VOLUME REGULATION IN CORNEAL ENDOTHELIUM

角膜内皮的体积调节

• 批准号: 6164684
• 负责人: SANGLY P SRINIVAS
• 金额: $ 4.43万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6164684
• 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; 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

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以溶质 - 溶剂耦合和信号通路来支持角膜内皮流体转运的离子传输机制。该项目将分三个步骤进行。在第一步中，将通过以下细胞体积和细胞内离子活性的变化来研究受急性动态扰动的内皮细胞中细胞体积调节的机制。细胞体积的变化将使用光散射，染料稀释和动态荧光猝灭的原理进行测量。眼睛稀释技术的指标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，将确定通过涉及第二使者和蛋白激酶的信号通路调节细胞体积调节。最后，将采用细胞信号传导的操纵，以诱导细胞体积调节的深刻变化来研究其在完整角膜中内皮中对液体转运功能的影响。这种三步方法源于以下假设：驱动矢量流体转运的离子转运活性和溶质 - 溶剂耦合与短期细胞体积调节过程中所涉及的液相同。这项研究的结果对阐明流体运输的机制及其对长期合同晶状体磨损，衰老，糖尿病和眼内外科手术的调节具有直接影响。