GAP JUNCTIONAL COMMUNICATION IN LENS FUNCTION

晶状体功能中的间隙连接通讯

• 批准号: 2602725
• 负责人: NORTON B GILULA
• 金额: $ 32.32万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2602725
• 关键词: cataract; cell cell interaction; confocal scanning microscopy; electrophysiology; gap junctions; gel electrophoresis; gene expression; gene mutation; genetic regulation; genetically modified animals; immunocytochemistry; in situ hybridization; laboratory mouse; mass spectrometry; membrane channels; pathologic process; protein structure function

DESCRIPTION: Gap junctions contain channels for the movement of molecular information between cells. These channels are formed by the assembly of products of the gap junction genes called connexins. Communication between cells via these channels provides a mechanism for transferring small molecules that can regulate and synchronize the activities of cells in specialized multi-cellular systems such as the eye lens. Gap junctional communication has been well documented for some time as an important property between cells in the eye lens. Recently, the PI has targeted the disruption of one of the gap junction genes, a3 connexin, that is preferentially expressed in lens fibers using ES cell technology and mouse embryos. This experimental procedure has resulted in the development of an a3 connexin knockout mouse that contains a lens opacity resembling the human age-dependent nuclear cataract phenotype. Thus, this analysis has provided the first direct evidence for an important role for gap junctions in the physiology of the lens. The long term objective of this project is to understand how alterations in the gap junctional communication pathway that result from disruption of a connexin gene contribute to the formation of a cataract. The analysis will include the further characterization of the animal with the knockout of the a3 connexin gene, as well as the generation of an animal with a disruption of another connexin gene, a8, that is used by lens fiber cells. Once both knockout animals have been generated, homozygote mice containing knockouts of both a3 and a8 connexin genes will be analyzed. The connexin knockout mice will be extensively characterized to determine both morphological and biochemical changes that have taken place that contribute to the observed phenotypes. In addition, an experimental strategy will be applied to try and rescue the a3 connexin knockout mice by using the transgenic expression of a8 connexin in the lens. The generation of these knockout mice will also provide a unique opportunity to identify small biologically relevant molecules that may be transmitted via gap junctional communication to influence the process of cataract formation.

描述：间隙连接包含分子运动的通道， 细胞之间的信息。 这些通道由以下组件形成： 称为连接蛋白的差距连接基因的产物。 之间的通信 细胞通过这些通道提供了一种机制， 能够调节和同步细胞活动的分子， 专门的多细胞系统，例如眼睛透镜。 间隙连接通讯作为一种 眼睛透镜中细胞之间的重要性质。 最近，PI 目标是破坏一种差距连接基因a3连接蛋白，该基因 使用ES细胞技术在透镜纤维中优先表达， 小鼠胚胎 这一实验过程导致了 一个a3连接蛋白敲除小鼠，含有一个透镜不透明类似的 人年龄依赖性核性白内障表型。 因此，这一分析 为缝隙连接的重要作用提供了第一个直接证据 在透镜的生理学中。 该项目的长期目标是了解如何改变 差距连接通讯途径，由中断的一个 连接蛋白基因与白内障的形成有关。 该分析将 包括进一步表征敲除了 a3连接蛋白基因，以及具有破坏的动物的产生 另一个连接蛋白基因a8，被透镜纤维细胞使用。 一旦两 已经产生了敲除动物，含有敲除的纯合子小鼠 将分析A3和A8连接蛋白基因的基因。 连接蛋白敲除 小鼠将被广泛地表征以确定形态学和 已经发生的生化变化，有助于观察到的 表型 此外，还将应用一种实验策略， 并通过使用转基因表达挽救A3连接蛋白敲除小鼠， 透镜中的a8连接蛋白。 这些基因敲除小鼠的产生也将 提供了一个独特的机会，以确定小的生物相关的 可以通过间隙连接通讯传递的分子， 影响白内障形成的过程。