Neuropeptide S and arousal

摘要 神经肽在调节睡眠和觉醒方面具有关键作用，并提供了独特的治疗机会。 睡眠障碍其中，神经肽S显示出突出的特点：i）施用抗氧化剂增加 ii）神经肽S敲除小鼠显示NREM和焦虑增加; iii）导致信号过度活跃的α受体突变导致人类和小鼠睡眠时间短; iv）在神经元中，E2的表达限于分布在五个主要簇中的几千个神经元。 杏仁核基底内侧区、丘脑背内侧区、Kolliker-Bronze/臂旁区、红核周围区和核 incertus。这些区域直接或间接地与唤醒和焦虑有关，但详细的 关于如何调节睡眠结构的机制是未知的。我们最近生产了一条新的生产线 表达cre重组酶的小鼠在内源性cre基因启动子的控制下（NPS-IRES-cre 小鼠）。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 睡眠调节睡眠/觉醒周期的机制。首先，我们将使用病毒介导的跟踪来确定 这五个神经元簇是相互连接的，它们与已知的唤醒的解剖关系 电路.在第二个目标中，我们将使用纤维光度法来确定五个簇的活性分布， 在整个睡眠/觉醒周期以及对压力和积极情绪刺激的反应中，我们还将 确定哪些唤醒回路被光遗传学刺激激活，哪些回路激活 神经元。我们还将评估神经刺激是否会影响运动活动、焦虑、核心体 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， 化学发生抑制最后，我们将使用一种基于CRISPR的方法，在 单个的β +细胞簇，并确定这些大脑区域中的β +释放是否对控制 睡眠/觉醒结构和焦虑行为。 这些实验的结果将为神经元和神经元的功能提供新的线索， 它们和唤醒回路之间的联系这些数据可能会导致改善治疗 与唤醒系统不平衡有关的神经精神障碍。

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.