REGULATION OF PHI AND FLUID FLUX IN CORNEAL ENDOTHELIUM

清晰的视力需要光学透明的角膜。透明度是 依赖于角膜水化，这是由上皮细胞控制的 细胞层称为角膜内皮。内皮控制 通过从角膜吻合口向房水中分泌液体进行水化作用 通过离子耦合流体传输的体液。膜与细胞 内皮细胞转运的调节机制不是 明白了。PI的目标是开发一种内皮细胞液体模型 转运包括：特定的膜转运蛋白 涉及到的，它们的运输极性，以及运输如何 受监管的。一个长期目标是了解疾病是如何发生的(例如，Fuchs的 血管内皮细胞营养不良，糖尿病)改变血管内皮功能。 可以想象，医学疗法可以被开发来延迟或取代 角膜内皮病或角膜移植的必要性 精神创伤。由于内皮细胞的运输需要HC03和氯，我们将 检查内皮细胞调节其功能的机制 胞内pH(Phi)、控制胞内(C1)和c1转运 速率，使用离子敏感的荧光探针测定pH、Na+、C1和 Ca2+。转运体的极性由以下因素决定：a)生理 对生长在可渗透滤光片上的细胞的测量，b)光和电子 显微镜免疫细胞化学和c)免疫印迹 顶端/基侧膜组分。碳酸酐酶抑制剂 血管内皮细胞液体运输缓慢。因此，我们将研究这些角色 胞质结合和膜结合碳酸酐酶在HC03中的变化 二氧化碳。将通过测量HC03的影响来检验这一点 非搅拌层pH上的转运和碳酸酐酶阻滞剂 和液体运输。由于流体传输依赖于HC03，因此 PHI在确定流体运输中的作用将被检验。这些 研究将为未来医学的发展提供基础 恢复伴随的视力丧失的治疗方法 内皮疾病或创伤。

Clear vision requires an optically transparent cornea. Transparency is dependent on corneal hydration, which is controlled by an epithelial cell layer called the corneal endothelium. The endothelium controls hydration by secreting fluid from the corneal stoma to the aqueous humor by ion coupled fluid transport. The membrane and cellular regulatory mechanisms of transport in the endothelium are ot understood. The PI's goal is to develop a model for endothelial fluid transport that includes: the specific membrane transport proteins involved, their polarity of transport, and how transport can be regulated. A long-term goal is to udnerstand how disease, (e.g. fuchs' Endothelial Dystrophy, Diabetes) alters endothelial function. Conceivable, medical therapies can be developed to delay or supplant the need for corneal transplantation in cases of endothelial disease or trauma. Since endothelial transport requires HC03 and Cl, we will examine the mechanisms by which endothelial cells regulate their intracellular pH (pHi), control intracellular (C1) and C1 transport rates, using ion sensitive fluorescent probes for pH, Na+, C1 and Ca2+. Transporter polarity will be determine by: a) physiological measurements on cells grown on permeable filters, b) light and elctron microscope immunocytochemistry and c) western blotting of apical/basolateral membrane fractions. Carbonic anhydrase inhibitors slow endothelial fluid transport. Therefore we will examine the roles of cytoplasmic and membrane bound carbonic anhydrase in HC03 to C02. This will be examined by measuring the effects of HC03 transport and carbonic anhydrase blockers on the unstirred layer pH and fluid transport. Because fluid transport is HC03 dependent, the role of pHi in determining fluid transport will be examined. These studies will provide the basis for future development of medical therapies for restoring the loss of vision that can accompany endothelial disease or trauma.