Synaptic Basis of Sleep Cycle Control

睡眠周期控制的突触基础

• 批准号: 8006426
• 负责人: Robert W McCarley
• 金额: $ 35.4万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-09-30 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8006426
• 关键词: Accidents; Address; Adenosine; Adenosine A1 Receptor; Affect; Animal Model; Animal Testing; Animals; Antibodies; Area; Astrocytes; Attention; Autoradiography; Behavior; Behavioral; Behavioral Mechanisms; Binding; Biological; Brain; Brain Stem; Brain region; Cells; Cerebral cortex; Choline O-Acetyltransferase; Circadian Rhythms; Data; Deet; Dependence; Depressed mood; Detection; Disease; Dose; Dyes; Electroencephalography; Emergency Situation; Evaluation; Evolution; Feedback; Fluorescence; Foundations; Glial Fibrillary Acidic Protein; Glutamate Decarboxylase; Glutamates; Grant; Green Fluorescent Proteins; Health; Homeostasis; Human; Immunohistochemistry; In Vitro; Infusion procedures; Injection of therapeutic agent; Inosine; Label; Length; Life; Link; Measurement; Measures; Mediating; Mediator of activation protein; Medical; Membrane; Mental Depression; Messenger RNA; Methods; Microdialysis; Microglia; Microspheres; Modeling; Molecular; Motivation; Mus; NG-Nitroarginine Methyl Ester; National Institute of Mental Health; Neuroglia; Neurons; Nitric Oxide; Nitric Oxide Donors; Outcome; Pathway interactions; Performance; Perfusion; Pharmacology; Physiological; Potassium Channel; Prefrontal Cortex; Production; Property; Proteins; PubMed; Quality of life; Rattus; Receptor Activation; Response Latencies; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Schedule; Schools; Shapes; Sleep; Sleep Deprivation; Sleep Disorders; Sleep Wake Cycle; Sleeplessness; Slice; Soluble Guanylate Cyclase; Somatosensory Cortex; Source; Spatial Distribution; Specificity; Staining method; Stains; Stimulus; Synapses; System; Technology; Testing; Theophylline; Time; Up-Regulation; Wakefulness; Western Blotting; Whole-Cell Recordings; Work; adenosine deaminase; analog; awake; basal forebrain; basal forebrain cholinergic neurons; base; behavior measurement; behavior test; calbindin; calretinin; cell type; cholinergic; cholinergic neuron; cingulate cortex; design; extracellular; gamma-Aminobutyric Acid; human NOS2A protein; in vivo; inhibitor/antagonist; juvenile animal; mimicry; neurochemistry; neurophysiology; novel; postsynaptic; presynaptic; receptor; receptor binding; receptor density; research study; response; retrograde transport; selective expression; shift work; social; vigilance

