SIGNAL TRANSDUCTION BY G-PROTEINS IN THE RETINAL PE

本提案中提出的研究目标是获得 了解视网膜色素的协调生理学 上皮（RPE）和神经视网膜。 在众多RPE功能中， 对于神经视网膜的生存能力至关重要的是对 光感受器膜脱落的包。 拟议的研究重点是 RPE吞噬作用的信号转导过程，以及 基于鸟嘌呤介导的信号传导事件 核苷酸结合蛋白（G蛋白）可能在细胞凋亡中发挥特定作用。 RPE和光感受器之间的相互作用。 具体目标是： 1)为了鉴定G蛋白和G蛋白偶联受体， 通过克隆和测序相应的cDNA， 2)建立 G蛋白和G蛋白的组织特异性和亚细胞定位 蛋白偶联受体 3)为了表征RPE特异性的功能， 蛋白质，并探讨其在信号转导中的潜在作用 与吞噬作用有关的过程。 实验策略将是 基于分子生物学的技术，并将利用高 存在于G蛋白之间和G蛋白之间的同源性程度。 受体。 北方分析，原位杂交，和 免疫细胞化学将用于确定组织特异性， 亚细胞定位。 配体结合和吞噬作用的测定将 用于确定RPE特异性蛋白的功能。 本研究 将有助于信号传导电位的定义和细胞 分子水平上的RPE生物学。 长期目标是 视网膜变性的病因是什么？ RPE和视网膜之间的关系中断。

The objective of the research set out in this proposal is to gain an understanding of the coordinate physiology of the retinal pigment epithelium (RPE) and the neural retina. Among the many RPE functions critical for the viability of the neural retina is the phagocytosis of packets of shed photoreceptor membranes. The proposed research focuses on the signal transduction processes underlying phagocytosis by the RPE, and is based on the hypothesis that signaling events mediated by Guanine nucleotide-binding proteins (G-proteins) may play a specific role in the interactions between the RPE and photoreceptors. The specific aims are: 1) To identify the G-proteins and G-protein-coupled receptors present in the RPE by cloning and sequencing corresponding cDNAs. 2) To establish the tissue specificity and subcellular localization of the G-proteins and G protein-coupled receptors. 3) To characterize the function of RPE specific proteins and explore their potential roles in the signal transduction processes associated with phagocytosis. The experimental strategy will be based in the techniques of molecular biology, and will make use of the high degree of homology that exists among the G-proteins and among the receptors. Northern analysis, in situ hybridization, and immunocytochemistry will be used to determine tissue specificity and subcellular localization. Assays of ligand binding and phagocytosis will be used to determine the function of RPE specific proteins. This research will contribute to the definition of the signaling potential and cell biology of the RPE on a molecular level. The long term goals are to relate this knowledge to the underlying causes of retinal degeneration, in which the relationship between the RPE and retina is interrupted.