FLUOROPHOTOMETRY OF ION TRANSPORT IN CILIARY BODY

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 房水产生的药理学操作。这个幽默是 由覆盖睫状体的双层上皮产生。 两者，细胞之间的细胞旁过滤(由血液驱动 水相静水压差)和流体的主动运动 这些细胞可能会对这一流动做出贡献。细胞的主动运输， 这已被证明是主要的过程，但人们对此知之甚少。在……里面 在很大程度上，这种有限的知识是由于遇到的困难 在将经典的电学和生理学方法应用于 具有复杂结构的组织中离子和流体传输的研究 解剖学和复杂的细胞排列。这项提议旨在应用小说 用于细胞内离子跟踪的技术(基于细胞可捕获性， 表现出离子敏感光谱的荧光染料)，以产生彻底的 1)存在于两者中的单个转位系统的特征 上皮细胞层；2)上皮细胞层之间的沟通的性质 3)这些活性的药理调节。 专门的设备和解剖技术允许细胞内 单组(或小组)H~+、Na~+、K~+、Cl~-和Ca~(2+)的测定 将使用完整睫状体条带中每种类型的细胞。条带 将安装在灌注室中，允许控制细胞外 组织两侧的离子环境和组织中的变化 由细胞外变化引起的细胞内浓度或 将使用药理效应器来识别和表征每一个 个人运输机。新兴的知识将被应用于生成 房水分泌过程的合理模型。这些型号和 收集到的关于个体药理调节的信息 通过降压剂的运输功能，将提高我们的能力 青光眼患者或个人的房水分泌 表现出高眼压的。

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 humor production. This humor is generated by the dual layered epithelium that covers the ciliary body. Both, paracellular filtration between the cells (driven by the blood to aqueous hydrostatic pressure difference) and active movement of fluid by the cells could contribute to the flow. 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 novel techniques for intracellular ion tracking (based on cell entrapable, fluorescent dyes exhibiting ion sensitive spectra) to generate a thorough characterization of 1) the individual translocation systems present in both epithelial cell layers; 2) the nature of the communication between the layers; and 3) the pharmacological regulation of these activities. Specialized equipment and dissection techniques allowing intracellular measurements of H+, Na+, K+, Cl- and Ca2+ in single (or small groups of) cells of each type in intact ciliary body strips will be used. The strips will be mounted in perfusion chambers allowing control of the extracellular ionic environment in each side of the tissue and the changes in intracellular concentrations induced by extracellular changes or by pharmacological effectors will be used to identify, and characterize each individual transporter. The emerging knowledge will be applied to generate plausible models for the aqueous humor 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.