Brainstem circuitry for sleep-wake control

项目总结/摘要 现在很清楚，睡眠质量差会对健康造成严重影响，但睡眠药物仍然很重要。 需要安全有效的睡眠辅助。在了解大脑如何调节睡眠方面的最新重大进展- 唤醒周期开辟了新的研究路线，揭示了非快速眼的复杂调节网络 运动（NREM）睡眠控制，包括多个新的睡眠促进神经元群体。一系列 我们实验室最近的工作已经证明了面旁带（PZ）神经元在睡眠中的重要性 诱导和维持，并在此区域的GABA能神经元的促进深，恢复阶段 NREM睡眠称为慢波睡眠（SWS）。然而，在理解 PZ神经元控制睡眠的细胞和突触回路基础。长期目标是了解 PZ神经元促进睡眠的细胞和回路基础。核心假设是脑干 含有PZ GABA能神经元的亚群，其对于产生 SWS和皮层慢波活动（SWA）。拟议研究的基本原理是，了解如何 PZ促进睡眠是操纵这种促进睡眠回路的关键的第一步， 随后的转化研究集中在PZ上，旨在减少与睡眠中断相关的负担。 睡眠-觉醒障碍以及其他神经系统疾病。我们的假设将通过追踪两个 具体目的：1）揭示特异性促进睡眠的PZ GABA能亚群;和2）阐明 PZ GABA能神经元直接影响皮层活动的神经回路。以强为导 初步数据，在目标1中，我们将揭示PZ小清蛋白表达GABA能神经元在睡眠中的作用， 使用遗传驱动的损伤，化学遗传学/光遗传学激活/抑制， 在目标2中，我们将揭示一个直接的途径，通过该途径，PZ直接 影响丘脑-皮层活动并驱动SWS的SWA特征，使用体外电生理学， PZ GABA能投射到丘脑的光遗传学激活，PZ GABA能的体内Ca 2+成像 神经元投射到丘脑，和PZGABA在丘脑损伤小鼠的化学发生激活。的 这种方法在智力和技术上都是创新的，因为它代表了一种新的和实质性的基础， 了解睡眠调节，因为它采用了最先进的方法的新组合。的 拟议的研究是重要的，因为它预计将提供关键的知识，分子和细胞 调节睡眠的机制。最终，这些知识有望指导 更好地调节睡眠-觉醒行为和减轻负担的治疗和干预策略 与睡眠中断有关，不仅与睡眠障碍有关，而且与许多神经系统和 在美国和世界范围内，这些心理障碍代表了巨大的成本。

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 sleep promoting; and 2) elucidate the neuronal circuits by which PZ GABAergic 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 GABAergic projections to the thalamus, in vivo Ca2+ imaging of PZ GABAergic neurons projecting to the thalamus, and chemogenetic activation of PZGABA 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.