VOLUME REGULATION IN CORNEAL ENDOTHELIUM

角膜内皮的体积调节

基本信息

批准号：
6518537
负责人：
SANGLY P SRINIVAS
金额：
$ 20.15万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-09-30 至 2004-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6518537
关键词：
animal tissue biological fluid transport biological signal transduction cell morphology cell osmotic pressure corneal endothelium fluorescent dye /probe green fluorescent proteins intraocular aqueous flow intraocular pressure ion transport laboratory rabbit light scattering physiologic stressor protein kinase second messengers

项目摘要

The principal focus of this proposal is to investigate the ion transport mechanisms that support fluid transport by the corneal endothelium in terms of solute-solvent coupling and signaling pathways. The project will be carried out in three steps. In the first step, the mechanisms of cell volume regulation in endothelial cells subjected to acute anisosmotic perturbations will be investigated by following changes in cell volume and intracellular ionic activities. The changes in cell volume will be measured using the principles of light scattering, dye dilution and dynamic fluorescence quenching. The indicators for eye dilution technique will include transiently expressed green fluorescent protein or polar fluorescent dyes. In the second step, the modulation of cell volume regulation by signaling pathways involving second messengers and protein kinases will be identified. Finally, the maneuvers of cell signaling that induce profound changes in cell volume regulation will be employed to investigate their influence on fluid transport function by the endothelium in intact corneas. This three-step approach stems from the hypothesis that the ion transport activity and solute-solvent coupling that drive the vectorial fluid transport are mostly identical to those involved during short-term cell volume regulation. The results from this study have directly implications towards the elucidation of the mechanisms underlying fluid transport and their modulation upon long-term contract lens wear, aging, diabetes and intra-ocular surgical maneuvers.

该提案的主要重点是研究在溶质-溶剂耦合和信号传导途径方面支持角膜内皮流体运输的离子运输机制。该项目将分三步实施。第一步，将通过跟踪细胞体积和细胞内离子活性的变化来研究遭受急性不等渗扰动的内皮细胞的细胞体积调节机制。利用光散射、染料稀释和动态荧光猝灭的原理来测量细胞体积的变化。眼睛稀释技术的指示剂将包括瞬时表达的绿色荧光蛋白或极性荧光染料。在第二步中，将确定涉及第二信使和蛋白激酶的信号通路对细胞体积调节的调节。最后，将利用细胞信号传导引起细胞体积调节的深刻变化来研究它们对完整角膜内皮的液体运输功能的影响。这种三步方法源于这样的假设：驱动矢量流体传输的离子传输活性和溶质-溶剂耦合与短期细胞体积调节过程中涉及的大部分相同。这项研究的结果对阐明液体运输的机制及其对长期隐形眼镜佩戴、衰老、糖尿病和眼内手术操作的调节具有直接影响。