Neural control of NREM sleep in the medulla

延髓 NREM 睡眠的神经控制

• 批准号: 10567029
• 负责人: Yueqing Peng
• 金额: $ 41.13万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-01 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10567029
• 关键词: Adult; Animals; Area; Arousal; Biological Phenomena; Brain; Brain region; Calcium; Data; Disease; Electrophysiology (science); FOS gene; Genetic; Genetic Markers; Glutamates; Human; Image; Immunochemistry; Investigation; Life; Maintenance; Maps; Modeling; Mus; Neurons; Organism; Output; Pathway interactions; Physiological; Physiology; Pilot Projects; Population; Preoptic Areas; Prevalence; Process; Research; Research Project Grants; Research Proposals; Role; Sleep; Sleep Disorders; Sleeplessness; Structure; System; Techniques; Testing; Transgenic Mice; United States National Institutes of Health; Viral; Wakefulness; Work; awake; cell type; circadian; experimental study; falls; follow-up; gain of function; in vivo; in vivo calcium imaging; innovation; loss of function; mouse genetics; neural; neural circuit; neuronal circuitry; neuroregulation; non rapid eye movement; novel; optogenetics; single-cell RNA sequencing; sleep behavior; sleep regulation; tool; transcriptome sequencing

Project Summary/Abstract In this proposal, we make use of optogenetic and chemogenetic tools, in vivo calcium imaging, RNA-seq, viral-based circuit tracing, and genetic targeting techniques to dissect the neural circuits that control sleep behavior in the mammalian brain. At a fundamental level, the work presented in this research proposal may provide valuable information for developing new treatments for various human sleep disorders, such as insomnia. Decades of studies have revealed neuromodulatory circuits as key regulators of sleep and wakefulness. However, most of these studies focus on the arousal system. Sleep-promoting neurons are theoretically required for the initiation and maintenance of sleep states based on the flip-flop model of wake-sleep switching, in which sleep-promoting neurons are mutually inhibitory with wake-promoting neurons. Previous studies have identified several brain structures that promote non-rapid eye movement (or NREM) sleep, but whether these structures are involved in the initiation or the maintenance remains largely unknown. In this research project, we will primarily focus on neural control of sleep initiation, i.e. the transition from wakefulness to sleep – under physiological conditions, always to NREM sleep. We aim to decipher the circuit mechanisms underlying the wake-sleep transition. In pilot studies, we have identified a novel population of glutamatergic neurons in the ventrolateral medulla (VLM) that project to the preoptic area (POA), a prominent sleep center. We present preliminary data showing that these VLM neurons are activated during wake-sleep transitions and optogenetic activation of these neurons induces long-lasting NREM sleep in awake mice. These results lead to our working hypothesis: VLM glutamatergic neurons induce the transition from wakefulness to NREM sleep, which subsequently activate their downstream targets to maintain NREM sleep. In Aim 1, we will activate and inactivate VLM glutamatergic neurons and examine the sufficiency and necessity of their activity in the wake-sleep transition. In Aim2, we will perform microendoscopic calcium imaging in VLM glutamatergic neurons and examine their activity during sleep. Then, we will use single-cell RNA-seq to identify genetic markers of sleep-active VLM glutamatergic neurons for further in vivo experiments. In Aim 3, we will use viral-based circuit tracing to characterize the inputs and outputs of VLM neurons and examine their function in sleep regulation. Together, the results obtained from this proposal will expand our understanding of the neural basis of sleep behavior.

项目摘要/摘要 在此提案中，我们利用光学遗传学和化学遗传学工具在体内钙 成像，RNA-seq，基于病毒的电路跟踪和遗传靶向技术以剖析 控制哺乳动物大脑睡眠行为的神经回路。在基本层面上 本研究建议中介绍的工作可能会为开发提供有价值的信息 针对各种人类睡眠障碍的新疗法，例如失眠。 数十年的研究揭示了神经调节性循环是睡眠的关键调节因子和 清醒。但是，大多数研究都集中在唤醒系统上。促进睡眠 从理论上讲，神经元是基于睡眠状态的主动和维护所必需的 尾流切换的触发器模型，其中促进睡眠神经元是相互的 抑制唤醒神经元。先前的研究已经确定了几个大脑 促进非比型眼运动（或NREM）睡眠的结构，但是这些是否是否 该计划涉及结构，或者维护仍然在很大程度上未知。在这个 研究项目，我们将主要关注睡眠计划的神经控制，即过渡 从觉醒到睡眠 - 在生理条件下，总是到NREM睡眠。我们 旨在破译唤醒睡眠过渡的电路机制。 在试点研究中，我们已经确定了新的谷氨酸神经元种群 腹侧延髓（VLM）该项目前往型前区域（POA），这是一个突出的睡眠 中心。我们提供了初步数据，表明这些VLM神经元在 这些神经元的唤醒性转变和光遗传激活会导致长期持续 NREM在清醒老鼠中睡觉。这些结果导致了我们的工作假设：VLM 谷氨酸能神经元诱导从清醒到NREM睡眠的过渡， 随后，激活其下游目标以保持NREM睡眠。在AIM 1中，我们将 激活和灭活VLM谷氨酸能神经元并检查足够和必要性 他们在尾流过渡中的活动。在AIM2中，我们将执行微镜钙 在VLM谷氨酸能神经元中进行成像，并检查其在睡眠期间的活性。然后，我们会的 使用单细胞RNA-seq鉴定睡眠活性VLM谷氨酸能的遗传标记 用于进一步体内实验的神经元。在AIM 3中，我们将使用基于病毒的电路跟踪到 表征VLM神经元的输入和输出并检查其在睡眠中的功能 规定。从本提案中获得的结果一起将扩大我们对 睡眠行为的神经元基础。