Microarray Analysis of XOPS-mCFP Zebrafish Retina Identifies Genes Associated with Rod Photoreceptor Degeneration and Regeneration
Microarray Analysis of XOPS-mCFP Zebrafish Retina Identifies Genes Associated with Rod Photoreceptor Degeneration and Regeneration
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DOI：
10.1167/iovs.10-6022
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发表时间：
2011-04-01
影响因子：
4.4
通讯作者：
Fadool, James M.
中科院分区：
文献类型：
--
作者：
Morris, Ann C.;Forbes-Osborne, Marie A.;Fadool, James M.
目的。 XOPS-mCFP 转基因斑马鱼在一生中经历视杆细胞发育和退化的连续循环，使它们成为研究视杆细胞光感受器再生的分子决定因素的有用模型。本研究的目的是比较野生型和 XOPS-mCFP 视网膜的基因表达谱，并鉴定可能有助于视杆细胞再生的基因。对成人野生型和 XOPS-mCFP 视网膜 mRNA 进行微阵列分析。路径分析用于识别数据集中显着代表的生物学相关过程。通过RT-PCR验证表达变化。通过原位杂交和免疫组织化学以及在转基因荧光报告系中进一步检查视网膜发育期间和成人视网膜中选定的基因。 结果。与野生型对照中的表达相比，超过 600 个基因在 XOPS-mCFP 视网膜中表现出显着的表达变化。许多下调的基因与光转导相关，而上调的基因与多种生物功能相关，包括细胞周期、DNA 复制和修复以及细胞发育和死亡。对这些基因的一部分进行 RT-PCR 分析证实了微阵列结果。三种转录因子（sox11b、insm1a 和 c-myb）在 XOPS-mCFP 视网膜中表达增加，在整个视网膜发育过程中和持续神经源性睫状边缘区中也表达。结论。这项研究发现了斑马鱼视杆细胞退化引起的许多基因表达变化，并进一步表明转录调节因子 sox11b、insm1a 和 c-myb 在视网膜发育和视杆细胞光感受器再生中的作用。 （投资眼科可见科学。2011；52：2255-2266）DOI：10.1167/iovs.10-6022
PURPOSE. XOPS-mCFP transgenic zebrafish experience a continual cycle of rod photoreceptor development and degeneration throughout life, making them a useful model for investigating the molecular determinants of rod photoreceptor regeneration. The purpose of this study was to compare the gene expression profiles of wild-type and XOPS-mCFP retinas and identify genes that may contribute to the regeneration of the rods.METHODS. Adult wild-type and XOPS-mCFP retinal mRNA was subjected to microarray analysis. Pathway analysis was used to identify biologically relevant processes that were significantly represented in the dataset. Expression changes were verified by RT-PCR. Selected genes were further examined during retinal development and in adult retinas by in situ hybridization and immunohistochemistry and in a transgenic fluorescent reporter line.RESULTS. More than 600 genes displayed significant expression changes in XOPS-mCFP retinas compared with expression in wild-type controls. Many of the downregulated genes were associated with phototransduction, whereas upregulated genes were associated with several biological functions, including cell cycle, DNA replication and repair, and cell development and death. RT-PCR analysis of a subset of these genes confirmed the microarray results. Three transcription factors (sox11b, insm1a, and c-myb), displaying increased expression in XOPS-mCFP retinas, were also expressed throughout retinal development and in the persistently neurogenic ciliary marginal zone.CONCLUSIONS. This study identified numerous gene expression changes in response to rod degeneration in zebrafish and further suggests a role for the transcriptional regulators sox11b, insm1a, and c-myb in both retinal development and rod photoreceptor regeneration. (Invest Ophthalmol Vis Sci. 2011;52:2255-2266) DOI:10.1167/iovs.10-6022