Medullary Circuitry Regulating Slow-Wave-Sleep

描述(申请人提供)：调节慢波睡眠(SWS)的皮质下结构与其脑电(EEG)的相关性尚不完全清楚。这种基本知识鸿沟的持续存在是一个重要的问题，因为它降低了我们调节或适当操纵大脑睡眠回路的能力，并阻碍了我们治疗和缓解睡眠中断引起的生理障碍的能力。我的长期目标是了解延髓副面区(PZ)中的神经元如何参与SWS和皮质慢波活动(SWA)的调节，后者与记忆巩固、突触稳态和皮质可塑性等基本神经生物学过程有关。在这一特定应用中的目标是确定)选择性激活GABA能PZ神经元是否促进自由行为动物的SWS和皮质SWA；2)GABA能PZ神经元如何在功能上、突触上与能够调制皮质EEG的电路连接；以及3)其他非GABA能PZ神经元可能如何参与SWS和皮质SWA的调节。中心假说是GABA能和非GABA能PZ神经元亚群由SWS促进神经元组成，它们通过向上投射到臂旁核和基底前脑产生SWS和皮质SWA。这项拟议研究的基本原理是，确定调节SWS和皮质SWA的相关PZ神经元是控制它们并减少睡眠障碍患者所经历的功能障碍的关键第一步。在强劲的初步数据指导下，将通过追求三个特定目标来验证这一假说：1)利用一种新开发的基因靶向技术，确定GABA能旁面区(PZ)神经元的急性和选择性激活是否能够在自由行为的动物中产生SWS和皮质SWA；2)结合基因靶向作图和光遗传学，确定GABA能PZ神经元在体内有效驱动SWS和SWA的突触基础；3)利用一种新的cre驱动小鼠系，以及类似于AIMS 1和2中使用的技术，确定非GABA能PZ神经元是否参与了SWS和皮质SWA的调节。这种方法在智力和技术上都是创新的，因为它代表了与当代睡眠调节电路模型的新的实质性背离，也因为它采用了新开发和验证的遗传驱动方法的新组合。这项拟议的研究意义重大，因为它有望纵向推进和扩大对大脑睡眠产生的细胞和突触机制以及PZ在这一调节中的作用的理解。归根结底，这些知识有可能为开发治疗方法提供信息，以减少美国越来越多的睡眠障碍患者经历的功能障碍和负面健康影响。

DESCRIPTION (provided by applicant): The sub-cortical structures regulating slow-wave-sleep (SWS) and its electroencephalogram (EEG) correlate are incompletely understood. Continued existence of this fundamental knowledge gap represents an important problem because it reduces our ability to modulate or appropriately manipulate the brain's sleep circuitry and hampers our ability to treat and alleviate the physiological disorders that result from sleep disruption. My long- term goal is to understand how neurons in the medullary parafacial zone (PZ) contribute to the regulation of SWS and cortical slow-wave-activity (SWA), the latter of which is linked to fundamental neurobiological processes like memory consolidation, synaptic homeostasis and cortical plasticity. The objectives in this particular application is to determine ) if selective activation of GABAergic PZ neurons promotes SWS and cortical SWA in freely behaving animals; 2) how GABAergic PZ neurons are functionally, synaptically connected with circuitry capable of modulating the cortical EEG; and 3) how other non-GABAergic PZ neurons might contribute to the regulation of SWS and cortical SWA. The central hypothesis is that subpopulations of GABAergic and non-GABAergic PZ neurons comprise a delimited node of SWS-promoting neurons, which generate SWS and cortical SWA through ascending projections to the parabrachial nucleus and basal forebrain. The rationale for the proposed research is that identifying the relevant PZ neurons that regulate SWS and cortical SWA represents a critical first step towards manipulating them and reducing the dysfunction experienced by individuals with sleep disorders. Guided by strong preliminary data, this hypothesis will be tested by pursuing three specific aims: 1) using a newly developed and genetically targeted technique, determine if acute and selective activation of GABAergic Parafacial Zone (PZ) neurons is capable of generating SWS and cortical SWA in freely behaving animals; 2) using a combination of genetically targeted mapping and optogenetics, determine the synaptic basis by which GABAergic PZ neurons potently drive SWS and SWA in vivo; 3) using a new cre-driver mouse line and similar techniques to those employed in Aims 1 and 2, determine if non-GABAergic PZ neurons contribute to the regulation of SWS and cortical SWA. The approach is intellectually and technically innovative because it represents a new and substantive departure from contemporary circuit models of sleep regulation and because it employs a novel combination of newly developed and validated genetically-driven approaches. The proposed research is significant, because it is expected to vertically advance and expand understanding of the cellular and synaptic mechanisms by which brain sleep is generated and the role of the PZ in this regulation. Ultimately, such knowledge has the potential to inform the development of treatments to reduce the dysfunction and negative health effects experienced by a growing number of patients with sleep disorders in the United States.