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: 2888631
• 负责人: Alan Shiels
• 金额: $ 22.5万
• 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 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2888631
• 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力的晶状体疾病， 通常表现为先天性、常染色体显性孟德尔性状 高外显率和相当多的家庭间和家庭内临床 变化。 该项目的主要目标是确定是否 connexin50 (Cx50) 和 connexin46 (CX46) 基因突变是其基础 PI 患有常染色体显性遗传形式的小带粉状白内障 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 用于野生型和功能表达研究的工程构建体 Cx50 和 Cx46 的突变形式。 野生型的电压门控特性 爪蟾卵母细胞中合成的突变形式 Cx50 和 Cx46 将 使用定义下的两个微电极电压钳技术确定 pH 条件。 野生型和突变体的细胞间通道特性 在连接蛋白缺陷的 HeLa 细胞系中表达的 Cx50 和 Cx46 形式将 通过荧光染料转移方法进行测量。 转基因小鼠 技术将用于模拟野生型和 Cx50 和 Cx46 的突变形式会导致白内障的发生 生活镜头。 这些研究的结果将有助于阐明 临床上重要的遗传形式的发病机制 人类白内障，并为连接蛋白在白内障中的作用提供了新的见解 人类晶状体的发展。 最终，这些数据将有助于设计 临床管理的新预防和治疗策略 白内障。