MODEL OF HEREDITARY BLINDNESS--PROTEIN ANALYSES

国际和平研究所研究计划的长期目标是确定和 了解视网膜光感受器细胞的生化原因 功能障碍。这样的信息将具有深远的医学意义 并可能提供对生物医学变化的洞察，这些变化是导致心脏功能丧失的原因 人类视网膜疾病中的视网膜功能。《红楼梦》的视网膜分析 具有损害光感受器细胞的基因突变的动物 功能和可行性为实现这一目标提供了一种权宜之计 进球。拟议的研究将侧重于确定和理解 视网膜色素变性与光感受器功能障碍相关的蛋白质变化 (视网膜变性)雏鸟。对RD雏鸡视网膜的研究提供了一个 调查潜在的生化变化的独特机会 视细胞功能完全丧失所致的感光细胞功能障碍 这些动物的视网膜是在没有大体发育的情况下发生的 异常和先于细胞退化。 RD突变的主要生化位点(S)尚不清楚。 蛋白质双向凝胶电泳法的初步分析 从纯合子Rd[Rd/Rd]、杂合载体[+/Rd]和 正常的[+/+]视网膜在发育期间和出现之前 已经发现了两组蛋白质，它们的表达是 受到突变的影响。拟议的实验将提供 关于这些蛋白质的生化特性的信息，它们的 组织特异性及其在组织内的细胞和亚细胞位置 产生它们的组织。氨基酸分析、肽图和N- 这些蛋白质的末端或内部多肽氨基酸序列将 用来确定蛋白质之间的同源性程度。 针对这些蛋白制备的单特异性抗血清将用于 确定产生蛋白质的细胞及其细胞内 地点。这些数据和氨基酸序列数据将提供 关于这些蛋白质的细胞功能的重要线索。 总而言之，这些结果将提高我们对生物化学的理解 RD雏鸡视网膜光感受器功能障碍的改变 可提供对导致营养物质损失的一般生化过程的洞察 视网膜疾病中的光感受器功能。

The long-term objective of the PI's research program is to identify and understand the biochemical cause or causes of retinal photoreceptor cell dysfunction. Such information would be of profound medical significance and may provide insight into the biomedical changes which underlie loss of retinal function in diseases of the human retina. Analyses of retinas of animals possessing genetic mutations which compromise photoreceptor cell function and viability provide an expedient means for accomplishing this goal. The proposed studies will focus on identifying and understanding protein changes associated with photoreceptor dysfunction in retinas of rd (retinal degeneration) chicks. Studies of the rd chick retina provide a unique opportunity to investigate biochemical changes underlying photoreceptor cell dysfunction because total loss visual cell function in the retinas of these animals occurs in the absence of gross developmental abnormalities and precedes cellular degeneration. The primary biochemical site(s) for the rd mutation is not known. Preliminary two-dimensional gel electrophoretic analyses of proteins extracted from homozygous rd [rd/rd], heterozygous carrier [+/rd] and normal [+/+] retina during development and prior to the appearance of degeneration have revealed two groups of proteins whose expression is affected by the mutation. The proposed experiments will provide information concerning the biochemical identities of these proteins, their tissue-specificity and their cellular and subcellular locations within the tissues which produce them. Amino acid analyses, peptide maps and N- terminal or internal peptide amino acid sequences of these proteins will be used to determine the degree of homology between the proteins. Monospecific antisera raised against these proteins will be used to identify the cells which produce the proteins and their intracellular location. These data along with the amino acid sequence data will provide important clues regarding the cellular functions of these proteins. Together, these results will improve our understanding the biochemical changes underlying photoreceptor dysfunction in the rd chick retina and may provide insight into general biochemical processes which cause loss of photoreceptor function in retinal disease.