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视网膜是视觉的基础, 结构十分精细复杂, 视网膜的功能完全依赖于视网膜的结构. 鉴于目前人们对视网膜发生的调控基因和机理了解很少, 利用含16361个基因的芯片分别检测了12~16周、22~26周胎儿及20~40岁成人视网膜中基因的表达水平, 发现814个基因在1或2个时间点表达水平相差3倍以上, 其中表达强度值在1个以上的时间点超过100的差异表达基因共106个, 依次是发育、分化、信号传导、蛋白质合成翻译、代谢、DNA合成修复重组等相关基因. 基因表达模式和聚类分析揭示随视网膜发育成熟呈下调趋势的基因最多, 而呈上调趋势的基因较少. 在上述106个差异表达基因中, 有46个目前在美国国立卫生院(NIH)眼科研究所(NEI)的视网膜cDNA或EST数据库中尚找不到, 它们在视网膜中的作用和功能也不清楚. 为了进一步确定基因芯片结果的可靠性, 采用荧光定量 RT-PCR和常规RT-PCR检测分析了上述46个差异表达基因及另外6个已知的视网膜特异表达基因的表达量, 其中27个基因的表达谱与芯片检测结果完全一致. 另外, 还采用原位杂交技术检测了NNAT基因在视网膜内的表达量及表达产物的细胞和亚细胞分布. 此外, 对106个差异表达基因的染色体定位进行检索揭示, 其中1个基因位于已知的视网膜锥体或锥-杆体营养不良基因位点处, 并与该病的
Retina is the basis of vision, the structure is very complex and complex, the function of the retina is completely dependent on the structure of the retina.At present, people have little understanding of the regulatory genes and mechanisms of retinal development, using 16361 gene chips were detected for 12 to 16 weeks , 22-26 weeks fetus and 20 ~ 40 years old adult retina, we found that the expression level of 814 genes was more than 3 times in one or two time points, in which the expression intensity value in more than one time point exceeded 100 Of which 106 genes were differentially expressed, followed by genes related to development, differentiation, signal transduction, protein synthesis, translation, metabolism, DNA synthesis, repair and recombination, etc. Gene expression patterns and cluster analysis revealed that down- While the up-regulated genes were less.46 Of the 106 differentially expressed genes mentioned above, 46 are currently not found in the retinal cDNA or EST database of the National Institutes ofHeaerhomes Eye (NEI) The role and function of the retina is not clear.In order to further determine the reliability of gene chip results, using quantitative RT-PCR and conventional RT-PCR The expression levels of the above 46 differentially expressed genes and the other 6 known retina-specific genes were analyzed and assayed, and the expression profiles of the 27 genes were exactly the same as those of the chip assay. In addition, the in situ hybridization technique was also used to detect the NNAT gene In the retina and the cellular and subcellular distribution of the expressed product.In addition, the search of the chromosomal location of 106 differentially expressed genes revealed that one of them was located in the known retinal cone or cone-rod dystrophy loci Point and with the disease