Brainstem circuitry for sleep-wake control

项目摘要/摘要 现在很清楚，糟糕的睡眠质量会对健康产生巨大的影响，但安眠药仍然很流行 需要安全有效的睡眠辅助设备。最近在了解大脑如何调节睡眠方面取得了重大进展- 觉醒周期开启了新的调查路线，揭示了非快眼的复杂监管网络 运动(NREM)睡眠控制，包括多个新的促进睡眠的神经元群体。一系列 我们实验室最近的工作证明了副面区(PZ)神经元在睡眠中的关键重要性 促进深层恢复期GABA能神经元的诱导和维持 NREM睡眠称为慢波睡眠(SWS)。然而，在理解这一问题上存在着根本的差距 PZ神经元控制睡眠的细胞和突触电路基础。我们的长期目标是理解 PZ神经元促进睡眠的细胞和电路基础。中心假说是脑干 包含PZ GABA能神经元的一个亚群，这些神经元对于产生 SWS和皮层慢波活动(SWA)。这项拟议研究的基本原理是理解如何 PZ促进睡眠是操纵这一促进睡眠的回路的关键第一步，并将导致 随后的翻译研究以PZ为中心，旨在减轻与睡眠中断相关的负担 有睡眠-觉醒障碍，但也有其他神经障碍。我们的假设将通过追求两个 具体目标：1)发现一个特定于SWS促进的PZ GABA能亚群；2)阐明 PZ SWS促进神经元的神经元回路直接影响皮层活动。以Strong为指导 初步数据，在目标1中，我们将揭示PZ小白蛋白表达GABA能神经元在睡眠中的作用- 使用遗传驱动损伤、化学发生/光发生激活/抑制、 纤维光度测量和神经元追踪；在目标2中，我们将发现PZ直接通过 影响丘脑皮质活动并驱动SWS的SWA特性，利用体外电生理学， PZ投射到丘脑的光发生激活，投射到丘脑的PZ神经元的体内钙成像 丘脑，以及表达GABA能的PZ小白蛋白在丘脑损伤小鼠中的化学激活。这个 方法在智力和技术上都是创新的，因为它代表了 了解睡眠调节，因为它采用了一种最先进的方法的新组合。这个 拟议的研究具有重要意义，因为它有望提供分子和细胞的关键知识。 调节睡眠的机制。最终，这些知识有望指导 治疗和干预策略，以更好地调节睡眠唤醒行为和减轻负担 与睡眠障碍有关，不仅与睡眠障碍有关，而且还与许多神经和 心理障碍在美国和世界范围内代表着巨大的代价。

PROJECT SUMMARY/ABSTRACT It is now clear that poor sleep quality has dramatic health consequences, yet sleep medicine is still in great need for safe and efficient sleep aids. Recent major advances in understanding how the brain regulates sleep- wake cycles have opened new lines of investigations, revealing a complex regulatory network for non-rapid eye movement (NREM) sleep control that includes multiple new sleep-promoting neuronal populations. A series of recent work by our laboratory have demonstrated the critical importance of parafacial zone (PZ) neurons in sleep induction and maintenance, and of the GABAergic neurons in this region in promoting the deep, restorative stage of NREM sleep known as slow-wave sleep (SWS). There is a fundamental gap, however, in understanding the cellular and synaptic circuit basis by which PZ neurons control sleep. The long-term goal is to understand the cellular and circuit bases by which PZ neurons promote sleep. The central hypothesis is that the brainstem contains a sub-population of PZ GABAergic neurons that are both sufficient and necessary for the generation of SWS and cortical slow-wave activity (SWA). The rationale for the proposed research is that understanding how the PZ promotes sleep is a critical first step towards manipulating this sleep-promoting circuit and will lead to subsequent translational studies centered on the PZ aimed at reducing the burden of sleep disruption associated with sleep-wake disorders but also other neurologic disorders. Our hypothesis will be tested by pursuing two specific aims: 1) uncover a PZ GABAergic sub-population that is specifically SWS promoting; and 2) elucidate the neuronal circuits by which PZ SWS promoting neurons directly influence cortical activity. Guided by strong preliminary data, in aim 1, we will uncover the role of PZ Parvalbumin expressing GABAergic neurons in sleep- wake control using a combination of genetically-driven lesions, chemogenetic/optogenetic activation/inhibition, fiber photometry and neuronal tracing; and in Aim 2, we will uncover a direct pathway by which the PZ directly affects thalamo-cortical activity and drives the SWA characteristic of SWS, using in vitro electrophysiology, optogenetic activation of PZ projections to the thalamus, in vivo Ca2+ imaging of PZ neurons projecting to the thalamus, and chemogenetic activation of PZ Parvalbumin expressing GABAergic in thalamic lesioned mice. The approach is intellectually and technically innovative because it represents a new and substantive substrate of understanding sleep regulation and because it employs a novel combination of state-of-the-art approaches. The proposed research is significant because it is expected to provide critical knowledge of the molecular and cellular mechanisms by which sleep is regulated. Ultimately, such knowledge is expected to guide the development of therapeutic and interventional strategies to better regulate sleep-wake behavior and to reduce the burden associated with sleep disruption, not only associated with sleep disorders but also with many neurological and psychological disorders that represent a tremendous cost in the United States and worldwide.