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

剖析秀丽隐杆线虫应激诱导睡眠的 cAMP 介导回路

• 批准号: 9231668
• 负责人: Matthew D Nelson
• 金额: $ 32.4万
• 依托单位: SAINT JOSEPH'S UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-20 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9231668
• 关键词: Accidents; Adenosine Monophosphate; Adenylate Cyclase; Affect; Animal Model; Animals; Antibodies; Behavior; Behavioral Model; Biosensor; Caenorhabditis elegans; Calmodulin; Camping; Catalytic Domain; Cells; Cellular Stress; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Drosophila genus; Drosophila melanogaster; Exposure to; Fluorescence Resonance Energy Transfer; GTP-Binding Protein alpha Subunits, Gs; Generations; Genetic Transcription; Growth; Human; Infection; Interneurons; Invertebrates; Lead; Life; Light; Mammals; Measures; Mediating; Mental Health; Metabolic Activation; Metabolism; Modeling; Molecular; Mutation; Nature; Nematoda; Nervous system structure; Neurons; Neuropeptides; Outcome; Pathway interactions; Phosphorylation; Phosphotransferases; Phylogeny; Play; Protein Kinase; Public Health; Resolution; Resources; Role; Second Messenger Systems; Signal Transduction; Signaling Molecule; Sleep; Sleep Disorders; Sleep Wake Cycle; Stress; System; Testing; Time; Translations; Wakefulness; Western Blotting; Work; base; extreme temperature; in vivo; mutant; neural circuit; neuronal excitability; new therapeutic target; optogenetics; physical conditioning; promoter; receptor coupling; repaired; response; restoration; second messenger; sensor; sleep regulation

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.

项目/摘要 脊椎动物和无脊椎动物的睡眠是由保守的信号分子控制的，这表明睡眠 在进化上是古老的。秀丽隐杆线虫为快速鉴定和 由于其快速生成时间，使用单细胞分辨率表征睡眠调节途径 和简单而明确的302细胞神经系统。一种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C.线虫睡眠是对由环境损伤引起的细胞应激的反应（应激诱导的 sleep），这是一个有趣的行为模型，用于研究疾病期间的睡眠。该项目将定义cAMP 在C区运作。神经系统在压力诱导的睡眠。它还将决定在哪里和 压力和睡眠如何在神经系统中相互作用在目标1中，我们将确定cAMP在其中的神经元。 功能和测量的变化cAMP水平在活的动物，而他们的睡眠。为实现这一目标，我们将 使用红光激活的腺苷酸环化酶，其允许在体内细胞特异性诱导cAMP，我们将使用 一种生物传感器，用于测量睡眠期间cAMP的实时变化。在目标2中，我们将检验以下假设： 主应激传感器腺苷一磷酸活化蛋白激酶（AMPK）在相同的 在神经元中，cAMP是蛋白激酶A的下游，并且在睡眠期间响应于细胞应激而被激活。 在目标3中，我们将把这些新发现的睡眠神经元与已知的压力诱导睡眠回路连接起来。 并检验AMPK在PKA下游发挥作用的假设。