Functional Analysis of Sleep Homeostasis in Drosophila

果蝇睡眠稳态的功能分析

• 批准号: 7686447
• 负责人: PAUL J SHAW
• 金额: $ 5万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7686447
• 关键词: Acceleration; Adverse effects; Affect; Animals; Attenuated; Behavioral Assay; Biological Assay; Biological Markers; Cells; Chronic; Condition; Coronary heart disease; Data; Disease; Drosophila genus; Endocrine; Exhibits; Genes; Genetic; Genomics; Goals; Health; Homeostasis; Hour; Human; Immunology; Individual; Insulin; Lead; Lipids; Localized; Long-Term Effects; Mammals; Measures; Messenger RNA; Metabolic syndrome; Metabolism; Modification; Molecular; Molecular Profiling; Morbidity - disease rate; Names; Non-Insulin-Dependent Diabetes Mellitus; Numbers; Obesity; Oligonucleotides; Pathology; Physiology; Play; Polymerase Chain Reaction; Predisposition; Principal Investigator; Process; Rattus; Regulator Genes; Resistance; Role; Signal Transduction; Sleep; Sleep Deprivation; Starvation; System; Testing; Time; Toxic effect; Transgenic Organisms; Wakefulness; Work; awake; blood glucose regulation; cDNA Arrays; circadian pacemaker; cost; energy balance; fly; genetic analysis; interest; loss of function; mortality; mutant; novel; programs; protein expression; research study; response; stressor; vector

DESCRIPTION (provided by applicant): Sleep is controlled by two processes: a homeostatic drive that increases during waking and dissipates during sleep and a circadian pacemaker that controls its timing. Although these two systems can operate independently recent studies suggest a more intimate relationship. Indeed, none has been as dramatic as that found for the canonical loss-of-function clock mutant cycle (cyc01). cyc01 mutants showed a disproportionately large sleep rebound and died following 10 hours of sleep deprivation, although they were more resistant than other clock mutants to various stressors. Our data indicate that the pathology is characterized by an acceleration of the detrimental effects of waking and furthermore, suggests that these processes subsequently increase the need for sleep (Shaw et al., 2002). Using genomic studies in these uniquely sensitive animals, we have begun to identify functional targets of sleep homeostasis and its molecular mechanisms. Thus we know of 100 genes that are modulated by prolonged wakefulness. In order to more fully understand the role these genes play, we will determine their temporal dynamics in response to increasing amounts of waking. Moreover, because we have developed independent genetic, pharmacological and behavioral assays that produce periods of waking that differentially activate homeostatic responses, we will determine the extent to which these genes are specifically associated with homeostasis. More importantly, we have acquired over 110 mutant lines representing approximately 60 of the 100 genetic loci of interest and have begun to evaluate their sleep parameters and responses to sleep deprivation. We propose to characterize select genes further by localizing mRNA and protein expression and to manipulate the activity of these genes and the cells that express them by creating a variety of useful transgenic lines using UAS, GAL4 and GAL80 vectors.

描述（由申请人提供）：睡眠由两个过程控制：在清醒时增加并在睡眠期间消散的稳态驱动力和控制其时间的昼夜节律起搏器。尽管这两个系统可以独立运行，但最近的研究表明它们之间的关系更为密切。事实上，没有一个像典型的功能丧失时钟突变体周期（cyc01）那样引人注目。 cyc01突变体表现出不成比例的大睡眠反弹，并在睡眠剥夺10小时后死亡，尽管它们比其他时钟突变体对各种压力源具有更强的抵抗力。我们的数据表明，病理学的特点是醒来的有害影响加速，此外，表明这些过程随后增加了对睡眠的需求（Shaw 等人，2002）。通过对这些独特敏感动物的基因组研究，我们已经开始确定睡眠稳态的功能目标及其分子机制。因此，我们知道有 100 个基因受到长时间清醒的调节。为了更全面地了解这些基因所发挥的作用，我们将确定它们响应不断增加的觉醒次数的时间动态。此外，由于我们已经开发了独立的遗传、药理学和行为测定法，这些测定法可以产生不同程度地激活稳态反应的觉醒周期，因此我们将确定这些基因与稳态特异性相关的程度。更重要的是，我们已经获得了超过 110 个突变系，代表了 100 个感兴趣的基因位点中的大约 60 个，并已开始评估它们的睡眠参数和对睡眠剥夺的反应。我们建议通过定位 mRNA 和蛋白质表达来进一步表征所选基因，并通过使用 UAS、GAL4 和 GAL80 载体创建各种有用的转基因系来操纵这些基因和表达它们的细胞的活性。