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Purinergic Mechanisms in Homeostatic Sleep Control

稳态睡眠控制中的嘌呤能机制

基本信息

批准号：
10215231
负责人：
RADHIKA BASHEER
金额：
--
依托单位：
VA BOSTON HEALTH CARE SYSTEM
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-10-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10215231
关键词：
Acetylcholine Acute Adenosine Adenosine Triphosphate Affect Attenuated Bilateral Biological Assay Biology Bioluminescence Brain Brain region Chronic Coupled Data Drowsiness Electroencephalography Electrophysiology (science)Feedback Frequencies Genotype Glutamate Receptor Glutamates Goals Grant Health High Pressure Liquid Chromatography Homeostasis In Vitro Lesion Light Liquid Chromatography Measures Mecamylamine Mediating Mental Depression Mental Health Methodology Microdialysis Military Personnel Mus Neurons Neurotransmitter Receptor Neurotransmitters Nicotinic Antagonists Nicotinic Receptors Optics Parvalbumins Performance Play Post-Traumatic Stress Disorders Presynaptic Receptors Presynaptic Terminals Proton Pump Receptor Activation Recovery Reporting Risk Risk Factors Role Sleep Sleep Deprivation Sleep Disorders Sleep disturbances Sleeplessness Slice Soldier Source System Testing Therapeutic Intervention Time Veterans War active duty adeno-associated viral vector basal forebrain cell type cholinergic cholinergic neuron erythroidine experience experimental study extracellular in vivo insight military veteran neurochemistry non rapid eye movement novel optogenetics physical conditioning postsynaptic presynaptic receptor response sleep regulation tandem mass spectrometry therapeutic development vector

项目摘要

The broad objective of this proposal is to understand the cellular mechanisms which control sleepiness. Sleep deprivation (SD) and insomnia are experienced by ~40% of military personnel immediately after deployment and SD is a well-known risk factor that adversely affects the mental health of veterans. The basal forebrain (BF) is an important brain region for controlling the increased propensity for sleep after SD i.e. sleep homeostasis. Previous grant cycles investigated how sleep homeostasis is mediated by inhibition of BF wake-promoting neurons via increases in the extracellular concentration of adenosine (ADex), and investigated the role of extracellular adenosine triphosphate (ATPex) as a source of ADex. However, the circuit and neurotransmitter basis for these increases in ATPex and ADex has remained unresolved. At least three types of cortically-projecting, wake-active neurons coexist within BF: cholinergic (ChAT+), parvalbumin expressing GABAergic (PV+) and glutamatergic (vGluT2+). Selective lesion studies of BF ChAT+ neurons suggested that they are required for SD-induced increases in ADex and for sleep homeostasis. However, irreversible lesions may cause degenerative changes which go beyond loss of ChAT+ neurons. Thus, it is important to test the role of ChAT+ neurons using reversible stimulation or inactivation experiments. Furthermore, the need for intact ChAT+ neurons could reflect either ATPex/ADex release from ChAT+ neurons themselves and/or ChAT+ modulation of neighboring, non-cholinergic neurons. Thus, in this application we will use our novel methodology of optodialysis (Zant et al., 2016) to combine reversible, neuronal-subtype-specific optogenetic manipulations with simultaneous in vivo microdialysis to measure neurochemical changes in the presence and absence of selective antagonists for particular neurotransmitter receptors to delineate functional interactions between neuronal subtypes. Recent reports showed that optogenetic stimulation of vGluT2+ neurons excited ChAT+ neurons whereas PV+ neurons had only a weak influence on ChAT+ neurons. Furthermore, BF glutamate receptor activation enhanced acetylcholine (ACh) release in cortex and increased ADex levels in BF. These findings provide the basis for our hypothesis that local positive feedback between BF ChAT+ and vGluT2+ but not PV+ neurons leads to the ATPex/ADex increases which underlie the homeostatic sleep response. Three specific aims (SA) are proposed towards this goal: We will examine the role of cholinergic, parvalbumin expressing GABAergic, and glutamatergic neurons in sleep homeostasis using optogenetic stimulation and inhibition (SA1) and BF increases of ATPex and ADex using optodialysis (SA2). Using in vitro electrophysiology and in vivo optodialysis, SA3 will examine the hypothesis that a local positive feedback between cholinergic and glutamatergic neurons leads to BF ATPex/ADex increases during SD. Successful completion of these studies will provide insight into the cellular and neurochemical mechanisms underlying the homeostatic sleep response. Thereby, facilitating the development of therapeutic measures to reduce the deleterious effects of sleep loss in military personnel, veterans and people with sleep disorders.

该提案的主要目标是了解控制睡意的细胞机制。睡觉约 40% 的军事人员在部署后立即经历了剥夺 (SD) 和失眠 SD 是一个众所周知的风险因素，会对退伍军人的心理健康产生不利影响。基底前脑 (BF) 是 SD 后控制睡眠倾向增加的重要大脑区域，即睡眠稳态。之前的资助周期研究了如何通过抑制 BF 唤醒来介导睡眠稳态神经元通过增加细胞外腺苷 (ADex) 浓度，并研究了细胞外三磷酸腺苷 (ATPex) 作为 ADex 的来源。然而，电路和神经递质基础 ATPex 和 ADex 增加的问题仍未得到解决。 BF 内至少共存三种类型的皮质投射、唤醒活动神经元：胆碱能 (ChAT+)、小清蛋白表达 GABA 能 (PV+) 和谷氨酸能 (vGluT2+)。 BF ChAT+ 的选择性病变研究神经元表明它们是 SD 诱导的 ADex 增加和睡眠稳态所必需的。然而，不可逆的损伤可能会导致退行性变化，其程度超出了 ChAT+ 神经元损失的范围。因此，它是使用可逆刺激或失活实验来测试 ChAT+ 神经元的作用非常重要。此外，对完整 ChAT+ 神经元的需求可以反映 ChAT+ 神经元本身释放 ATPex/ADex 和/或邻近非胆碱能神经元的 ChAT+ 调节。因此，在这个应用程序中，我们将使用我们的小说光透析方法（Zant et al., 2016）结合可逆的、神经元亚型特异性的光遗传学同时体内微透析的操作来测量神经化学变化的存在和缺乏特定神经递质受体的选择性拮抗剂来描述功能相互作用神经元亚型之间。最近的报告表明，光遗传学刺激 vGluT2+ 神经元会兴奋 ChAT+神经元而PV+神经元对ChAT+神经元只有微弱的影响。此外，BF 谷氨酸受体激活增强了皮质中乙酰胆碱 (ACh) 的释放，并增加了 BF 中的 ADex 水平。这些发现为我们的假设提供了基础，即 BF ChAT+ 和 vGluT2+ 之间的局部正反馈但 PV+ 神经元不会导致 ATPex/ADex 增加，这是稳态睡眠反应的基础。三为实现这一目标提出了具体目标（SA）：我们将研究胆碱能、小清蛋白的作用使用光遗传学刺激在睡眠稳态中表达 GABA 能和谷氨酸能神经元使用光透析 (SA2) 抑制 (SA1) 和 BF 增加 ATPex 和 ADex。使用体外电生理学和体内光透析，SA3 将检验胆碱能和光之间局部正反馈的假设谷氨酸能神经元导致 SD 期间 BF ATPex/ADex 增加。成功完成这些研究将深入了解稳态睡眠反应背后的细胞和神经化学机制。因此，促进治疗措施的开发，以减少睡眠不足的有害影响军人、退伍军人和患有睡眠障碍的人。