THE FUNCTION OF BESTROPHIN IN RETINAL DEGENERATION

Bestrophin 在视网膜变性中的作用

基本信息

批准号：
6294509
负责人：
MARGARET M DEANGELIS
金额：
$ 3.24万
依托单位：
MASSACHUSETTS EYE AND EAR INFIRMARY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-10-01 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/6294509
关键词：
electrooculography electrophysiology immunocytochemistry laboratory mouse macular degeneration membrane proteins microscopy molecular pathology protein localization protein structure function retina degeneration retinal pigment epithelium single strand conformation polymorphism

项目摘要

DESCRIPTION (Applicant's Description): The overall objective of the proposed studies is to obtain information regarding the role of the bestrophin gene (VMD2) in retinal degeneration. Best Macular dystrophy (BMD, also called vitelliform macular dystrophy) is an autosomal dominant form of macular degeneration. BMD, caused by mutations in the VMD2 gene, leads to a progressive loss of central vision. Little is known about the protein product of this gene, called bestrophin. In the retina, bestrophin is specifically expressed in the plasma membrane of the retinal pigment epithelium (RPE). In the following application, I plan to conduct research toward understanding the physiological and molecular role of VMD2 in the retina. The first specific aim seeks to determine if mutations in the VMD2 gene cause other forms of retinal degeneration such as age-related macular degeneration (AMD) using PCR-based single-strand conformation polymorphism and direct genomic sequencing techniques. In the second aim, I plan to ascertain the subcellular localization of bestrophin in the plasma membrane of the RPE using imunocytochemistry. In the last aim, I propose in vitro electrophysiological studies as well as in vivo studies using mice. These studies include methods such as Ussing chambers, light, electron and fluorescent microscopy, gene transfer both in vivo and in vitro, electrocculography, and electroretinography. Discovering information regarding the function of bestrophin will hopefully provide knowledge about mechanisms involved in retinal degeneration.

描述（申请人的描述）：拟议的总体目标研究的目的是获得有关黄斑黄蛋白基因作用的信息 (VMD2) 视网膜变性。最佳黄斑营养不良症（BMD，也称为卵黄样黄斑营养不良）是黄斑部的常染色体显性遗传形式退化。由 VMD2 基因突变引起的 BMD 会导致进行性骨密度下降中央视力丧失。人们对这个基因的蛋白质产物知之甚少，称为斑黄蛋白。在视网膜中，黄斑黄蛋白特异性表达于视网膜色素上皮 (RPE) 的质膜。在下文中应用程序，我计划进行研究以了解生理学以及 VMD2 在视网膜中的分子作用。第一个具体目标旨在确定 VMD2 基因突变是否会导致其他形式的视网膜病变使用基于 PCR 的变性，例如年龄相关性黄斑变性 (AMD) 单链构象多态性和直接基因组测序技术。在第二个目标中，我计划确定亚细胞定位使用免疫细胞化学法检测 RPE 质膜中的 bestropin。在最后一个目标，我建议进行体外电生理学研究以及使用小鼠进行体内研究。这些研究包括使用诸如室、光、电子和荧光显微镜、基因转移体内和体外、眼电图和视网膜电图。发现有关 bestropin 功能的信息有望提供有关视网膜变性机制的知识。