Neuropeptide S and arousal

摘要 神经肽在调节睡眠和清醒方面起着关键作用，并提供了独特的治疗机会 睡眠障碍。其中，神经肽S表现出突出的特点：1)给药增加 II)神经肽S基因敲除小鼠表现出NREM增加和焦虑； Iii)NPS受体的突变导致信号过度活跃，导致人类和小鼠睡眠时间过短； IV)NPS的表达仅限于分布在五个主要簇中的数千个神经元 杏仁基底内侧核、丘脑背内侧核、Kolliker-Fuse/臂旁区、髓周区和核团 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尚不清楚。我们最近生产了一条新的生产线 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跟踪来确定 NPS+神经元的五个簇是相互联系的，它们与已知的觉醒之间的解剖关系 电路。在第二个目标中，我们将使用纤维光度法来确定五个星团的活性分布 NPS细胞贯穿睡眠/清醒周期，对压力和积极的情绪刺激做出反应。我们还将 确定NPS的光遗传刺激激活了哪些唤醒电路，以及激活了哪些电路 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 温度和其他可能扰乱唤醒效果的生理变量。在目标3中，我们将测试 使用OPTO和AND研究NPS对睡眠的影响是否需要单独的NPS神经元簇 化学发生抑制。最后，我们将使用基于CRISPR的方法将NPS基因突变引入 并确定NPS在这些脑区的释放是否对控制 睡眠/觉醒结构和焦虑行为。 这些实验的结果将为了解NPS和NPS+神经元的功能以及 它们和唤醒回路之间的相互联系。这些数据可能会导致改善对 与唤醒系统失衡相关的神经精神障碍。

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.