Genetic and Neuronal Mechanisms that Regulate Zebrafish Sleep

调节斑马鱼睡眠的遗传和神经机制

• 批准号: 10624762
• 负责人: David Aaron Prober
• 金额: $ 125.63万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2029-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10624762
• 关键词: Animals; Behavior; Behavioral Assay; Brain; Candidate Disease Gene; Cells; Circadian Rhythms; Complement; Coupled; Cues; Darkness; Economic Burden; Genes; Genetic; Health; Homeostasis; Human; Hypothalamic structure; Light; Mediating; Melatonin; Melatonin Receptors; Mental disorders; Monitor; Morbidity - disease rate; Mus; Neurons; Neuropeptides; Pathway interactions; Phenotype; Prevalence; Process; Research; Resolution; Rodent Model; Sleep; Sleep Disorders; Sleep Disorders Therapy; Testing; Time; Wakefulness; Zebrafish; candidate validation; circadian; circadian regulation; genome wide association study; genome-wide analysis; mutant; neuronal circuitry; novel; novel strategies; novel therapeutics; optogenetics; poor sleep; programs; sleep health; sleep regulation

ABSTRACT Sleep disorders are pervasive, contribute to morbidity in several psychiatric disorders, and cause an annual economic burden of $100 billion. However, despite its importance for health, the mechanisms that regulate sleep are poorly understood. We are taking a new approach to this problem by exploiting useful features of zebrafish to answer an important and basic question: What genetic and neuronal mechanisms regulate sleep? Sleep is regulated by a homeostatic process that reflects internal cues of sleep need and a circadian process that is entrained by environmental cues and restricts sleep to the appropriate time. Sleep is also directly and rapidly regulated by a phenomenon known as masking, in which light induces wake and dark induces sleep in diurnal animals. Factors that regulate the homeostatic process have been identified, including our recent finding that the serotonergic raphe promote sleep homeostasis in zebrafish and mice. We also showed that melatonin is essential for circadian regulation of sleep in zebrafish, and identified a pathway in the brain that regulates masking. Here we build upon these discoveries to elucidate mechanisms that underlie homeostatic, circadian, and light-dependent regulation of sleep. We will investigate these mechanisms using zebrafish, a diurnal vertebrate with several advantages that complement rodent models, using a combination of genetic, optogenetic, and chemogenetic perturbations coupled with high-throughput behavioral assays and whole-brain neuronal activity monitoring with single cell resolution. In Project 1, we will identify raphe subsystems that promote sleep homeostasis, and identify genetic and neuronal circuits that act upstream and downstream of these subsystems in sleep control. In Project 2, we will identify melatonin receptors that mediate the sleep-promoting function of melatonin, and also perform a screen to identify neurons through which melatonin implements circadian regulation of sleep. Project 3 builds on our recent discovery that the hypothalamic neuropeptide prokineticin 2 suppresses both light- and dark-induced masking behavior. Similar to Project 1, we will identify genetic and neuronal circuits that act upstream and downstream of prokineticin 2 to regulate masking. In Project 4, we will validate a large number of human sleep disorder candidate genes that were identified by genome-wide association studies. We will do so by leveraging zebrafish to efficiently and inexpensively generate and test many mutant lines for sleep phenotypes. We will determine the mechanisms through which validated candidate genes regulate sleep, and integrate these genes into the pathways identified in Projects 1-3. The homeostatic (Project 1), circadian (Project 2) and light-dependent (Project 3) mechanisms that regulate sleep, as well as the sleep disorder genes identified in humans and validated in zebrafish (Project 4), are likely to be integrated at multiple levels to produce either sleep or wakefulness. This research program provides a unified platform to explore interactions between genes and neurons identified in each project. This will allow us to derive a comprehensive understanding of mechanisms that regulate sleep, and will set the stage for novel therapies for sleep disorders.

摘要 睡眠障碍是普遍存在的，导致几种精神疾病的发病率， 1000亿美元的经济负担。然而，尽管睡眠对健康很重要， 我们对此知之甚少。我们正在采取一种新的方法来解决这个问题，即利用斑马鱼的有用特征 回答一个重要而基本的问题：是什么遗传和神经机制调节睡眠？睡眠是 受反映睡眠需要的内部线索的稳态过程和昼夜节律过程的调节， 受环境因素的影响，并将睡眠限制在适当的时间。睡眠也直接而迅速地 由一种称为掩蔽的现象调节，在这种现象中，白天光线引起觉醒，黑暗引起睡眠。 动物调节体内平衡过程的因素已经被确定，包括我们最近的发现， 神经元能中缝促进斑马鱼和小鼠的睡眠稳态。我们还发现褪黑激素 对斑马鱼睡眠的昼夜节律调节至关重要，并确定了大脑中调节睡眠的途径。 伪装在此，我们以这些发现为基础，阐明了体内平衡，昼夜节律， 和睡眠的光依赖调节。我们将研究这些机制，使用斑马鱼，一个昼夜 脊椎动物具有补充啮齿动物模型的几个优点，使用遗传，光遗传， 和化学遗传学扰动，再加上高通量行为测定和全脑神经元 具有单细胞分辨率的活动监测。在项目1中，我们将确定促进睡眠的中缝亚系统 稳态，并确定遗传和神经元回路，这些子系统的上游和下游的行为 睡眠控制。在项目2中，我们将确定褪黑激素受体介导的睡眠促进功能， 褪黑激素，还执行筛选以识别褪黑激素通过其实现昼夜节律的神经元 睡眠的调节。项目3建立在我们最近发现的下丘脑神经肽前动力素2的基础上， 抑制光和暗诱导的掩蔽行为。与项目1类似，我们将确定基因和 作用于前动力蛋白2上游和下游以调节掩蔽的神经元回路。在项目4中，我们 验证了大量的人类睡眠障碍候选基因，这些基因是通过全基因组鉴定的。 协会研究。我们将通过利用斑马鱼来有效和廉价地产生和测试许多 睡眠表型的突变株系。我们将确定验证候选基因的机制 调节睡眠，并将这些基因整合到项目1-3中确定的途径中。稳态（Homeostatic） 1），昼夜节律（项目2）和光依赖（项目3）调节睡眠的机制，以及睡眠 在人类中鉴定并在斑马鱼中验证的疾病基因（项目4），可能在多个基因组中整合。 产生睡眠或清醒的水平。本研究计划提供了一个统一的平台， 每个项目中确定的基因和神经元之间的相互作用。这将使我们能够得出一个全面的 了解调节睡眠的机制，并将为睡眠障碍的新疗法奠定基础。

项目成果

First steps into the cloud: Using Amazon data storage and computing with Python notebooks.

• DOI: 10.1371/journal.pone.0278316
• 发表时间: 2023
• 期刊: PloS one
• 影响因子: 3.7

The zebrafish mutant dreammist implicates sodium homeostasis in sleep regulation.

• DOI: 10.7554/elife.87521
• 发表时间: 2023-08-07
• 期刊: eLife
• 影响因子: 7.7
• 作者: Barlow IL;Mackay E;Wheater E;Goel A;Lim S;Zimmerman S;Woods I;Prober DA;Rihel J
• 通讯作者: Rihel J

Validation of Candidate Sleep Disorder Risk Genes Using Zebrafish.

• DOI: 10.3389/fnmol.2022.873520
• 发表时间: 2022
• 期刊: FRONTIERS IN MOLECULAR NEUROSCIENCE
• 影响因子: 4.8
• 作者: Tran, Steven;Prober, David A.
• 通讯作者: Prober, David A.