IMMUNOLOGIAL APPROACHES TO OUTER SEGMENT DISASSEMBLY

外节拆卸的免疫学方法

• 批准号: 3263880
• 负责人: Dennis Michael Defoe
• 金额: $ 6.93万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-30 至 1990-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3263880
• 关键词: Anura; Xenopus; cell adhesion; cell cell interaction; enzyme linked immunosorbent assay; fresh water environment; gel electrophoresis; histochemistry /cytochemistry; immunochemistry; laboratory mouse; 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.

本研究旨在更准确地了解 杆外节（ROS）盘的脱落及其吞噬作用 视网膜色素上皮（RPE）。 加上新碟的更新， 这个过程构成了感光膜的周转，即 维持正常视觉功能所必需的。 脱落吞噬作用是 多步骤过程，可能由多个监管机构协调 阶段。 这些过程包括视细胞外层的正常粘附 RPE 节段，以及有序的椎间盘脱离、吞噬细胞摄取 和退化。 尽管两种细胞类型之间存在协作， 每个人所扮演的具体角色仍然存在疑问。 使用一个过程 分离上皮和视网膜并重建其功能 体外相互作用，我们建议开始分析各个阶段 脱落-吞噬作用。 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解决这个问题 为了识别和表征具有功能的大分子 机械地。 针对上皮细胞和 兔子的光感受器表面会凸起，经过适当的处理后 选择程序，用作视网膜附着和识别的探针 脱落的 ROS 光盘。 此外，还将产生单克隆抗体 RPE 细胞积极参与椎间盘摄取以识别抗原 它在吞噬作用的摄取阶段发挥作用。 这些 免疫球蛋白分子将依次用作试剂 所涉及的细胞成分的生化分离。 希望与 手中有这样的免疫化学探针库，并且具有更好的 细胞相互作用本质的描述，可以在以下方面取得进展 了解ROS反汇编的过程。