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DNA microarray analysis of the gene expression changes in retinal neuronal cell death.

DNA微阵列分析视网膜神经细胞死亡中基因表达的变化。

基本信息

批准号：
13470364
负责人：
YOSHIMURA Nagahisa
金额：
$ 10.37万
依托单位：
Shinshu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470364/
关键词：
Neuronal cell death Retinal ischemia-reperfusion Glaucoma DNA microarray Gene expression ラット網膜虚血再灌流障害サル緑内障モデル gene chip 遺伝子発現 real time PCR 網膜神経細胞死 DNAマイクロチップ

项目摘要

Two experimental models were used in this study. The first model was rat retinal is3chemia-reperfusion model. In this model, gene expression changes were systematically studied by using DNA microarrays that displayed more than 10,000 genes and ESTs. The number of genes that showed up-regulation of more than 3 folds was as many as 135 genes and such genes can be grouped into 6 different groups. (1) transcription factors (2) cell cycle-related genes (3) stress responsive genes (4) cell signaling genes (5) cell adhesion molecule genes (6) protease genes. Expression changes of some genes that belongs (1),(2),and (3) were also studied by the real time PCR analysis. In this model, importance of transcription factors and cell cycle-related genes was shown. The second model used was monkey glaucoma model. Ocular hypertension was induced in monkey eyes by laser photocoagulation of the trabecular meshwork. In this model, gene expression changes in the sensory retina were studied by using a human DNA microarray because ready-made monkey microarrays were not available. Contrary to the rat retinal ischemia-reperfusion model, the profile of gene expression changes found in the monkey glaucoma model was different from that found in the rat ischemia-reperfusion model. Only 0.7% of the genes studied showed up-regulation of more than 1.7 folds of the control. Among them, ceruloplasmin showed up-regulation and the expression changes were validated by the real time PCR, analysis and immunohistochemical studies. Three peer-reviewed papers have been published through the present study.

本研究中使用了两种实验模型。第一种模型为大鼠视网膜缺血再灌注模型。在该模型中，通过使用显示超过10，000个基因和EST的DNA微阵列系统地研究了基因表达变化。表达上调3倍以上的基因多达135个，这些基因可分为6个不同的组。(1)转录因子（2）细胞周期相关基因（3）应激反应基因（4）细胞信号传导基因（5）细胞粘附分子基因（6）蛋白酶基因。通过真实的实时PCR分析了（1）、（2）和（3）中部分基因的表达变化。在该模型中，转录因子和细胞周期相关基因的重要性被证明。第二种模型为猴青光眼模型。通过激光光凝小梁网在猴眼中诱导高眼压。在该模型中，由于没有现成的猴子微阵列，因此通过使用人类DNA微阵列研究了感觉视网膜中的基因表达变化。与大鼠视网膜缺血-再灌注模型相反，在猴青光眼模型中发现的基因表达变化的概况与在大鼠缺血-再灌注模型中发现的不同。只有0.7%的基因表达上调超过对照组的1.7倍。其中铜蓝蛋白表达上调，真实的时间PCR分析和免疫组化研究证实了其表达变化。通过本研究已发表了三篇同行评审论文。