MODEL OF HEREDITARY BLINDNESS--PROTEIN ANALYSES

PI研究计划的长期目标是确定和 了解视网膜感光细胞的生化原因或原因 功能障碍 这些信息将具有深远的医学意义 并可能提供深入了解生物医学变化的基础损失， 人类视网膜疾病中的视网膜功能。 视网膜分析 具有损害感光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有利的手段 目标. 拟议的研究将侧重于查明和了解 视网膜变性患者视网膜光感受器功能障碍相关的蛋白质变化 （视网膜变性）小鸡。 对rd小鸡视网膜的研究提供了一个 独特的机会来研究潜在的生物化学变化 光感受器细胞功能障碍是因为视觉细胞功能完全丧失， 这些动物的视网膜发生在没有总体发育的情况下， 异常和细胞变性之前。 rd突变的主要生化位点尚不清楚。 蛋白质的初步二维凝胶电泳分析 从纯合rd [rd/rd]、杂合携带者[+/rd]和 正常[+/+]视网膜在发展过程中和之前的外观 退化揭示了两组蛋白质，其表达是 受到突变的影响。 这些实验将提供 关于这些蛋白质的生物化学特性的信息， 组织特异性及其在组织中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肽氨基酸序列将 用于确定蛋白质之间的同源性程度。 针对这些蛋白质产生的单特异性抗血清将用于 鉴定产生蛋白质的细胞及其细胞内 位置. 这些数据沿着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。 总之，这些结果将提高我们对生物化学的理解。 在RD小鸡视网膜中潜在的光感受器功能障碍的变化， 可以提供洞察一般的生化过程，造成损失的 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.