C elegans cell cycle-dependent C1C-2 channel ortholog

描述（由申请人提供）：C1 C阴离子通道几乎在 所有的有机体。虽然大多数识别的CIC的功能是模糊的， CIC基因存在于广泛不同的生物体中， 人类和其他哺乳动物中的致病CIC突变表明， 经络发挥着重要的生理作用。 C. elegans为描述CIC提供了显著的实验优势 阴离子通道生物学我们证明了C.线虫卵母细胞表达a 哺乳动物CIC-2通道直系同源物，由clh-3编码，clh-3是六种线虫CIC之一 基因. CLH-3被肿胀激活，但对卵母细胞体积没有影响 控制体积敏感性似乎将通道活性与卵母细胞生长联系起来 发展先行者的要求在发育成熟的卵母细胞中，CLH-3是一个重要的调节因子。 组成性激活。卵母细胞成熟诱导排卵收缩， 电耦合鞘细胞。clh-3基因表达的RNA干扰 破坏了鞘收缩的时间，表明通道 通过卵母细胞鞘细胞间信号传导途径调节排卵。 因此，CLH-3作为细胞周期传感器发挥作用，以确保细胞周期的同步化。 排卵和受精的成熟。 该提案的中心重点是确定CLH-3的调节机制 并定义通道在细胞间信号传导途径中的作用。 具体而言，我们将描述卵母细胞生长，卵母细胞细胞 周期进程和受精在调节CLH-3活性方面的作用， 假设细胞周期依赖性激酶调节CLH-3，并测试 推测β-淀粉样蛋白3通过介导鞘细胞钙信号通路调节鞘细胞钙信号通路 卵母细胞去极化和鞘细胞膜电位。结果进行 这些研究对理解人类生理学具有重要意义， 病理生理学拟议的调查将继续扩大我们的 特别是对C1 C-2和一般C1 C阴离子通道的理解。等 为了识别C1 C通道的功能，理解是必不可少的， 它们的调节机制及其作为治疗靶点的潜力， 囊性纤维化等疾病。此外，我们的研究可能会提供 对卵母细胞发育、卵母细胞 细胞周期控制、细胞间通讯机制和激动剂诱导的 平滑肌收缩和调节。

DESCRIPTION (provided by applicant): C1C anion channels are found in virtually all organisms. While the functions of most identified CICs are obscure, the presence of CIC genes in widely divergent organisms and the existence of disease-causing CIC mutations in humans and other mammals indicate that the channels play important physiological roles. C. elegans offers significant experimental advantages for characterizing CIC anion channel biology. We have demonstrated that C. elegans oocytes express a mammalian CIC-2 channel ortholog encoded by clh-3, one of six nematode C1C genes. CLH-3 is activated by swelling, but plays no role in oocyte volume control. Volume sensitivity appears to link channel activity to oocyte growth and development. In full-grown oocytes undergoing rneiotic maturation, CLH-3 is constitutively activated. Oocyte maturation induces ovulatory contractions of electrically-coupled sheath cells. RNA interference of clh-3 expression disrupts the timing of sheath contractions indicating that the channel modulates ovulation via oocyte-sheath cell intercellular signaling pathways. CLH-3 thus functions as a cell cycle sensor to ensure synchronization of maturation with ovulation and fertilization. The central focus of this proposal is to identify CLH-3 regulatory mechanisms and define the role of the channel in cell-to-cell signaling pathways. Specifically, we will characterize the roles of oocyte growth, oocyte cell cycle progression and fertilization in regulating CLH-3 activity, test the hypothesis that cell cycle-dependent kinases regulate CLH-3, and test the hypothesis that CLR-3 modulates sheath cell Ca2+ signaling pathways via depolarization of oocyte and sheath cell membrane potential. Results of these studies have significant implications for understanding human physiology and pathophysiology. Proposed investigations will continue to broaden our understanding of C1C-2 specifically, and of C1C anion channels in general. Such understanding is essential in order to identify the functions of C1C channels, their regulatory mechanisms, and their potential as therapeutic targets for diseases such as cystic fibrosis. In addition, our studies will likely provide new insights into the fundamental problems of oocyte development, oocyte cell cycle control, cell-to-cell communication mechanisms, and agonist-induced smooth muscle contraction and regulation.