Genetic Analysis of Sleep Disorders in Zebrafish

描述(由申请人提供)： 超过10%的人患有慢性睡眠障碍，但调节睡眠的遗传机制在很大程度上是未知的。发现有缺陷的下丘脑素/食欲素(Hcrt)信号是发作性睡病的原因，这为睡眠研究提供了一个遗传学切入点，但尚未找到治疗这种疾病的有效方法。此外，只有一小部分睡眠障碍与嗜睡症有关，这表明控制睡眠和清醒的其他基因仍有待确定。这项建议的目的是利用斑马鱼作为一种简单且经济有效的脊椎动物模型系统来研究Hcrt信号和睡眠的遗传学。斑马鱼是这些研究的一个极好的模型，因为它们易于接受高通量的基因筛选，而且与无脊椎动物不同，它们拥有Hcrt同源基因和调节哺乳动物睡眠的基本大脑结构。 我们已经证明，Hcrt的过度表达会导致斑马鱼幼体出现类似失眠的表型。在具体目标1中，我们将使用药物和基因突变来确定Hcrt过表达诱导这种表型的遗传和神经学机制。这些实验将提高对Hcrt功能的理解，并可能为慢性失眠的基础提供线索。在具体目标2中，我们将使用斑马鱼幼体进行高通量遗传筛选，以确定新的睡眠调节剂。这一筛选针对的是可能与睡眠有关的基因类别，并为治疗睡眠障碍提供了良好的药物靶点。 我目前的职业目标是在两年内完成我的博士后研究，并在顶尖大学或研究机构开设自己的实验室，成为一名独立研究员。我的长期职业目标是阐明调节睡眠/清醒状态的遗传和神经学机制，希望这一知识将导致睡眠障碍的治疗。除了我的导师，我还组建了一个专家顾问团队，他们将帮助我进行培训，帮助我成为一名成功的独立调查员。 相关性：超过10%的人患有睡眠障碍，但人们对睡眠障碍的原因或睡眠是如何调节的知之甚少。这项提议将以斑马鱼为模型系统，通过研究一种导致人类睡眠障碍发作性睡病的基因来研究睡眠是如何受到调节的。斑马鱼还将被用来识别调节睡眠的新基因，并可能参与人类睡眠障碍。

DESCRIPTION (provided by applicant): Greater than 10% of humans suffer chronic sleep disorders, but the genetic mechanisms that regulate sleep are largely unknown. The identification of defective Hypocretin/Orexin (Hcrt) signaling as a cause of narcolepsy provided a genetic entry point into sleep research, but an effective treatment for this disorder has not yet been found. Moreover, only a small fraction of sleep disorders are associated with narcolepsy, indicating that additional genes that control sleep and wakefulness remain to be identified. The objective of this proposal is to use zebrafish as a simple and cost-effective vertebrate model system to study the genetics of Hcrt signaling and sleep. Zebrafish are an excellent model for these studies because they are amenable to high-throughput genetic screens and,unlike invertebrates, have a Hcrt ortholog and the basic brain structures that regulate sleep in mammals. We have shown that Hcrt overexpression causes an insomnia-like phenotype in zebrafish larvae. In Specific Aim 1, we will use pharmacological agents and genetic mutants to determine the genetic and neurologic mechanisms by which Hcrt overexpression induces this phenotype. These experiments will improve understanding of Hcrt function and may provide clues to the basis of chronic insomnia. In Specific Aim 2, we will use zebrafish larvae to perform a high-throughput genetic screen to identify novel sleep regulators. This screen is targeted to classes of genes that are likely to be involved in sleep and provide good drug targets to treat sleep disorders. My immediate career goal is to complete my postdoctoral studies within 2 years and to start my own lab as an independent investigator at a top university or research institute. My long-term career goal is to elucidate the genetic and neurologic mechanisms that regulate sleep/wake states, with the hope that this knowledge will lead to treatments for sleep disorders. In addition to my mentor, I have assembled a team of expert consultants who will aid in my training to help me become a successful independent investigator. Relevance: Over 10% of humans suffer from sleep disorders, yet little is known about the causes of sleep disorders or how sleep is regulated. This proposal will use zebrafish as a model system to study how sleep is regulated by studying a gene whose loss causes the human sleep disorder narcolepsy. Zebrafish will also be used to identify new genes that regulate sleep and may be involved in human sleep disorders.