Differential Gene Expression in Ciliary Epithelium

描述(申请人提供)：降低初中生眼压 开角型青光眼是青光眼最常见的形式，是主要的目标 在治疗中防止这种眼病的进展。我们的 长期目标是在细胞和分子水平上理解 参与眼压调节的机制。一大群人 在很大程度上，来自分子生物学研究的证据表明 支持人类睫状体上皮是神经内分泌的，并表达一种 功能性多肽能系统。许多已鉴定的多肽具有血管活性。 它们参与心血管系统中血压的调节。 系统。睫状体上皮细胞内肽能系统的存在 为它参与眼睛的调节提供了提示性的证据 通过自分泌/旁分泌机制在“局部”水平施加压力。 在本申请中，我们提出了一项为期5年的研究，以检验以下假设 眼压是睫状体上皮的神经内分泌功能， 通过携带的调节肽和激素的表达而介导 昼夜节律是由一种非视觉感光机制实现的。 该计划包括三个主要目标。目标1，生化特性，由 神经内分泌调节肽的高效液相色谱和质谱学研究 在纤毛上皮细胞中表达的高血压/降压活动， 包括神经肽Y和利钠肽-C。研究它们的信号 通过测量秒激活它们的同源受体的途径 信使包括cAMP、cGMP和钙离子。目标2，检查昼夜节律 神经肽在实验动物和培养细胞模型中的变化 睫状体上皮进入明/暗周期。要确定 构成昼夜节律的内源“昼夜节律振荡器”的组成部分 睫状体上皮细胞的基因表达，包括褪黑素节律的产生 酶和时钟基因。目标3，确定该病毒的分子性质 通过聚焦，睫状体上皮中的非视觉光转导级联 视紫红质和其他视蛋白样分子的表达，以及 视紫红质激活和失活的成分， 包括视紫红质激酶、恢复素和芦荟素。要确定任何 完整的和培养的睫状体上皮细胞的光敏反应，通过 研究了cGMP门控通道在明暗条件下的变化。

DESCRIPTION (provided by applicant): Lowering intraocular pressure in Primary Open Angle Glaucoma, the most prevalent form of glaucoma, is a major objective in the treatment to prevent the progression of this ocular disease. Our long-term goal is the understanding at the cell and molecular level of the mechanisms involved in the regulation of intraocular pressure. A great body of evidence emerging, for the most part, from studies on molecular biology has supported that the human ciliary epithelium is neuroendocrine and expresses a functional peptidergic system. Many of the peptides identified are vasoactive and they are involved in the regulation of blood pressure in the cardiovascular system. The existence of a peptidergic system in the ciliary epithelium provides suggestive evidence for its involvement in the regulation of eye pressure at a "local" level by autocrine/paracrine mechanisms. In this application, we propose a 5-year study to examine the hypothesis that intraocular pressure is a neuroendocrine function of the ciliary epithelium, mediated by the expression of regulatory peptides and hormones entrained in a circadian rhythm by a non-visual light-sensing mechanism. The plan comprises three main aims. Aim 1, biochemical characterization, by HPLC and mass spectrometry, of neuroendocnne regulatory peptides with hypertensive/hypotensive activities expressed in the ciliary epithelium, including neuropeptide Y and natriuretic peptide-C. To study their signaling pathway upon activation of their cognate receptors by measuring second messengers including cAMP, cGMP and Ca2+. Aim 2, to examine the circadian variations of neuropeptides in experimental animals and cultured cell models of the ciliary epithelium entrained to light/dark cycles. To identify the components of the endogenous "circadian oscillator" that underlie the circadian gene expression in the ciliary epithelium, including melatonin-rhythm-generating enzymes and clock genes. Aim 3, to establish the molecular nature of the non-visual phototransduction cascade in the ciliary epithelium, by focusing on the expression of rhodopsin and other opsin-like molecules, and components of the activation and deactivation of rhodopsin, including rhodopsin kinase, recoverin and arrestin. To determine any light-sensing responses in intact and cultured ciliary epithelial cells, by studying the cGMP-gated channel under light and dark conditions.