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首页> 外文期刊>Investigative ophthalmology & visual science >Retinal cell responses to elevated intraocular pressure: a gene array comparison between the whole retina and retinal ganglion cell layer.
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Retinal cell responses to elevated intraocular pressure: a gene array comparison between the whole retina and retinal ganglion cell layer.

机译：视网膜细胞对眼压升高的反应：整个视网膜和视网膜神经节细胞层之间的基因阵列比较。

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PURPOSE: To determine and compare gene expression patterns in the whole retina and retinal ganglion cell layer (RGCL) in a rodent glaucoma model. METHODS: IOP was unilaterally elevated in Brown Norway rats (N = 26) by injection of hypertonic saline and monitored for 5 weeks. A cDNA microarray was used on whole retinas from one group of eyes with extensive optic nerve injury and on RGCL isolated by laser capture microdissection (LCM) from another group with comparable injury, to determine the significantly up- or downregulated genes and gene categories in both groups. Expression changes of selected genes were examined by quantitative reverse transcription-PCR (qPCR) to verify microarray results. RESULTS: Microarray analysis of the whole retina identified 632 genes with significantly changed expression (335 up, 297 down), associated with 9 upregulated and 3 downregulated biological processes. In contrast, the RGCL microarray yielded 3726 genes with significantly changed expression (2003 up, 1723 down), including 60% of those found in whole retina. Thirteen distinct upregulated biological processes were identified in the RGCL, dominated by protein synthesis. Among 11 downregulated processes, axon extension and dendrite morphogenesis and generation of precursor metabolism and energy were uniquely identified in the RGCL. qPCR confirmed significant changes in 6 selected messages in whole retina and 11 in RGCL. Increased Atf3, the most upregulated gene in the RGCL, was confirmed by immunohistochemistry of RGCs. CONCLUSIONS: Isolation of RGCL by LCM allows a more refined detection of gene response to elevated pressure and improves the potential of determining cellular mechanisms in RGCs and their supporting cells that could be targets for enhancing RGC survival.
机译：目的：确定和比较在啮齿类青光眼模型中整个视网膜和视网膜神经节细胞层（RGCL）中的基因表达模式。方法：通过注射高渗盐水使褐挪威大鼠（N = 26）单眼眼压升高，并监测5周。 cDNA微阵列用于一组患有严重视神经损伤的眼睛的整个视网膜上，以及另一组具有相当损伤的另一组通过激光捕获显微切割术（LCM）分离的RGCL，以确定两者中显着上调或下调的基因和基因类别组。通过定量逆转录PCR（qPCR）检查所选基因的表达变化，以验证微阵列结果。结果：对整个视网膜进行微阵列分析，确定了632个表达明显改变的基因（335个向上，297个向下），与9个上调的生物过程和3个下调的生物过程相关。相比之下，RGCL微阵列产生了3726个基因，其表达发生了显着变化（2003年上升，1723年下降），包括整个视网膜中发现的基因的60％。在RGCL中鉴定出13种不同的上调生物学过程，其中蛋白质合成占主导。在11个下调的过程中，RGCL独特地确定了轴突延伸和树突形态发生以及前体代谢和能量的产生。 qPCR证实了整个视网膜中6条选定信息和RGCL中11条信息的显着变化。通过RGC的免疫组织化学证实Atf3增加，这是RGCL中最上调的基因。结论：通过LCM分离RGCL可以更精确地检测对升高的压力的基因反应，并提高了确定RGC及其支持细胞中细胞机制的潜力，这些机制可能是提高RGC存活率的目标。

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