Analysis of EGFR-dependent sleep in C. elegans

线虫 EGFR 依赖性睡眠分析

• 批准号: 8472826
• 负责人: CHERYL LYNN VAN BUSKIRK
• 金额: $ 14.5万
• 依托单位: CALIFORNIA STATE UNIVERSITY NORTHRIDGE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-10 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8472826
• 关键词: Animal Model; Behavior; Behavioral; Behavioral Assay; Biological Assay; Biological Clocks; Biological Models; Caenorhabditis elegans; Cardiovascular Diseases; Chronic; Circadian Rhythms; Cognitive; Collaborations; Complex; Cues; Data; Development; Developmental Process; Diabetes Mellitus; Disease; Electroencephalogram; Elements; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Ethers; Genes; Genetic; Genetic Techniques; Goals; Homologous Gene; Human; Hypertension; Investigation; Ion Channel; Lead; Ligands; Light; Link; Lobular Neoplasia; Mammals; Mediating; Modeling; Molecular; Molecular Genetics; Molting; Movement; Nematoda; Nervous system structure; Neurons; Neuropeptides; Neurosecretion; Neurosecretory Systems; Obesity; Organism; Orthologous Gene; Pathway interactions; Pattern; Peptide Hydrolases; Performance; Periodicity; Potassium Channel; Process; Property; Publishing; RNA; RNA Interference; RNA Splicing; Regulation; Research; Resistance; Risk; Role; Signal Transduction; Sleep; Sleep Disorders; Staging; System; Testing; Time; Transmembrane Domain; Vertebrates; Voltage-Gated Potassium Channel; Work; base; cognitive function; functional genomics; insight; mutant; neuronal excitability; novel; obesity risk; programs; public health relevance; response; sleep regulation; tool

DESCRIPTION (provided by applicant): Disordered sleep impairs cognitive performance, and chronic sleep loss is associated with increased risk of cardiometabolic diseases including obesity and hypertension. Despite the fundamental importance of sleep, its function remains controversial and the molecular mechanisms by which it is executed are only beginning to be elucidated. Sleep is recognized to be regulated by conserved genetic mechanisms, and in the past decade the sleep field has expanded to non- mammalian organisms, allowing the application of novel genetic approaches. We have developed an inducible-sleep system in the nematode C. elegans, and we are using the powerful molecular- genetic tools of this system to discover novel components of sleep regulation. We have found that forced expression of the C. elegans Epidermal Growth Factor (EGF) homolog LIN-3 induces a sleep-like state at any stage. By assaying for resistance to this effect we have identified several effectors of EGF-induced sleep, including the EGF receptor (LET-23/EGFR) and the ERG (ether-a- go-go related gene) homolog UNC-103, a voltage-gated potassium channel. We aim to determine the neurons in which UNC-103 activity is required for the inhibition of activity during sleep. We will examine potential changes in UNC-103 levels and localization in response to EGF expression, to make a connection between sleep signals and their downstream effects on neuronal excitability. Our studies are expected to shed light on mammalian sleep regulation, as EGF ligand administration in mammals has a conserved soporific effect. We will also investigate the timing of C. elegans sleep behavior, which occurs rhythmically with molting rather than with circadian periodicity. We will examine LIN-3 expression for oscillation with the molting cycle, to connect behavior with developmental timing cues. This analysis is relevant to the regulation of sleep by biological clocks in mammals, as the molting cycle is regulated by the C. elegans homolog of the circadian gene PER. Lastly, we will identify additional effectors of sleep regulation. We have found that EGFR activation within a single neuron, ALA, triggers EGF-induced sleep. We hypothesize that this neuron releases a signal that in turn triggers sleep across the neuronal network. We will identify this signal through RNAi-based functional genomics, and characterize its function using molecular-genetic approaches.

描述（由申请人提供）：睡眠不足会损害认知表现，而慢性睡眠丧失与心脏代谢疾病的风险增加有关，包括肥胖和高血压。尽管睡眠的重要性至关重要，但其功能仍然存在争议，并且执行其执行的分子​​机制才被阐明。睡眠被认为是由保守的遗传机制调节的，在过去的十年中，睡眠场已扩展到非哺乳动物生物，从而允许应用新颖的遗传方法。我们已经在线虫秀丽隐杆线虫中开发了一个诱导的睡眠系统，我们正在使用该系统的强大分子遗传工具来发现睡眠调节的新成分。我们发现，秀丽隐杆线虫表皮生长因子（EGF）同源物的强迫表达在任何阶段都会诱导睡眠状状态。通过测定对这种作用的抗性，我们已经确定了EGF诱导的睡眠的几个效应子，包括EGF受体（LET-23/EGFR）和ERG（Ether-A-Go-Go相关基因）同源性UNC-103，电压门控钾通道。我们旨在确定在睡眠期间抑制活动需要UNC-103活性的神经元。我们将检查UNC-103水平的潜在变化和响应EGF表达的定位，以在睡眠信号与其下游对神经元兴奋性的影响之间建立联系。我们的研究有望阐明哺乳动物的睡眠调节，因为哺乳动物中的EGF配体给药具有保守的可毒作用。我们还将研究秀丽隐杆线虫睡眠行为的时机，这是通过摩擦而不是昼夜节日时间进行节奏的。我们将检查LIN-3表达与摩擦周期的振荡，以将行为与发育时机提示联系起来。该分析与哺乳动物中生物钟对睡眠的调节有关，因为摩擦周期受到昼夜节律基因的秀丽隐杆线虫同源物的调节。最后，我们将确定睡眠调节的其他效果因子。我们发现，在单个神经元中，ALA的EGFR激活会触发EGF诱导的睡眠。我们假设该神经元释放出一个信号，该信号反过来触发了整个神经元网络的睡眠。我们将通过基于RNAi的功能基因组学识别该信号，并使用分子遗传学方法来表征其功能。