Neuropeptide S and arousal

神经肽 S 和唤醒

• 批准号: 10390209
• 负责人: Luis De Lecea
• 金额: $ 62.54万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-09 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10390209
• 关键词: Acetylcholine; Acute; Address; Affect; Amygdaloid structure; Anatomy; Anxiety; Anxiety Disorders; Architecture; Area; Arousal; Behavior; Behavioral; Body Temperature; Brain; Brain region; Breathing; Cell Nucleus; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Data; Development; Dopamine; Enterobacteria phage P1 Cre recombinase; Etiology; Fiber; Gene Mutation; Genetic study; Human; Individual; Injections; Internal Ribosome Entry Site; Interneurons; Knock-in Mouse; Knockout Mice; Label; Lead; Ligands; Light; LoxP-flanked allele; Maps; Medial; Mediating; Messenger RNA; Methods; Motor Activity; Mus; Mutation; Names; Narcolepsy; Neurons; Neuropeptide Gene; Neuropeptide Receptor; Neuropeptides; Neurotensin; Norepinephrine; Pattern; Peptides; Photometry; Physiological; Pontine structure; Population; Process; Regulation; Reporter; Role; Serine; Signal Transduction; Sleep; Sleep Architecture; Sleep Disorders; Sleep Wake Cycle; Stress; Structure; Subgroup; System; Testing; Thalamic structure; Viral; Wakefulness; base; brain cell; combinatorial; emotional stimulus; experimental study; feeding; gain of function mutation; hypocretin; improved; knock-down; neuropsychiatric disorder; neurotransmission; neurotransmitter release; non rapid eye movement; novel therapeutics; optogenetics; promoter; response; sleep regulation; tool; vigilance

Abstract Neuropeptides have a critical role in modulating sleep and wakefulness and offer unique opportunities to treat sleep disorders. Among them, Neuropeptide S shows outstanding features: i) Administration of NPS increases wakefulness and reduces anxiety; ii) Neuropeptide S knockout mice show display increased NREM and anxiety; iii) Mutations of the NPS receptor that give rise to overactive signaling result in short sleep in humans and mice; iv) Expression of NPS is restricted to a few thousand neurons distributed across five main clusters in the basomedial amygdala, dorsomedial thalamus, Kolliker-Fuse/parabrachial area, pericoerulear region and nucleus incertus. These regions have been directly or indirectly associated with arousal and anxiety, but the detailed mechanisms as to how the modulate sleep architecture are unknown. We have recently generated a new line of mice expressing cre recombinase under the control of the endogenous NPS gene promoter (NPS-IRES-cre mice). Here we propose to use these mice and a combination of circuit mapping tools to decipher the mechanisms by which NPS modulates sleep/wake cycle. First, we will use viral-mediated tracing to determine if the five clusters of NPS+ neurons are interconnected, and their anatomical relationship with known arousal circuits. In a second aim, we will use fiber photometry to determine the activity profiles of the five clusters of NPS cells across the sleep/wake cycle and in response to stress and positive emotional stimuli. We will also determine which arousal circuits are activated by optogenetic stimulation of NPS, and which circuits activate NPS neurons. We will also assess whether NPS stimulation affects locomotor activity, anxiety, core body temperature and other physiological variables that may confound the arousal effect. In aim 3, we will test whether individual clusters of NPS neurons are necessary for NPS’s effects on sleep by using opto and chemogenetic inhibition. Finally, we will use a CRISPR-based approach to introduce NPS gene mutations in individual NPS+ cell clusters and determine whether NPS release in these brain regions is essential to control sleep/wake architecture and anxiety behaviors. The results from these experiments will shed new light into the function of NPS and NPS+ neurons, as well as the interconnection between them and arousal circuits. These data may lead to improved treatments of neuropsychiatric disorders associated with imbalances in arousal systems.

抽象的 神经肽在调节睡眠和觉醒方面发挥着关键作用，并提供独特的治疗机会 睡眠障碍。其中，Neuropeptide S表现出突出的特点： i) NPS给药量增加 保持清醒并减少焦虑； ii) 神经肽 S 敲除小鼠表现出 NREM 和焦虑增加； iii) NPS 受体突变导致信号过度活跃，导致人类和小鼠睡眠不足； iv) NPS 的表达仅限于分布在大脑中五个主要簇中的数千个神经元。 基底内侧杏仁核、背内侧丘脑、Kolliker-Fuse/臂旁区、蓝周区和核 犹豫不决。这些区域与唤醒和焦虑直接或间接相关，但详细的 关于如何调节睡眠结构的机制尚不清楚。我们最近生成了一条新线 在内源性 NPS 基因启动子 (NPS-IRES-cre) 控制下表达 cre 重组酶的小鼠 老鼠）。在这里，我们建议使用这些鼠标和电路映射工具的组合来破译 NPS 调节睡眠/觉醒周期的机制。首先，我们将使用病毒介导的追踪来确定是否 五个 NPS+ 神经元簇是相互连接的，它们的解剖学关系与已知的唤醒 电路。第二个目标是，我们将使用光纤光度测定法来确定五个簇的活性概况。 NPS 细胞贯穿睡眠/觉醒周期并对压力和积极的情绪刺激做出反应。我们还将 确定哪些唤醒回路被 NPS 的光遗传学刺激激活，以及哪些回路激活 NPS 神经元。我们还将评估 NPS 刺激是否影响运动活动、焦虑、核心身体 温度和其他可能混淆唤醒效果的生理变量。在目标 3 中，我们将测试 通过使用 Opto 和 NPS 对睡眠的影响，单个 NPS 神经元簇是否是必要的 化学遗传学抑制。最后，我们将使用基于 CRISPR 的方法将 NPS 基因突变引入 单个 NPS+ 细胞簇，并确定这些大脑区域中 NPS 的释放是否对于控制至关重要 睡眠/觉醒结构和焦虑行为。 这些实验的结果将为 NPS 和 NPS+ 神经元的功能以及 它们和唤醒电路之间的互连。这些数据可能会导致治疗方法的改进 与唤醒系统失衡相关的神经精神疾病。