THE FUNCTION OF BESTROPHIN IN RETINAL DEGENERATION

Bestrophin 在视网膜变性中的作用

• 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
• 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; 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

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）：提议的总体目标 研究是为了获得有关Bestrophin基因作用的信息 （VMD2）视网膜变性。最佳黄斑营养不良（BMD，也称为 Vitelliform黄斑营养不良症）是一种常染色体显性形式的黄斑形式 退化。由VMD2基因突变引起的BMD导致渐进性 失去中央视力。关于该基因的蛋白质产物知之甚少 称为bestrophin。在视网膜中，bestrophin在 视网膜色素上皮（RPE）的质膜。在以下 应用，我计划进行研究以了解生理 VMD2在视网膜中的分子作用。第一个特定目标试图 确定VMD2基因中的突变是否引起其他形式的视网膜 使用基于PCR的年龄相关黄斑变性（AMD）等变性 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 RPE的质膜中的Bestrophin使用毫米细胞化学。在 最后一个目的，我提出了体外电生理研究以及 使用小鼠的体内研究。这些研究包括诸如USSing之类的方法 腔室，光，电子和荧光显微镜，基因转移都在 体内和体外，电摄影和视网膜造影。发现 有关Bestrophin功能的信息将有望提供 有关视网膜变性涉及的机制的知识。