Function of SLEEPLESS in sleep regulation

SLEEPLESS的睡眠调节功能

基本信息

批准号：
8536317
负责人：
Kyunghee Koh
金额：
$ 26.39万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-25 至 2015-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Function of SLEEPLESS in Sleep Regulation Summary Sleep serves essential biological functions, and is conserved from flies to humans. The overall goal of the proposed research is to elucidate the molecular genetic basis of sleep regulation. Through an unbiased forward-genetic screen for short-sleeping mutant flies, we identified a novel gene, sleepless (sss), which is required for normal sleep as well as recovery sleep after deprivation. sss encodes a small, brain-enriched protein expressed on the cell surface. Expression of the Shaker (Sh) potassium channel, previously shown to be important for sleep regulation, is markedly reduced in sss mutants. Several key questions need to be addressed to understand how SSS regulate sleep. Does Sh mediate the effect of sss on sleep? Does SSS act directly on Sh by forming a complex or does it act indirectly via a signaling pathway? Does SSS act on Sh cell-autonomously or non-cell-autonomously? Do circadian and homeostatic processes affect SSS and Sh expression? To address these questions, we propose the following specific aims: (1) Test whether SSS promotes sleep by acting on Sh and whether they physically interact. To confirm a potential genetic interaction between sss and Sh, we will look for a synergistic effect on sleep of reducing both sss and Sh activity. We will also examine whether overexpression of Sh can rescue the short-sleeping phenotype of sss mutants. In addition, we will test whether SSS and Sh physically interact. (2) Identify the locus of action of sss. Identification of sss, which has profound effects on both baseline and rebound sleep, allows us an opportunity to explore the cellular substrates of homeostatic sleep regulation. We will examine the expression pattern of the SSS protein, and carry out rescue experiments using a collection of tissue-specific drivers and an sss transgene. Cellular substrates of sleep under normal and deprived conditions will be compared. (3) Determine if SSS acts on Sh cell-autonomously. To understand how SSS acts on Sh to regulate sleep, it is important to determine the locus of action of Sh as well as that of sss. We will determine the Sh expression pattern and tissue requirement using tissue-specific rescue and targeted RNAi knockdown experiments. In addition, we will test whether SSS can be secreted and whether secreted SSS can rescue the sleep phenotype of sss mutants. PUBLIC HEALTH RELEVANCE: Lack of good-quality sleep is a common problem that can lead to accidents and reduced productivity, and there is growing awareness among healthcare professionals and the public that sleep is a critical health issue. Elucidation of molecular mechanisms underlying sleep regulation may provide us with important clues as to how and why we sleep, and may ultimately lead to novel treatments for sleep disorders.

描述（由申请人提供）： SLEEPLESS 在睡眠调节中的功能摘要睡眠具有重要的生物学功能，从果蝇到人类都具有睡眠功能。该研究的总体目标是阐明睡眠调节的分子遗传基础。通过对短睡眠突变果蝇进行无偏倚的前向遗传筛选，我们发现了一种新的基因——sleepless (sss)，它是正常睡眠以及睡眠剥夺后恢复睡眠所必需的。 sss 编码一种在细胞表面表达的小型大脑富集蛋白质。 Shaker (Sh) 钾通道的表达先前被证明对睡眠调节很重要，但在 sss 突变体中显着降低。要了解 SSS 如何调节睡眠，需要解决几个关键问题。 Sh 是否介导 sss 对睡眠的影响？ SSS 是通过形成复合物直接作用于 Sh 还是通过信号通路间接作用？ SSS 是细胞自主作用还是非细胞自主作用于 Sh？昼夜节律和稳态过程会影响 SSS 和 Sh 表达吗？为了解决这些问题，我们提出以下具体目标：（1）测试SSS是否通过作用于Sh来促进睡眠以及它们是否存在物理相互作用。为了确认 sss 和 Sh 之间潜在的遗传相互作用，我们将寻找减少 sss 和 Sh 活性对睡眠的协同效应。我们还将研究 Sh 的过度表达是否可以挽救 sss 突变体的短睡眠表型。此外，我们还将测试SSS和Sh是否有物理交互。 (2)确定sss的作用位点。 sss 的识别对基线睡眠和反弹睡眠都有深远的影响，使我们有机会探索稳态睡眠调节的细胞基础。我们将检查 SSS 蛋白的表达模式，并使用一系列组织特异性驱动程序和 sss 转基因进行救援实验。将比较正常和剥夺条件下睡眠的细胞基质。 (3)确定SSS是否自主作用于Sh细胞。为了了解 SSS 如何作用于 Sh 来调节睡眠，确定 Sh 和 sss 的作用位点很重要。我们将使用组织特异性救援和靶向 RNAi 敲低实验来确定 Sh 表达模式和组织需求。此外，我们将测试SSS是否可以分泌，以及分泌的SSS是否可以挽救sss突变体的睡眠表型。公共健康相关性：缺乏优质睡眠是一个常见问题，可能导致事故和生产力下降，医疗保健专业人员和公众越来越意识到睡眠是一个重要的健康问题。阐明睡眠调节的分子机制可能为我们提供有关睡眠方式和原因的重要线索，并可能最终导致睡眠障碍的新治疗方法。