BIOCHEMICAL RESPONSES OF HUMAN CILIARY EPITHELIAL CELLS

了解房水的分泌和调节 在人眼中的产生与理解有关， 青光眼的治疗 眼房水是由睫状体分泌的 上皮及其组成在很大程度上取决于 血浆蛋白和电解质主动转运机制 在这些细胞中。 房水的形成率是 由交感（肾上腺素能）系统调节（Neufeld，A.H.， 1984年）。 睫状体组织培养体系的建立 上皮（CE）细胞，即无色素（NPE）， 色素上皮（PE）是重要的，因为它允许在 这些细胞的细胞和分子功能的体外研究 这是很难在体内进行的。 这项提案的目的是开发针对这两种病毒的抗体。 膜结合的表面蛋白和内部蛋白， 使用杂交瘤的NPE和PE细胞的特异性标志物 技术. 这些抗体将作为探针， 这些细胞在培养中的分化特性。 此外，本发明还提供了一种方法， 它将允许分离（即荧光激活的细胞 分类器）的NPE和PE细胞的基础上，他们的潜力 免疫学差异 这些抗体将用于促进理解 NPE和PE细胞的分泌功能和转运。 分泌功能将通过开发抗体来研究， CE在培养物中分泌76KDa蛋白。 离子传输将是 通过免疫印迹分析（Na+，K+）-ATP酶活性研究 通过这种酶的α亚基的抗体。 钠泵 通过哇巴因敏感的86Rb+评估转运活性 摄取。 此外，通过外部信号转导的机制 转化为细胞内事件（即，蛋白质磷酸化）将是 研究了

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.