Light damage induced changes in mouse retinal gene expression
Light damage induced changes in mouse retinal gene expression
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DOI：
10.1016/j.exer.2004.05.002
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发表时间：
2004-08-01
影响因子：
3.4
通讯作者：
Dunaief, JL
中科院分区：
文献类型：
--
作者：
Chen, L;Wu, W;Dunaief, JL
氧化应激在视网膜变性的光损伤模型以及年龄相关性黄斑变性中起作用。本研究的目的是鉴定急性光氧化应激诱导的视网膜基因，这些基因可能作为细胞凋亡的介质或作为存活因子发挥作用。为了实现这一点，将Balb/c小鼠暴露于明亮的冷白色荧光灯下7小时。然后分离视网膜用于总RNA制备，随后进行Affytron DNA微阵列分析，以比较光损伤小鼠与未暴露对照小鼠中的基因表达。进行了三个独立的光损伤实验，并应用统计滤波器来检测平均至少两倍表达变化的基因。进行定量PCR以确认基因表达的改变。70个基因在光损伤后立即上调至少两倍。QPCR证实了所有10个测试基因的上调。上调的基因分为几类，包括抗氧化剂：血浆铜蓝蛋白、金属硫蛋白和血红素加氧酶;抗凋亡基因：bag 3;氯离子通道：clic 1和clic 4;转录因子：c-fos、fra 1、junB、stall、krox-24和c/ebp;分泌的信号分子：几丁质酶3样蛋白I和骨桥蛋白;炎症相关基因：MCP-1和ICAM 1等。五个干扰素-γ应答基因的上调表明光损伤后干扰素水平升高。AP-1转录因子的三个组分的上调与先前的证据相一致，该证据暗示AP-1在光损伤发病机制中。四个铜或铁结合蛋白的上调，表明光氧化应激可能会影响金属稳态。发现的基因上调光损伤可能会影响光感受器的生存受到光氧化应激。(C)2004爱思唯尔有限公司保留所有权利。
Oxidative stress plays a role in the light damage model of retinal degeneration as well as in age-related macular degeneration. The purpose of this study is to identify retinal genes induced by acute photo-oxidative stress, which may function as mediators of apoptosis or as survival factors. To accomplish this, Balb/c mice were exposed to bright cool white fluorescent light for 7 hr. Retinas were then isolated for total RNA preparation followed by Affymetrix DNA microarray analysis to compare gene expression in light damaged mice to unexposed controls. Three independent light damage experiments were carried out and statistical filters were applied to detect genes with expression changes averaging at least two-fold. Quantitative PCR was carried out to confirm altered gene expression. Seventy genes were upregulated at least two-fold immediately following light damage. QPCR confirmed upregulation of all 10 genes tested. The upregulated genes fall into several categories including antioxidants: ceruloplasmin, metallothionein, and heme oxygenase; antiapoptotic gene: bag3, chloride channels: clic1 and clic4; transcription factors: c-fos, fra1, junB, stall, krox-24 and c/ebp; secreted signaling molecules: chitinase 3-like protein I and osteopontin; inflammation related genes: MCP-1 and ICAM1 and others. Upregulation of five interferon-gamma responsive genes suggests elevated interferon levels after light damage. Upregulation of three components of the AP-1 transcription factor is consistent with previous evidence implicating AP-1 in light damage pathogenesis. Four copper or iron binding proteins were upregulated, suggesting that photo-oxidative stress may affect metal homeostasis. The genes found upregulated by light damage may affect the survival of photoreceptors subjected to photo-oxidative stress. (C) 2004 Elsevier Ltd. All rights reserved.