HUMAN LENS CONNEXIN GENES IN HEREDITARY CATARACT

遗传性白内障中的人类晶状体连接蛋白基因

• 批准号: 6179043
• 负责人: Alan Shiels
• 金额: $ 23.82万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6179043
• 关键词: HeLa cells; Xenopus oocyte; autosomal dominant trait; cataract; congenital eye disorder; electrophysiology; family genetics; fluorescent dye /probe; gap junctions; gene expression; gene mutation; genetic mapping; genetically modified animals; human genetic material tag; laboratory mouse; membrane channels; molecular cloning; nucleic acid sequence; transfection; voltage /patch clamp

DESCRIPTION: Inherited cataract is a sight-threatening lens disease that usually presents as a congenital, autosomal dominant Mendelian trait showing high penetrance and considerable inter- and intra-familial clinical variation. The primary objective of this project is to determine whether mutations in the genes for connexin50 (Cx50) and connexin46 (CX46) underlie autosomal dominant forms of zonular pulverulent cataract that the PI has mapped to human chromosomes 1q and 13q, respectively. DNA cloning and sequencing techniques will be used to identify mutations and engineer constructs for functional expression studies on the wild-type and mutant forms of Cx50 and Cx46. The voltage-gating properties of wild-type and mutant forms Cx50 and Cx46 synthesized in Xenopus oocytes will be determined using a two microelectrode voltage clamp technique under defined pH conditions. The intercellular channel properties of wildtype and mutant forms Cx50 and Cx46 expressed in the connexin-deficient HeLa cell-line will be measured by fluorescent dye-transfer methods. Transgenic mouse techniques will be used to model the allelic interactions of wild-type and mutant forms of Cx50 and Cx46 that lead to cataract development in the living lens. Results from these studies will help to elucidate the pathogenetic mechanisms underlying a clinically important form of inherited cataract in humans and provide new insight into the role of connexins in human lens development. Ultimately, such data will contribute to the design of new preventative and therapeutic strategies for the clinical management of cataract.

描述：遗传性白内障是一种威胁视觉镜头疾病 通常以先天性，常染色体主导的孟德尔特征而呈现 高渗透率高以及大量的家庭间临床和大量 变化。 该项目的主要目的是确定是否 Connexin50（CX50）和Connexin46（CX46）的基因突变 Zonular粉状白内障的常染色体显性形式，PI具有 分别映射到人类染色体1q和13q。 DNA克隆和测序技术将用于识别突变和 用于野生型和野生型功能表达研究的工程构造 CX50和CX46的突变形式。 野生型的电压门控性能 在异爪蟾卵母细胞中合成的突变体形式的CX50和CX46将是 使用定义的两个微电极电压夹技术确定 pH条件。 野生型和突变体的细胞间通道特性 缺乏连接蛋白的HELA细胞系中表达的CX50和CX46形式将 通过荧光染料转移方法测量。 转基因小鼠 技术将用于建模野生型和 CX50和CX46的突变形式导致白内障发育 活镜头。 这些研究的结果将有助于阐明 遗传上临床上重要形式的致病机制 人类白内障，并提供有关连接素在 人镜头发展。 最终，此类数据将有助于设计 临床管理的新预防和治疗策略 白内障。