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

本研究使用了两个实验模型。第一种模型为大鼠视网膜缺血再灌注模型。在这个模型中，利用显示了10,000多个基因和EST的DNA微阵列系统地研究了基因表达的变化。上调3倍以上的基因多达135个，可分为6个不同的类群。(1)转录因子(2)细胞周期相关基因(3)应激反应基因(4)细胞信号基因(5)细胞黏附分子基因(6)蛋白酶基因。利用实时荧光定量聚合酶链式反应技术研究了(1)、(2)、(3)基因的表达变化。在这个模型中，显示了转录因子和细胞周期相关基因的重要性。第二种模型为猴青光眼模型。用激光小梁网光凝法诱导猴眼高眼压。在这个模型中，由于没有现成的猴子微阵列，因此使用人类DNA微阵列来研究感觉性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，猴青光眼模型的基因表达变化与大鼠缺血再灌注模型不同。在所研究的基因中，只有0.7%的基因表达上调超过对照的1.7倍。其中，铜蓝蛋白表达上调，并通过实时荧光定量聚合酶链式反应、分析和免疫组织化学研究证实其表达变化。本研究发表了三篇同行评议的论文。

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.