THE FUNCTION OF BESTROPHIN IN RETINAL DEGENERATION

描述(申请人描述)：建议的总体目标 研究是为了获得关于Bestrophin基因作用的信息 (VMD2)在视网膜变性中。最佳黄斑营养不良(BMD，又称 卵黄样黄斑营养不良)是一种常染色体显性黄斑类型 退化。由VMD2基因突变引起的BMD导致进行性 中心视力丧失。人们对该基因的蛋白质产物知之甚少， 叫贝斯特罗芬。在视网膜中，Bestrophin在 视网膜色素上皮(RPE)的质膜。在以下内容中 应用，我计划进行研究，以了解生理 以及VMD2在视网膜中的分子作用。第一个具体目标是寻求 确定VMD2基因突变是否会导致其他形式的视网膜 基于聚合酶链式反应的老年性黄斑变性(AMD)等变性 单链构象多态性与基因组直接测序 技巧。在第二个目标中，我计划确定亚细胞定位 用免疫细胞化学方法检测视网膜色素上皮细胞质膜中的Bestrophin。在……里面 最后一个目的，我建议进行体外电生理研究，以及 用小鼠进行活体研究。这些研究包括Ussing等方法 小室，光，电子和荧光显微镜，基因转移 活体和体外、心电图术和视网膜电图术。发现 有关Bestrophin功能的信息有望提供 有关视网膜变性机制的知识。

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.