mRNA expression by RPE cells in myopia

描述（由申请人提供）：许多证据表明视网膜在很大程度上调节眼睛的生长和屈光不正的发展，但是视网膜控制体内巩膜生长的信号级联的细胞途径和分子成分的身份尚不清楚。我们建议研究单个视网膜色素上皮（RPE）细胞的mRNA表达，以解决眼睛生长控制中局部视网膜-巩膜信号，因为RPE细胞形成感觉视网膜和脉络膜/巩膜之间的中间屏障层，并且根据现有数据，可以局部介导视网膜对屈光发育的影响。我们将在两种成熟的鸡眼生长模型中选择不同眼底区域的RPE细胞，通过对这些细胞mRNA制备的扩增反义rna （aRNA）进行微阵列分析，研究其基因表达。

DESCRIPTION (provided by applicant): Much evidence indicates that the retina largely regulates eye growth and the development of refractive errors, but the identity of both the cellular pathways and the molecular components of the signaling cascade by which the retina controls scleral growth in vivo are poorly understood. We propose to investigate mRNA expression by individual retinal pigment epithelial (RPE) cells to address local retina-to-sclera signaling in eye growth control because RPE cells form the intervening barrier layer between the sensory retina and the choroid/sclera and, based on available data, could locally mediate the retinal influences on refractive development. We shall study gene expression in individual RPE cells selected from different fundus regions in two well-established eye growth models in the chick by applying microarray profiling to amplified antisense-RNA (aRNA) prepared from the mRNA of these cells. Combining an interdisciplinary research team and University of Pennsylvania core facilities, we propose the following inter-related Specific Aims: 1) prepare aRNA from the mRNA of individual RPE cells located at different fundus regions in eye growth models; 2) complete and then utilize high density chick RPE/retinal microarrays to profile aRNA from RPE cells; 3) adapt informatics approaches for analysis of gene expression by individual RPE cells; and 4) validate the cellular gene expression profiles. By identifying mRNA expression patterns according to RPE cell location, we anticipate that the results will further implicate RPE cells in the growth regulatory process and provide novel molecular and spatial information about the local signaling from retina to sclera in refractive development. Further, demonstrating the feasibility and utility of single cell mRNA profiling for refractive research could substantially advance the available techniques and provide the foundation for future cell and molecular studies in the area. We believe this program will lead to improved understanding of the etiology of refractive errors and ultimately to effective approaches to arrest myopia in children.