INTRACELLULAR CALCIUM CONTROL OF cAMP SYNTHESIS

描述(改编自申请者的描述)：腺酰环化酶生成 无处不在的第二信使阵营。这位信使在中国的重要性 对大量细胞过程的调控是公认的。然而， 测量cAMP的方法缺乏时间和空间分辨率，导致 因为人们普遍认为这些信号很简单。与此相反的是， 钙离子与cAMP信号通路之间的大量相互作用 已经被证明，而钙离子的动态性质使cAMP很可能 信号也很复杂。特别是，对钙离子敏感的腺酰环化酶 钙离子进入和cAMP调节的生理模式的谨慎调节 Ca~(2+)通过多种机制进入体内。本申请提出了 发展腺酰环化酶/水飞蓟素和腺酰环化酶/骆驼嵌合体 钙离子的定位感受器与嗅觉环核苷酸门控研究 (OCNG)通道作为cAMP的快速膜结合传感器。一旦这些传感器 经过优化，申请者打算检查动态和相互依赖的变化 在这两种信号中，脑垂体来源的GH4细胞是一种可兴奋的细胞类型。他 还将使用cAMP传感器来研究腺酰环化酶的快速动力学 在其本土环境中的监管。这些研究是第一次尝试 在相同的时间和空间尺度上将cAMP和钙信号联系起来。在……里面 从长远来看，这样的信息将导致更广泛的理解 这两个第二信使的协调活动如何控制细胞 功能。

DESCRIPTION (Adapted from applicant's description): Adenylyl cyclases generate the ubiquitous second messenger cAMP. The importance of this messenger in regulating a large number of cellular processes is acknowledged. However, the methods for measuring cAMP lack both temporal and spatial resolution, leading to the widespread belief that these signals are simple. Against this notion, the large number of interactions between signaling pathways for Ca2+ and cAMP have been demonstrated and the dynamic nature of Ca2+ makes it likely that cAMP signals are also complex. In particular, Ca2+-sensitive adenylyl cyclases are regulated discreetly by physiological modes of Ca2+ entry and cAMP modulates Ca2+ entry by a variety of mechanisms. The present application proposes to develop adenylyl cyclase/ aequorin and adenylyl cyclase/ cameleon chimeras as localized sensors of Ca2+ and to develop olfactory cyclic nucleotide gated (oCNG) channels as rapid membrane-bound sensors of cAMP. Once these sensors are optimized, the applicant intends to examine dynamic and interdependent changes in the two signals in pituitary-derived GH4 cells, an excitable cell type. He will also use the cAMP sensor to study the rapid kinetics of adenylyl cyclase regulation in its native environment. These studies represent a first attempt to associate cAMP and Ca2+ signaling on the same temporal and spatial scale. In the long-term, such information will lead to greater generalized understanding of how the coordinated activity of these two second messengers control cellular function.