ION FLUOROPHOTOMETRY OF CILIARY BODY EPITHELIA

稳态眼内压是以下因素之间平衡的结果： 流体产生和流出。 眼内压（IOP）升高，a 可能对视力造成破坏性影响的疾病， 退化，当流出设施减少时发生。 然而，在这方面， 缓解IOP升高的临床干预主要依赖于 药理学操纵水的人类生产。 这种幽默是 由合胞排列的双层上皮细胞产生， 睫状体。 细胞主动运输，已被证明， 作为主要的过程，人们对此知之甚少。 在很大程度上，这一限制 知识是由于在应用中遇到的困难， 经典的电学和生理学方法来研究离子和 在具有复杂解剖结构和复杂组织的组织中的流体输送 细胞排列 该提案旨在应用非侵入性光学 用于细胞内离子追踪的技术（基于可捕获的细胞， 显示离子敏感光谱的荧光染料）以产生彻底的 表征1）钙动员现象及其调节， CBE各层及整个组织中的离子现象 控制细胞内pH和碳酸氢盐通量; 3）功能性 两种CBE之间通讯的性质和生化基础 层;和这些活动的药理学调节。 专业的设备和解剖技术， 测量单个（或小组）细胞中的H+和Ca 2+， 将使用每种类型或同时在两种细胞类型中使用。 电生理和免疫组织学研究将用于 补充基于荧光的方法。 新兴的知识将是 应用于生成合理的模型，为水的人类分泌 过程 这些模型和收集的药理学信息 通过膨胀剂调节个体运输功能将 提高我们控制青光眼房水分泌能力 表现出IOP升高的患者或个体。

The steady state intraocular pressure is the result of a balance between fluid production and outflow. Elevated intraocular pressure (IOP), a condition which could have devastating effects for vision due to tissue degeneration, occurs when the outflow facility is reduced. However, clinical intervention to relieve elevated IOP relies mostly in pharmacological manipulation of aqueous human production. This humor is generated by the syncytially arranged dual layered epithelium that covers the ciliary body. The cellular active transport, which has been shown to be the primary process, is poorly understood. In great part, this limited knowledge is due to the difficulties encountered in the application of classical electrical and physiological approaches to the study of ion and fluid transport in a tissue endowed with an intricate anatomy and a complex cell arrangement. This proposal aims to apply noninvasive optical techniques for intracellular ion tracking (based on cell entrappable, fluorescent dyes exhibiting ion sensitive spectra) to generate a thorough characterization of 1) calcium mobilization phenomena and its regulation in each layer of the CBE and in the whole tissue; 2) the ionic phenomena controlling intracellular pH and bicarbonate flux; 3) the functional properties and biochemical basis of the communication between the two CBE layers; and the pharmacological regulation of these activities. Specialized equipment and dissection techniques allowing intracellular measurements of H+ and Ca2+ in either single (or small groups of) cells of each type or, in both cell types simultaneously, will be used. Electrophysiological and immunohistological investigation will be used to complement the fluorescence-based methods. The emerging knowledge will be applied to generate plausible models for the aqueous human secretory process. These models and the information gathered on the pharmacological modulation of individual transport functions by hypotensive agents will improve our capacity to manipulate aqueous humor secretion in glaucoma patients or individuals exhibiting an elevated IOP.