Interdisciplinary studies of sleep and circadian rhythms in Drosophila

果蝇睡眠和昼夜节律的跨学科研究

基本信息

批准号：
8463631
负责人：
Michael Warren Young
金额：
$ 35.78万
依托单位：
ROCKEFELLER UNIVERSITY
依托单位国家：
美国
项目类别：
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/8463631
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

项目摘要

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.

描述（由申请人提供）：我们之前的工作重点是果蝇昼夜节律的分子机制。最初在果蝇中鉴定的基因直向同源物现在已与脊椎动物（包括鱼、青蛙、小鼠和人类）的节律行为和生理学的控制联系起来。在这里，我们提出了对果蝇的节律性活动/休息行为进行三类跨学科研究。 (1) 我们将使用 elav-Gal4 驱动来自两个广泛库的个体 UAS-RNAi 转基因，以寻找调节睡眠的新遗传途径。我们的目标是使用约 19,000 个 RNAi 系来测试果蝇蛋白质编码基因的完整互补。 (2) 我们将扩展对一种新型睡眠减少突变体——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 Cul3 泛素连接酶复合物的底物特异性接头的假设，并将使用各种生化和分子方法来识别目标底物。 (3)我们将进一步明确一小群细胞周期基因CycA及其调节因子(Rca1)、cdk2和cdc42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会诱导过度睡眠，因此我们将通过流式细胞术分离这些细胞，以发现可能是这些细胞对睡眠贡献的基因表达模式。