Region-specific Transcriptome Analysis of the Human Retina and RPE/Choroid. Region-specific Transcriptome Analysis of the Human Retina and RPE/Choroid

Proper spatial differentiation of retinal cell types is necessary for normal human vision. Many retinal diseases, such as Best disease and male germ cell associated kinase (MAK)-associated retinitis pigmentosa, preferentially affect distinct topographic regions of the retina. While much is known about the distribution of cell types in the retina, the distribution of molecular components across the posterior pole of the eye has not been well-studied. To investigate regional difference in molecular composition of ocular tissues, we assessed differential gene expression across the temporal, macular, and nasal retina and retinal pigment epithelium (RPE)/choroid of human eyes using RNA-Seq. RNA from temporal, macular, and nasal retina and RPE/choroid from four human donor eyes was extracted, poly-A selected, fragmented, and sequenced as 100 bp read pairs. Digital read files were mapped to the human genome and analyzed for differential expression using the Tuxedo... (for more see dbGaP study page.)

视网膜细胞类型的精准空间分化，是维持人类正常视觉功能的必要前提。诸多视网膜疾病，例如贝斯特病（Best disease）以及雄性生殖细胞相关激酶（male germ cell associated kinase, MAK）相关的视网膜色素变性，均偏好性累及视网膜的特定地形学区域。尽管学界对视网膜内细胞类型的分布已有较多研究，但针对眼球后极部区域内分子组分的分布特征，目前尚缺乏充分的研究。为探究眼部组织分子构成的区域差异，本研究采用RNA测序（RNA-Seq）技术，对人类眼球的颞侧视网膜、黄斑区视网膜、鼻侧视网膜以及视网膜色素上皮（retinal pigment epithelium, RPE）/脉络膜组织进行了差异基因表达分析。研究人员从4例人类供体眼球中提取颞侧视网膜、黄斑区视网膜、鼻侧视网膜以及RPE/脉络膜的总RNA，经polyA富集、片段化处理后，构建为100bp双端测序文库并完成测序。将数字化测序读段文件比对至人类参考基因组，并采用Tuxedo套件进行差异表达基因分析……（详细内容请参见dbGaP研究页面）