Gene microarray analysis of experimental glaucomatous retina from cynomologous monkey
Gene microarray analysis of experimental glaucomatous retina from cynomologous monkey
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DOI：
10.1167/iovs.02-1032
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发表时间：
2003-10-01
影响因子：
4.4
通讯作者：
Yoshimura, N
中科院分区：
文献类型：
--
作者：
Miyahara, T;Kikuchi, T;Yoshimura, N
目的。为系统探讨激光诱导的青光眼猴视网膜基因表达的变化，验证实验性青光眼的基因芯片数据。方法：4只猴的右眼采用重复激光小梁网光凝诱导青光眼。左眼作为对照。分别于光凝后30d取出青光眼和对照眼的视网膜。利用人类微阵列芯片分析基因表达的变化，共显示9182个元素，包括表达序列标签(EST)克隆。实时定量聚合酶链式反应进一步证实了部分基因表达的变化。免疫组织化学研究也检测了一些基因产物的蛋白表达，这些基因通过微阵列分析显示上调或下调。结果：产生两只轻度青光眼和两只重度青光眼。在轻度和重度青光眼视网膜中，上调基因分别为45和18个，下调基因分别为17和21个。上调或下调的基因数量占所有检测基因的0.7%。实时定量聚合酶链式反应分析证实了芯片分析中发现的一些基因的表达变化。铜蓝蛋白是上调基因之一，免疫组织化学分析发现铜蓝蛋白在Muller细胞中表达。结论：检测了激光诱导的青光眼猴视网膜中的基因表达谱，只有极少数基因在青光眼中表达上调或下调。在Muller细胞中发现铜蓝蛋白表达上调。
PURPOSE. To systematically explore changes in gene expression in the retina of monkeys with laser-induced glaucoma and to validate the microarray data on eyes with experimental glaucoma.METHODS. Glaucoma was induced in the right eye of four monkeys by repeated argon laser photocoagulation of the trabecular meshwork. The left eye served as the control. Retinas were isolated from glaucomatous and control eyes 30 days after photocoagulation. Gene expression changes were analyzed by human microarray chips which displayed a total of 9182 elements including Expression Sequence Tag (EST) clones. Changes in the expression of some genes were further confirmed by real-time PCR analysis. Immunohistochemical studies to examine protein expression of some gene products were also done for several genes that showed up- or downregulation by the microarray analysis.RESULTS. Two eyes with mild glaucoma and two with severe glaucoma were produced. In the mild and severe glaucomatous retina, the number of upregulated genes was 45 and 18, and the number of downregulated genes was 17 and 21, respectively. The number of genes that were up- or downregulated was 0.7% of all the genes examined. The real-time PCR analysis confirmed expression changes of some genes found in the microarray analysis. Ceruloplasmin was one of the upregulated genes, and it was found by immunohistochemical analyses to be expressed in Muller cells.CONCLUSIONS. Gene expression profiles in laser-induced glaucomatous monkey retinas were determined, and only a very small population of genes was up- or downregulated in glaucomatous eyes. Upregulation of ceruloplasmin protein was found in the Muller cells.