Dissecting the cAMP-mediated circuitry of stress-induced sleep in Caenorhabditis elegans

项目/摘要 脊椎动物和无脊椎动物的睡眠由保守的信号分子控制，这表明睡眠 在进化上是古老的。秀丽线虫为快速鉴定和鉴定提供了强有力的手段 睡眠调节通路的单细胞分辨率特征，因为它们的产生时间很快 以及一个简单而明确的302细胞神经系统。一种被证明可以调节所有动物睡眠的途径 涉及环磷酸腺苷(CAMP)，一种普遍存在的第二信使，主要作用于 神经元。线虫睡眠对环境侮辱引起的细胞压力的反应(压力诱导 睡眠)，这是一个有趣的研究疾病期间睡眠的行为模型。该项目将定义在哪里露营 在压力诱导的睡眠中在线虫的神经系统中发挥作用。它还将确定地点和 压力和睡眠是如何在神经系统中相交的。在目标1中，我们将确定cAMP所在的神经元 并测量活体动物在睡眠时cAMP水平的变化。为了实现这一目标，我们将 使用红光激活的腺酰环化酶在体内允许细胞特异性地诱导cAMP，我们将使用 一种生物传感器，可以实时测量睡眠期间cAMP的变化。在目标2中，我们将测试假设 主应激感受器一磷酸腺苷激活的蛋白激酶(AMPK)在相同的 作为cAMP的神经元，在蛋白激酶A的下游，在睡眠中被激活，以响应细胞压力。 在目标3中，我们将把这些新发现的睡眠神经元与已知的应激诱导睡眠回路联系起来。 并验证AMPK在PKA下游发挥作用的假设。

Project/Summary Vertebrate and invertebrate sleep are controlled by conserved signaling molecules, which suggests that sleep is evolutionarily ancient. Caenorhabditis elegans provides a powerful means for rapid identification and characterization of sleep-regulating pathways with single cell resolution because of their fast generation time and a simple and defined 302-celled nervous system. One pathway shown to regulate sleep in all animals involves cyclic adenosine monophosphate (cAMP), a ubiquitous second messenger that largely functions in neurons. C. elegans sleep in response to cellular stress caused by environmental insults (stress-induced sleep), an intriguing behavioral model for studying sleep during sickness. This project will define where cAMP is functioning in the C. elegans nervous system during stress-induced sleep. It will also determine where and how stress and sleep intersect in the nervous system. In Aim 1, we will identify the neurons in which cAMP is functioning and measure changes in cAMP levels in live animals while they sleep. To accomplish this, we will use a red light activated adenylyl cyclase that allows for cell-specific induction of cAMP in vivo and we will use a biosensor to measure cAMP changes in real-time during sleep. In Aim 2, we will test the hypothesis that the master stress sensor adenosine monophosphate-activated protein kinase (AMPK) is functioning in the same neurons as cAMP, downstream of protein kinase A, and is activated during sleep in response to cellular stress. In Aim 3 we will connect these newly identified sleep neurons to the known circuitry of stress induced sleep and test the hypothesis that AMPK is functioning downstream of PKA.