C elegans cell cycle-dependent C1C-2 channel ortholog

描述(申请人提供)：C1C阴离子通道可在虚拟 所有的有机体。虽然大多数已确定的CIC的功能模糊不清，但 CIC基因在差异很大的生物中的存在和CIC基因的存在 在人类和其他哺乳动物中致病的CIC突变表明 经络起着重要的生理作用。 线虫为表征CIC提供了显著的实验优势 阴离子通道生物学。我们已经证明线虫卵母细胞表达一种 由六种线虫C1C之一的CLH-3编码的哺乳动物CIC-2通道同源基因 基因。CLH-3被肿胀激活，但对卵母细胞体积没有影响 控制力。体积敏感性似乎将通道活动与卵母细胞生长联系在一起 和发展。在成熟的卵母细胞中，CLH-3是 构成激活的。卵母细胞成熟诱导排卵收缩 电耦合鞘电池。RNA干扰对CLH-3表达的影响 扰乱了鞘收缩的时间，表明经络 通过卵母细胞-鞘细胞间信号通路调节排卵。 因此，CLH-3起到了细胞周期传感器的作用，以确保 成熟伴随着排卵和受精。 这项提案的中心焦点是确定CLH-3的调节机制 并定义了该通道在细胞间信号通路中的作用。 具体地说，我们将描述卵母细胞生长、卵母细胞 周期进展和受精在调节CLH-3活性中的作用，测试 假设细胞周期依赖的激酶调节CLH-3，并测试 CLR-3通过调节鞘细胞钙信号通路的假说 卵母细胞去极化和鞘细胞膜电位。这些措施的结果 研究对理解人类生理学和 病理生理学。拟议的调查将继续扩大我们的 对C1C-2的具体了解，以及对C1C阴离子通道的一般了解。是这样的 为了确定C1C通道的功能，理解是必不可少的， 它们的调节机制，以及它们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 对卵母细胞发育基本问题的新认识 细胞周期控制、细胞间通讯机制和激动剂诱导 平滑肌肉的收缩和调节。

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.