Differential Gene Expression in Ciliary Epithelium

描述（由申请人提供）：降低原发性眼内压 开角型青光眼是青光眼中最常见的一种， 在治疗中防止这种眼病的发展。我们 长期目标是在细胞和分子水平上理解 调节眼内压的机制。一大批 大多数来自分子生物学研究的证据， 支持人睫状上皮是神经内分泌的，并表达 功能肽能系统。许多已鉴定的肽具有血管活性 它们参与了心血管系统中血压的调节 系统睫状体上皮中存在肽能系统 提供了暗示性的证据，其参与调节眼睛 通过自分泌/旁分泌机制在“局部”水平上产生压力。 在本申请中，我们提出了一项为期5年的研究，以检验假设， 眼内压是睫状上皮的神经内分泌功能， 通过调节肽和激素的表达介导， 昼夜节律通过非视觉光感应机制。 该计划包括三个主要目标。目的1，生物化学表征，通过 HPLC和质谱法分析神经内分泌调节肽 睫状上皮中表达的高血压/水肿活性， 包括神经肽Y和利钠肽C。来研究它们的信号 通过测量第二信号通路， 信使包括cAMP、cGMP和Ca 2+。目的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 睫状体上皮进入光/暗周期。识别 内源性“昼夜节律振荡器”的组成部分， 睫状上皮中的基因表达，包括褪黑激素节律产生 酶和时钟基因。目的3，确定 睫状体上皮中的非视觉光传导级联，通过聚焦 视紫红质和其他视蛋白样分子的表达， 视紫红质的活化和失活组分， 包括视紫红质激酶、恢复蛋白和抑制蛋白。以确定任何 在完整的和培养的睫状上皮细胞的光敏感反应，通过 研究在光和暗条件下cGMP门控通道。

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.