IMMUNOLOGIAL APPROACHES TO OUTER SEGMENT DISASSEMBLY

外节拆卸的免疫学方法

• 批准号: 3263875
• 负责人: Dennis Michael Defoe
• 金额: $ 7.72万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-30 至 1989-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3263875
• 关键词: cell adhesion; cell cell interaction; enzyme linked immunosorbent assay; fresh water environment; gel electrophoresis; histochemistry /cytochemistry; immunochemistry; ligands; monoclonal antibody; phagocytosis; retina; retinal pigment epithelium; rod cell; tissue /cell culture; visual photoreceptor

This research is directed toward a more precise understanding of the shedding of rod outer segment (ROS) discs and their phagocytosis by the retinal pigment epithelium (RPE). Together with the renewal of new discs, this process constitutes the turnover of photoreceptive membrane which is necessary to maintain proper visual function. Shedding-phagocytosis is a multistep process which is probably coordinated by regulation at several stages. These processes include the normal adhesion of visual cell outer segments to the RPE, as well as orderly disc detachment, phagocytic uptake and degradation. Although a collaboration between both cell types, the specific roles played by each are still in doubt. Using a procedure for separating epithelium and retina and reconstituting their functional interactions in vitro, we propose to begin analyzing various stages in shedding-phagocytosis. To complement this approach, highly sensitive and selective immunochemical techniques will be applied to this problem in order to identify and characterize macromolecules which function mechanistically. Heteroantisera directed towards epithelial and photoreceptor surfaces will be raised in rabbits and, after appropriate selection procedures, used as probes of retinal attachment and recognition of shed ROS discs. In addition, monoclonal antibodies will be generated to RPE cells actively engaged in disc uptake in order to identify antigens which function in the ingestion phase of phagocytosis. These immunoglobulin molecules will in turn be used as reagents for the biochemical isolation of cell constituents involved. It is hoped that with an arsenal of such immunochemical probes in hand and with a better description of the nature of cellular interactions, progress can be made in understanding the process of ROS disassembly.

这项研究是针对对 杆外段（ROS）椎间盘的脱落及其吞噬作用 视网膜色素上皮（RPE）。 与新光盘的更新一起 此过程构成感光膜的营业额 保持适当的视觉功能所必需的。 脱落 - 细胞增多症是 多步过程，可能通过几个调节协调 阶段。 这些过程包括视觉细胞外部的正常粘附 RPE的细分市场以及有序的椎间盘脱离，吞噬摄取 和退化。 尽管两种单元类型之间的合作，但 每个人扮演的具体角色仍然令人怀疑。 使用一个程序 分离上皮和视网膜并重新建立其功能 在体外相互作用，我们建议开始分析各个阶段 脱落 - 细胞增多症。 为了补充这种方法，高度敏感和 选择性免疫化学技术将应用于此问题 为了识别和表征大分子的功能 机械上。 杂质，针对上皮和 光感受器表面将在兔子中抬高，并在适当的 选择程序，用作视网膜依恋和识别的探针 Shed Ros Discs。 此外，将产生单克隆抗体 RPE细胞积极参与椎间盘吸收以识别抗原 哪个在吞噬作用的摄入阶段。 这些 免疫球蛋白分子反过来又将用作试剂 涉及细胞成分的生化分离。 希望与 此类免疫化学探针的武器库，并且更好 细胞相互作用的性质的描述，可以在 了解ROS拆卸的过程。