ION FLUOROPHOTOMETRY OF CILIARY BODY EPITHELIA

稳定状态下的眼压是下列因素平衡的结果 流体的产生和流出。眼压升高，a 由于组织的原因可能对视力造成毁灭性影响的情况 当外流设施减少时，就会发生退化。然而， 临床干预缓解高眼压主要依赖于 人类生产含水产品的药理学操作。这个幽默是 由合胞排列的双层上皮产生，覆盖 睫状体。细胞主动运输，这已经被证明是 作为主要过程，人们对此知之甚少。在很大程度上，这是有限的 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研究离子和 具有复杂解剖结构和复杂结构的组织中的流体传输 单元格排列。该提案旨在应用非侵入性光学技术 用于细胞内离子跟踪的技术(基于细胞可携带， 表现出离子敏感光谱的荧光染料)，以产生彻底的 1)钙动员现象的特征及其调控 CBE的每一层和整个组织；2)离子现象 控制细胞内pH和碳酸氢盐通量；3)功能 两种CBE之间通讯的性质和生化基础 以及对这些活动的药理调节。 专门的设备和解剖技术允许细胞内 单细胞(或一小群细胞)H+和Ca~(2+)的测定 将使用每种类型或同时在两种单元类型中使用。 电生理和免疫组织学检查将用于 补充了基于荧光的方法。新兴的知识将是 应用于为人类分泌物生成可信的模型 进程。这些模型和收集的关于药理学的信息 降压药对个体转运功能的调节将 提高青光眼患者控制房水分泌的能力 眼压升高的患者或个人。

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.