Ion Conductances in the Retinal Pigment Epithelium

描述（由申请人提供）：光感受器的健康和完整性严重依赖于其细胞外微环境的组成和体积。这种所谓的视网膜下空间的离子组成和体积的调节是通过离子和水在视网膜色素上皮（RPE）上的运输来完成的，RPE是一种单层细胞，位于光感受器外段和脉络膜血液供应之间。RPE转运是多种离子转运蛋白和通道协同活动的结果，这些蛋白和通道位于其顶端和基底膜上。随着视网膜活动的变化，视网膜细胞释放的化学信号扩散到RPE，并开始调整其运输，以补偿光感受器微环境的变化。这些运输过程或其调控的破坏可能导致视网膜下空间的不利变化，从而导致视网膜疾病。这些运输途径也负责维持RPE细胞的细胞内组成，如果受到干扰，可能会对其他关键的RPE功能产生不利影响，如维生素A的运输和代谢。我们的总体目标是了解钾（K+）通道参与调节视网膜下间隙和RPE细胞质中液体的体积和离子组成的机制。具体目的是：(1)确定RPE中m型K+通道的分子基础；(2)确定Kir7.1以外的内向整流K+ (Kir)通道亚基是否有助于RPE K+电导；(3)了解细胞内pH的生理变化如何调节Kir7.1通道；(4)阐明m型K+通道受体激活和抑制的信号转导途径。这些目标将通过结合分子和电生理技术来研究K+通道的结构、功能和调节。这些研究的结果将更好地了解这些至关重要的转运蛋白如何在RPE中运作以维持健康的光感受器微环境。

DESCRIPTION (provided by applicant): The health and integrity of photoreceptors critically depend on the composition and volume of their extracellular microenvironment. Regulation of the ionic composition and volume of this so-called subretinal space is accomplished by the transport of ions and water across the retinal pigment epithelium (RPE), a monolayer of cells juxtaposed between the photoreceptor outer segments and the choroidal blood supply. RPE transport is the result of the coordinated activity of a diverse group of ion transport proteins and channels residing in its apical and basolateral membranes. With changes in retinal activity, chemical signals released by retinal cells diffuse to the RPE and initiate adjustments in its transport to compensate for alterations in the photoreceptor microenvironment. Disruption of these transport processes or their regulation may cause adverse changes in the subretinal space, contributing to retinal disease. These transport pathways are also responsible for maintaining the intracellular composition of the RPE cell, which, if disturbed, could adversely affect other key RPE functions such as vitamin A transport and metabolism. Our overall goal is to understand the mechanisms by which potassium (K+) channels participate in the regulation of the volume and ionic composition of the fluid in both the subretinal space and the RPE cytoplasm. The specific aims are: (1) To determine the molecular basis for the M-type K+ channel in the RPE; (2) To ascertain whether inwardly rectifying K+ (Kir) channel subunits other than Kir7.1 contribute to the RPE K+ conductance; (3) To understand how the Kir7.1 channel is modulated by physiological changes in intracellular pH; and (4) To elucidate the signal transduction pathway linking receptor activation and inhibition of M-type K+ channels. These aims will be pursued using a combination of molecular and electrophysiological techniques to investigate K+ channel structure, function, and regulation. The outcome of these studies will be a better understanding of how these critically important transport proteins operate in the RPE to maintain a healthy photoreceptor microenvironment.