MOLECULAR IDENTITY OF A C ELEGANS OOCYTE CL CHANNEL

A C 线虫卵母细胞 CL 通道的分子特性

• 批准号: 6292991
• 负责人: ERIC M RUTLEDGE
• 金额: $ 4.02万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6292991
• 关键词: Caenorhabditis elegans; biological signal transduction; cell cycle; cell growth regulation; cell morphology; chloride channels; complementary DNA; developmental genetics; egg /ovum; electrophysiology; gene expression; gene targeting; genome; mutant; oogenesis; ovulation; phosphorylation; polymerase chain reaction; protein structure function; voltage /patch clamp

The ability to sense and respond to changes in cell volume is a universal property of all cells. Changes in cell volume play important signaling roles in controlling cell growth and proliferation. Cell volume regulation is an essential "housekeeping" function. Despite their physiological importance, little is known about the molecular nature of volume- sensitive channels, or about how cell size is sensed and regulates channel gating. C. elegans offers significant experimental advantages for defining the function and regulation of ion channels. However, because of the relative inaccessibility of somatic cells in adult nematodes, direct electrophysiological studies of ion channel activity are extremely difficult. To circumvent this problem, I developed techniques to patch clamp C. elegans oocytes. My studies have identified a swelling- activated Inwardly Rectifying Cl-Channel termed IRCC. IRCC activity changes dramatically during oocyte development suggesting that the channel may play an important role in regulating oocyte growth and/or the cell cycle. Preliminary RNA interference data suggest that IRCC is encoded by a ClC channel gene. The central goal of my proposal is to combine electrophysiological measurements with genomic analysis and molecular biology methods to identify the gene(s) encoding IRCC and to begin characterizing channel regulation. Identification of the channel genes will ultimately enable them to be manipulated at the molecular level. This in turn will provide insight into the role of IRCC in oogenesis and into the mechanisms by which it senses and is regulated by cell size.

感知和响应细胞体积变化的能力是所有细胞的普遍特性。细胞体积的变化在控制细胞生长和增殖中发挥重要的信号作用。细胞体积调节是一项重要的“管家”功能。尽管它们具有生理重要性，但人们对体积敏感通道的分子性质或如何感知细胞大小并调节通道门控知之甚少。线虫为定义离子通道的功能和调节提供了显着的实验优势。然而，由于成体线虫体细胞相对难以接近，离子通道活性的直接电生理学研究极其困难。为了解决这个问题，我开发了膜片钳线虫卵母细胞的技术。我的研究发现了一种膨胀激活的内向整流 Cl 通道，称为 IRCC。 IRCC 活性在卵母细胞发育过程中发生显着变化，表明该通道可能在调节卵母细胞生长和/或细胞周期中发挥重要作用。初步 RNA 干扰数据表明 IRCC 由 ClC 通道基因编码。我的提案的中心目标是将电生理测量与基因组分析和分子生物学方法相结合，以识别编码 IRCC 的基因并开始表征通道调节。通道基因的鉴定最终将使它们能够在分子水平上被操纵。这反过来将有助于深入了解 IRCC 在卵子发生中的作用及其感知细胞大小和受细胞大小调节的机制。