Time course analysis of gene expression during light-induced photoreceptor cell death and regeneration in albino zebrafish
Time course analysis of gene expression during light-induced photoreceptor cell death and regeneration in albino zebrafish
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DOI：
10.1002/dneu.20362
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发表时间：
2007-07-01
影响因子：
3
通讯作者：
Hyde, David R.
中科院分区：
文献类型：
--
作者：
Kassen, Sean C.;Ramanan, Vijay;Hyde, David R.
恒定强光引起成年白化病斑马视杆细胞和视锥细胞凋亡。光感受器随后从增殖的内核层（INL）祖细胞再生，所述祖细胞迁移到外核层（ONL）并分化成视杆细胞和视锥细胞。为了鉴定在光感受器再生反应期间的基因表达变化，在光处理期间的五个时间点进行微阵列分析。时间过程包括感光细胞死亡期间的早期时间点（16 h）、祖细胞增殖和迁移期间的晚期时间点（31、51和68 h）和96 h时间点，这可能对应于初始感光细胞分化。在每个时间点相对于对照（0小时光照）计算每个基因的平均表达值，并且通过单因素ANOVA的统计分析鉴定出4567个基因，其显示基因表达沿着时间过程的显著变化。该数据集中的基因根据其时间表达模式和建议功能进行聚类。实时定量PCR验证了选定基因的微阵列表达谱，包括stat3，其表达在光照过程中显着增加。基于免疫印迹，总的和活化的Stat3蛋白表达也增加在光处理。Stat3在视网膜组织切片上的免疫定位表明，通过16小时的光暴露，光感受器和Muller神经胶质中的表达增加。一些Stat3阳性的Muller细胞在31 h时表达PCNA，表明Stat3可能在再生反应过程中信号传导Muller细胞亚群增殖中发挥作用。(c)2007 Wiley Periodicals，Inc.
Constant intense light causes apoptosis of rod and cone photoreceptors in adult albino zebralish. The photoreceptors subsequently regenerate from proliferating inner nuclear layer (INL) progenitor cells that migrate to the outer nuclear layer (ONL) and differentiate into rods and cones. To identify gene expression changes during this photoreceptor regeneration response, a microarray analysis was performed at five time points during the light treatment. The time course included an early time point during photoreceptor death (16 h), later time points during progenitor cell proliferation and migration (31, 51, and 68 h) and a 96 h time point, which likely corresponds to the initial photoreceptur differentiation. Mean expression values for each gene were calculated at each time point relative to the control (0 h light exposure) and statistical analysis by one-way ANOVA identified 4567 genes exhibiting significant changes in gene expres-sion along the time course. The genes within this data set were clustered based on their temporal expression patterns and proposed functions. Quantitative real-time PCR validated the microarray expression profiles for selected genes, including stat3 whose expression increased markedly during the light exposure. Based on immunoblots, both total and activated Stat3 protein expression also increased during the light treatment. Immunolocalization of Stat3 on retinal tissue sections demonstrated increased expression in photoreceptors and Muller glia by 16 h of light exposure. Some of the Stat3-positive Muller cells expressed PCNA at 31 h, suggesting that Stat3 may play a role in signaling a subset of Muller cells to proliferate during the regeneration response. (c) 2007 Wiley Periodicals, Inc.