Interdisciplinary studies of sleep and circadian rhythms in Drosophila

果蝇睡眠和昼夜节律的跨学科研究

• 批准号: 8368866
• 负责人: Michael Warren Young
• 金额: $ 37.08万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-03 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8368866
• 关键词: Advanced Sleep Phase Syndrome; Affect; Behavior; Biochemical; Brain; Cell Count; Cells; Chronic Insomnia; Circadian Rhythms; Clinical; Code; Complement; Complex; Cycas; Development; Drosophila Proteins; Drosophila genus; Fishes; Flow Cytometry; Gene Chips; Gene Expression Profile; Genes; Genetic; Genetic Screening; Goals; Hormones; Human; Inborn Genetic Diseases; Individual; Institutes; Interdisciplinary Study; Investigation; Japan; Libraries; Link; Mitotic; Molecular; Molecular Genetics; Mus; Mutation; Narcolepsy; Nervous system structure; Neurons; Orthologous Gene; Pathway interactions; Pattern; Physiology; Population; Protein Kinase; RNA; RNA Interference; Rana; Rest; Role; Sleep; Sleep Apnea Syndromes; Sleep Disorders; System; Testing; Time; Transgenes; Vertebrates; Wakefulness; Work; X Chromosome; autosome; base; casein kinase I; cdc Genes; circadian pacemaker; economic impact; fly; insight; mutant; nodal myocyte; novel; null mutation; receptor; relating to nervous system; sleep regulation; success; tool; ubiquitin ligase

DESCRIPTION (provided by applicant): Our prior work has focused on the molecular mechanisms underlying Drosophila's circadian rhythms. Orthologs of genes initially characterized in Drosophila have now been linked to the control of rhythmic behavior and physiology in vertebrates, including fish, frogs, mice and humans. Here we propose three classes of interdisciplinary investigations of the fly's rhythmic activity/rest behavior. (1) We wil use elav-Gal4 to drive individual, UAS-RNAi transgenes from two extensive libraries in a search for novel genetic pathways regulating sleep. Our objective is to test the full complement of Drosophila's protein-coding genes using ~19,000 RNAi lines. (2) We will extend our studies of a novel, reduced-sleep mutant, insomniac. We will determine whether insomniac regulates sleep in an active/dynamic manner, or whether it regulates a developmental pathway that is essential for wild type levels of sleep. We will test the hypothesis that Insomniac functions as a substrate-specific adapter for the Cul3 ubiquitin ligase complex, and will use a variety of biochemical and molecular approaches to identify a target substrate(s). (3) We will further define the role of a small group of cell cycle genes, CycA, its regulator (Rca1), cdk2 and cdc42, in the regulation of sleep. As constitutive activation of a cluster of neurons expressing cell cycle genes induces excess sleep, we will isolate these cells by flow cytometry to discover patterns of gene expression that may underlie the contribution of these cells to sleep. PUBLIC HEALTH RELEVANCE: The pressing need to understand sleep is underscored by the major clinical and economic impact of sleep disorders~ 10-15 percent of the U.S. population suffers from chronic insomnia, and nearly one hundred sleep disorders have been described, including narcolepsy, sleep apnea, and delayed and advanced sleep phase syndromes. Molecular genetic studies that began in Drosophila have already allowed mutant orthologs of Period protein and casein kinase 1 to be connected to inborn errors of human sleep. We believe our proposed genetic, cellular, and biochemical studies of Drosophila's activity/rest behavior will continue to provide insights and tools relevant to the understanding of human sleep.

描述（由申请人提供）：我们先前的工作集中在果蝇昼夜节律的分子机制。最初在果蝇中表征的基因的直系同源物现在已经与脊椎动物（包括鱼、青蛙、小鼠和人类）的节律行为和生理学的控制相关联。在这里，我们提出了三个类的跨学科调查苍蝇的节奏活动/休息行为。(1)我们将使用elav-Gal 4驱动来自两个广泛库的单个UAS-RNAi转基因，以寻找调节睡眠的新型遗传途径。我们的目标是使用~ 19，000个RNAi株系来测试果蝇蛋白质编码基因的完整性。(2)我们将扩展我们对一种新的睡眠减少突变体失眠症患者的研究。我们将确定失眠症患者是否以主动/动态的方式调节睡眠，或者它是否调节对野生型睡眠水平至关重要的发育途径。我们将测试Insomniac作为Cul 3泛素连接酶复合物的底物特异性衔接子的假设，并将使用各种生物化学和分子方法来鉴定靶底物。(3)我们将进一步确定一小组细胞周期基因，CycA，其调节因子（Rca 1），cdk 2和cdc 42在睡眠调节中的作用。由于表达细胞周期基因的神经元簇的组成性激活诱导过度睡眠，我们将通过流式细胞术分离这些细胞，以发现可能构成这些细胞对睡眠的贡献的基因表达模式。 公共卫生相关性：睡眠障碍的主要临床和经济影响强调了了解睡眠的迫切需要-10- 15%的美国人口患有慢性失眠症，并且已经描述了近100种睡眠障碍，包括嗜睡症，睡眠呼吸暂停以及延迟和晚期睡眠综合征。从果蝇开始的分子遗传学研究已经允许Period蛋白和酪蛋白激酶1的突变直系同源物与人类睡眠的先天性错误有关。我们相信，我们提出的果蝇活动/休息行为的遗传、细胞和生化研究， 继续提供与了解人类睡眠相关的见解和工具。