DIFFERENTIAL GENE EXPRESSION IN CILIARY EPITHELIUM

描述：眼睫状上皮是神经上皮的双层 具有特征性神经内分泌表型的细胞。 表达式由 编码神经肽加工酶的基因的睫状上皮， 神经肽、肽类激素和神经内分泌受体 组织建立了我们观察的分子基础， 组织显示神经内分泌功能。 PI提出了假设 在眼睛的前段，睫状上皮是 神经肽和肽激素合成的主要位点（即， 神经降压素、心房利钠肽、血管紧张素原）的已知的 心血管系统中的高血压/水肿作用， 调节眼压的潜在作用。 他建议， 睫状上皮通过内分泌机制调节眼压 （自分泌/旁分泌）通过调节肽作用于流入 系统[无色素（NPE）和色素睫状上皮细胞（PE）]， 和/或在流出系统[小梁网细胞（TM）]上。 本研究的主要实验方法是：1）调查 在人睫状体上皮中， 生物活性物质的基因表达、合成、加工和信号通路 在高血压/低血压中具有假定作用的肽及其受体 在正常和青光眼眼中。 2)使用已建立的细胞系， 从流入系统（即，NPE，PE）和流出（即，小梁 网络），作为体外细胞模型系统来定义神经内分泌 睫状上皮的特性。 3)减法的应用 通过杂交鉴定人睫状体中的新候选基因 与高眼内压/低眼压性青光眼相关。 具体 目的如下：1）确定表达的规则 内切蛋白酶（激素原转化酶）和外肽酶（羧肽酶 和肽酰甘氨酸（-酰胺化单加氧酶）参与的加工， 神经肽和肽激素; 2）表征 具有高血压/肥胖作用的调节肽及其受体; 3)确定参与成熟的调节机制， 正常人和青光眼患者的加工酶和神经肽分泌 睫状体上皮细胞; 4）确定信号转导 参与调节的受体激活后的途径 NPE/PE和TM细胞中的肽; 5）确定结构-功能 编码TIGR的人睫状体消减克隆， 一种与青光眼（青少年开角型青光眼）有关的蛋白质， 一个调节眼压的候选基因

DESCRIPTION: The ocular ciliary epithelium is a bilayer of neuroepithelial cells with a characteristic neuroendocrine phenotype. The expression by the ciliary epithelium of genes encoding neuropeptide processing enzymes, neuropeptides, peptide hormones and receptors restricted to neuroendocrine tissues establishes the molecular basis of our observations that this ocular tissue displays neuroendocrine functions. The PI formulated the hypothesis that, in the anterior segment of the eye, the ciliary epithelium is the primary site of synthesis of neuropeptides and peptide hormones (i.e., neurotensin, atrial natriuretic peptide, angiotensinogen) of known hypertensive/hypotensive actions in the cardiovascular system, and with potential roles in the regulation of eye pressure. He proposes that the ciliary epithelium modulates eye pressure by endocrine mechanisms (autocrine/paracrine) through regulatory peptides acting on the inflow system [nonpigmented (NPE) and pigmented ciliary epithelial cells (PE)], and/or on the outflow system [trabecular meshwork cells (TM)]. The major experimental approach of this research will be: 1) to investigate in the human ciliary epithelium, the molecular mechanism involved in the gene expression, synthesis, processing and signaling pathways of bioactive peptides and its receptors with putative roles in hypertension/ hypotension in normal and glaucoma eyes. 2) The use of established cells lines derived from the inflow system (i.e., NPE, PE) and outflow (i.e., trabecular meshwork), as in vitro cell model systems to define the neuroendocrine properties of the ciliary epithelium. 3) The application of subtractive hybridization to identify novel candidate genes in the human ciliary body associated to high intraocular pressure/low tension glaucoma. The specific aims are as follows: 1) Determine the regulation of the expresssion of endoproteases (prohormone convertases), and exopeptidases (carboxypeptidases and peptidylglycine (-amidating monooxygenase) involved in the processing of neuropeptides and peptide hormones; 2) Characterize the gene expression of regulatory peptides with hypertensive/hypotensive actions and its receptors; 3) Determine the regulatory mechanism involved in the maturation and secretion of processing enzymes and neuropeptides in normal and glaucoma derived ciliary epithelial cells; 4) Determine the signal transduction pathways involved upon activation of the receptors for the regulatory peptides in NPE/PE and TM cells; 5) Determine the structure-function relationship of a human ciliary body subtracted clone encoding TIGR, a protein involved in a subset of glaucoma (Juvenile Open Angle Glaucoma) and a candidate gene to regulate eye pressure.