BIOCHEMICAL RESPONSES OF HUMAN CILIARY EPITHELIAL CELLS

人睫状上皮细胞的生化反应

• 批准号: 3259434
• 负责人: MIGUEL COCA-PRADOS
• 金额: $ 15.46万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3259434
• 关键词: active transport; adenosinetriphosphatase; adenylate cyclase; biochemistry; calcium; cell sorting; chromatography; cyclic AMP; density gradient ultracentrifugation; epithelium; fluorescence; fluorescence microscopy; gel electrophoresis; glaucoma; human tissue; hybridomas; immunofluorescence technique; immunologic techniques; inositol phosphates; ion transport; monoclonal antibody; phosphorylation; protein kinase; protein kinase C; radioassay; secretion; sympathetic nervous system; tissue /cell culture; uvea ciliary body

Knowledge of aqueous humor secretion and regulation of aqueous production in the human eye is relevant to the understanding and treatment of glaucoma. Aqueous humor is secreted by the ciliary epithelium and its composition is largely dependent on the mechanism of active transport of plasma proteins an electrolites in these cells. The rate of formation of aqueous humor is modulated by the sympathetic (adrenergic) system (Neufeld, A.H., 1984). The development of a tissue culture system for the ciliary epithelial (CE) cells, namely the nonpigmented (NPE) and pigmented (PE) epithelia is important because it permits the in vitro study of the cellular and molecular functions of these cells which are very difficult to carry out in vivo. The aim of this proposal is to develop antibodies to both membrane-bound surface proteins and internal proteins which are specific markers for NPE and PE cells using the hybridoma technology. These antibodies will serve as probes to investigate the differentiated properties of these cells in culture. In addition, it will allow for the isolation (i.e. fluorescence activated cell sorter) of NPE and PE cells based on their potential immunological differences. These antibodies will be used to facilitate the understanding of the secretory function and transport of NPE and PE cells. Secretory function will be studied by developing antibodies to a 76KDa protein secreted by CE in culture. Ion transport will be studied by immunobloting analysis of (Na+, K+)-ATPase activity by antibodies to the alpha subunit of this enzyme. Sodium pump transport activity will assessed by ouabain-sensitive 86Rb+ uptake. In addition, the mechanism of transduction by external signals into intracellular events (i.e., protein phosphorylation) will be studied.

了解水性幽默分泌和水的调节 人眼的生产与理解和 青光眼治疗。 水性幽默被睫状分泌 上皮及其组成在很大程度上取决于 血浆蛋白主动转运的机制 在这些细胞中。 幽默的形成速率是 通过交感神经（肾上腺素能）系统（Neufeld，A.H。， 1984）。 睫状组织组织培养系统的发展 上皮（CE）细胞，即未浸润（NPE）和 色素（PE）上皮很重要，因为它允许IN 体外研究这些细胞的细胞和分子功能 在体内执行非常困难。 该提议的目的是开发两者的抗体 膜结合的表面蛋白和内部蛋白 使用杂交瘤的NPE和PE细胞的特定标记 技术。 这些抗体将作为调查的探针 这些细胞在培养中的分化特性。 此外， 它将允许分离（即荧光激活的细胞 NPE和PE单元的分子基于其电​​势 免疫学差异。 这些抗体将用于促进理解 NPE和PE细胞的分泌功能和运输。 分泌功能将通过开发抗体来研究 76KDA蛋白在培养中分泌。 离子运输将是 通过（Na+，K+）ATPase活性的免疫印迹分析研究 通过该酶的α亚基的抗体。 钠泵 运输活动将由瓦巴因敏感的86RB+评估 吸收。 另外，外部信号转导的机制 进入细胞内事件（即蛋白质磷酸化）将是 研究。