GENES IN NORMAL AND AMD RETINAS

这项研究计划的主要目标是开始识别和 描述与灵长类动物中央凹相关的基因，并阐明 年龄相关性黄斑变性（AMD）的分子表现。amd是 60岁以上的人中法律的失明的最常见原因 在发达国家，约有10%的老年人 人口与AMD相关的病理变化在以下人群中最普遍： 视网膜黄斑以及脉络膜和视网膜色素上皮中 覆盖这个区域。在黄斑的中心有一个专门的， 无血管区域-中央凹-包含最高密度的锥体 视网膜中的感光细胞。这些中央凹锥在形态上 与中心凹外视锥不同，对于精细的视力至关重要， 似乎更容易在某些人类疾病中退化， 包括AMD。这种对疾病的偏好的基础并不是 了解，并且对有助于基因的基因知之甚少。 中央凹锥体的特殊结构和功能。因此，最初的 本申请的重点将是识别和表征灵长类动物 中心凹视锥特异性和相关cDNA。中央凹特有的转录本 将通过将它们与来自于细胞的转录物进行比较来鉴定视锥细胞。 周边视网膜，使用mRNA差异显示。中心凹特异性 以这种方式鉴定的扩增子将用作探针以分离 来自中央凹富集的cDNA文库的相应全长cDNA。的 人视网膜中央凹cDNA克隆的分子特征和分布 将通过DNA序列分析，原位杂交， 染色体定位和免疫细胞化学。一旦特征化， 中央凹克隆将被用作分子工具，以帮助定义年龄相关的 与正常人视网膜和视网膜的疾病相关变化相比， 从患有AMD的捐赠者的眼睛中获得。其他研究将 目的是鉴定人视网膜特异性cDNA， 作为AMD的结果差异表达。的转录物 特异性地表达于来自被诊断患有 与来自未受影响供体的年龄匹配的视网膜相比， 使用mRNA差异显示鉴定。RNA片段特异性 将使用相同的方法表征在任一组中表达的 概述用于表征中央凹克隆。我们预计， 研究将增加我们对分子结构的整体理解 尤其是中央凹锥体光感受器，并提供 老年性黄斑变性的病因学研究进展

The primary goal of this research program is to begin to identify and characterize genes associated with the primate fovea and to elucidate the molecular manifestations of age-related macular degeneration (AMD). AMD is the most common cause of legal blindness in persons over 60 years of age in developed countries, affecting approximately 10% of the geriatric population. Pathological changes associated with AMD are most prevalent in the retinal macula and in the choroid and retinal pigment epithelium overlying this region. At the center of the macula there is a specialized, avascular region - the fovea - that contains the highest density of cone photoreceptor cells in the retina. These foveal cones are morphologically distinct from extrafoveal cones, are crucial for fine visual acuity and seem to be more susceptible to.degeneration in certain human diseases, including AMD. The bases for this predilection to disease are not understood, and little is known about the genes that contribute to the foveal cones' specialized structure and function. As such, the initial focus of this application will be to identify and characterize primate foveal cone-specific and -associated cDNAs. Transcripts unique to foveal cones will be identified by comparing them to transcripts derived from the peripheral retina, using mRNA differential display. Fovea-specific amplicons identified in this manner will be used as probes to isolate corresponding full-length cDNAs from fovea-enriched cDNA libraries. The molecular characteristics and distribution of the human foveal cDNA clones will be determined by DNA sequence analyses, in situ hybridization, chromosomal mapping and immunocytochemistry. Once characterized, the foveal clones will be used as molecular tools to help define age-related versus disease-related changes in normal human retinas and retinas obtained from eyes of donors afflicted with AMD. Additional studies will be directed toward identifying human retina-specific cDNAs that are differentially expressed as a consequence of AMD. Transcripts that are expressed specifically in retinas from donors with a diagnosed history of AMD, as compared to age-matched retinas from unaffected donors, will be identified using mRNA differential display. RNA fragments specifically expressed in either group will be characterized using the same approach outlined for characterizing foveal clones. We anticipate that these studies will add to our overall understanding of the molecular structure of the fovea, especially the foveal cone photoreceptors, and provide additional insight into the etiology of age-related macular degeneration.