靶向序列，在该蛋白被导入线粒体后可能会被加工。csp -1分布于角膜上皮、CE和培养的人CE，以及脑、睾丸和卵巢。csp -1定位于培养的人CE的线粒体中。这些发现可能为了解csp -1的生理功能提供一些线索。