Analysis of EGFR-dependent sleep in C. elegans

线虫 EGFR 依赖性睡眠分析

基本信息

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
项目类别：
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

项目摘要

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）：睡眠障碍会损害认知能力，长期睡眠不足与肥胖和高血压等心脏代谢疾病的风险增加有关。尽管睡眠具有根本重要性，但其功能仍然存在争议，其执行的分子机制才刚刚开始被阐明。睡眠被认为是由保守的遗传机制调节的，在过去的十年中，睡眠领域已扩展到非哺乳动物生物体，从而允许应用新的遗传方法。我们在线虫中开发了一种诱导睡眠系统，并且我们正在利用该系统强大的分子遗传学工具来发现睡眠调节的新成分。我们发现，强制表达线虫表皮生长因子 (EGF) 同源物 LIN-3 会在任何阶段诱导类睡眠状态。通过分析对这种效应的抵抗力，我们已经确定了 EGF 诱导睡眠的几种效应子，包括 EGF 受体 (LET-23/EGFR) 和 ERG（ether-a-go-go 相关基因）同源物 UNC-103，一种电压-门控钾通道。我们的目标是确定哪些神经元需要 UNC-103 活性来抑制睡眠期间的活动。我们将检查 UNC-103 水平和定位响应 EGF 表达的潜在变化，以在睡眠信号及其对神经元兴奋性的下游影响之间建立联系。我们的研究有望揭示哺乳动物的睡眠调节，因为给予哺乳动物 EGF 配体具有保守的催眠作用。我们还将研究秀丽隐杆线虫睡眠行为的时间，该行为随着蜕皮而有节奏地发生，而不是按照昼夜节律周期性发生。我们将检查 LIN-3 表达随蜕皮周期的振荡，以将行为与发育时间线索联系起来。该分析与哺乳动物生物钟对睡眠的调节有关，因为蜕皮周期是由昼夜节律基因 PER 的线虫同源物调节的。最后，我们将确定睡眠调节的其他效应器。我们发现单个神经元 ALA 内的 EGFR 激活会触发 EGF 诱导的睡眠。我们假设该神经元释放信号，进而触发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表征其功能。