卤虫休眠胚胎（卵）通道蛋白解析及其对休眠的调控作用研究

卤虫是生活在盐湖盐田等极端环境中的小型甲壳动物，通过产休眠卵的生殖方式适应寒冷高盐等胁迫环境。对卤虫休眠胚胎形成和细胞静息调控机制的研究一直是甲壳动物学的重点内容。迄今，人们对环境因子调控休眠胚胎（卵）形成和细胞静息的分子机制了解甚少。细胞膜表面离子通道是介导环境因子与细胞互作的重要环节，但是，关于卤虫休眠胚胎离子通道的研究尚属空白。我们前期发现卤虫休眠胚胎（卵）细胞膜存在明显电位差，提示膜通道蛋白参与卤虫休眠胚胎形成和细胞静息调控。本项目拟采用全细胞膜片钳和分子克隆技术，结合特异性激动剂和阻断剂的使用，解析卤虫休眠胚胎细胞表面通道蛋白种类和电生理特征，观察胁迫环境因子对通道蛋白表达和电生理特征的影响，阐明1-2种通道蛋白在卤虫休眠胚胎形成和细胞静息调控中的作用，解析其激活的细胞信号通路，探讨离子通道介导环境信号调控卤虫胚胎休眠的分子机制，为了解和控制动物休眠和细胞静息调控机制奠定基础。

Salt shrimp (Artemia salina) is a small crustacean which live in extreme environment such as salt lakes and salt pans. In response to environmental stress, such as cold and high-salinity, Artemia produce embryos that rapidly enter a state of dormancy, called diapause. The research on the regulation of diapause embryos and cellular quiescence has been the focus of crustacean biology, However, the molecular mechanism of environmental factors regulating the diapause formation of Artemia has not been clearly studied up to date. We have established a stress-induced model of ovoviviparous and oviparous, our previous studies showed that in the process of the initiation of the diapause cysts, the change of membrane potential was dependent on environment change. These results suggest that channel proteins may be a factor of embryo dormancy and cellular quiescence. In this project, we will analyze the types and characteristics of channel proteins in the cytomembrane of diapause embryos by using whole-cell patch-clamp and molecular cloning, combined with the use of specific agonists and blockers, to study the effects of environmental factors on the expression and electrophysiological characteristics of channel proteins, and to identify the key channel proteins involved in diapause formation and cellular quiescence, then we will focus the secondary messenger and signal pathways activated by the signal of channel protein response to environmental stress, elucidate the molecular regulation mechanism of channel proteins in the formation of diapause embryos and the regulation of cellular quiescence in Artemia. The result of this study will provide a unique idea for the regulation of animal dormancy and cellular quiescence.