DESCRIPTION (provided by applicant): Why do we get sleepy after being awake? Why does the sleep drive have such preemptive effects on behavior and performance? Formulating better answers to these simple questions is the purpose of this proposal. We use an animal model in our quest to understand the mechanisms mediating the homeostatic sleep drive (HSD), the drive that mediates the sleepiness following sleep loss or sleep deprivation (SD). Our earlier findings in animal models pointed to elevated extracellular levels of the neurochemical adenosine (AD) as a homeostatic sleep factor acting potently in the cholinergic basal forebrain (CBF) to increase sleepiness after as little as 3 to 6h of sleep deprivation (short term SD). We now think that with progressively longer periods of SD (6 or more h of SD), the HSD effects are progressively more mediated by non-CBF cortical regions. We note that "adenosine and sleep" is a scientific hot topic, with a February 2008 PubMed search revealing some 31 articles in the past year alone. To help advance our understanding, we propose the following studies relevant to the molecular, cellular, and behavioral mechanisms underlying the HSD. Specific Aim 1). We will test whether changes in AD production and extracellular levels occur first in CBF and then, with longer SD, in cortical areas. We predict an increase in adenosine A1 receptor binding (number of active receptors) with 24h SD. We will test our preliminary data-derived hypothesis indicating that inducible nitric oxide synthase (iNOS)-produced nitric oxide (NO) is an immediate mediator of AD increases during SD, and we predict that iNOS and NO activity will show the same temporal-spatial distribution as AD. Specific Aim 2). Using in vitro slice in a novel genetically modified mouse that expresses green fluorescent protein (GFP) in GABAergic neurons, we will investigate the cellular actions of NO to confirm preliminary data that NO initially excites cortically projecting CBF cholinergic and GABAergic neurons, and then produces an inhibition that is dependent on AD produced by NO. We further will study the intrinsic neurophysiological properties and pharmacology of identified cellular components of the CBF. Specific Aim 3). We will investigate the effects and interaction of SD, AD and NO in the CBF on sleepiness and vigilance using a novel rodent version of the human multiple sleep latency test (rMSLT) and a rodent version of the human psychomotor vigilance task (rPVT). In summary, this project proposes to use what we view as state-of-the-art molecular, electrophysiological and behavioral methods, including several which are unique to our lab within the sleep field. These include the use of a novel dye (DAF) to identify the cellular sources of NO production; the use of genetically modified GFP mice which allow identification of GABAergic neurons prior to electrophysiological recordings and finally the use of rodent analogues of human tests measuring sleepiness (rMSLT) and vigilance (rPVT). We thus see this application as using state-of-the art technology to answer critical questions about the homeostatic sleep drive. PUBLIC HEALTH RELEVANCE Sleep loss from disease (including insomnia and depression) and vocational demands (shift work, doctors, emergency workers) is known as a major contributor to diminished quality of life, to accidents and to decreased work and school efficiency. This proposal uses an animal model to enable understanding of the biological mechanisms mediating the sleepiness following sleep loss or sleep deprivation. By doing this we hope to lay the foundation for a rational treatment of insomnia and other sleep disorders.

描述（申请人提供）：为什么我们醒后会犯困？为什么睡眠内驱力会对行为和表现产生如此先发制人的影响？为这些简单问题制定更好的答案是本提案的目的。我们使用动物模型来了解介导稳态睡眠驱动力 (HSD) 的机制，这种驱动力介导睡眠不足或睡眠剥夺 (SD) 后的嗜睡。我们早期在动物模型中的发现表明，神经化学物质腺苷 (AD) 的细胞外水平升高，作为一种稳态睡眠因子，在胆碱能基底前脑 (CBF) 中发挥有效作用，在睡眠不足 3 至 6 小时后（短期 SD ）。我们现在认为，随着 SD 时间逐渐延长（SD 6 小时或更长），HSD 效应逐渐更多地由非 CBF 皮质区域介导。我们注意到“腺苷和睡眠”是一个科学热门话题，2008 年 2 月的 PubMed 搜索仅在过去一年就显示了大约 31 篇文章。为了帮助加深我们的理解，我们提出了以下与 HSD 的分子、细胞和行为机制相关的研究。具体目标 1).我们将测试 AD 产生和细胞外水平的变化是否首先发生在 CBF 中，然后在较长的 SD 中发生在皮质区域。我们预测 24 小时 SD 时腺苷 A1 受体结合（活性受体的数量）会增加。我们将测试我们的初步数据衍生假设，表明诱导型一氧化氮合酶 (iNOS) 产生的一氧化氮 (NO) 是 SD 期间 AD 增加的直接介质，并且我们预测 iNOS 和 NO 活性将表现出相同的时空变化分布为AD。具体目标2)。使用在 GABA 能神经元中表达绿色荧光蛋白 (GFP) 的新型转基因小鼠的体外切片，我们将研究 NO 的细胞作用，以确认初步数据，即 NO 最初会兴奋皮质投射的 CBF 胆碱能和 GABA 能神经元，然后产生依赖于 NO 产生的 AD 的抑制作用。我们将进一步研究 CBF 的已识别细胞成分的内在神经生理学特性和药理学。具体目标 3)。我们将使用新型啮齿动物版本的人类多次睡眠潜伏期测试 (rMSLT) 和啮齿动物版本的人类精神运动警觉任务 (rPVT) 来研究 CBF 中的 SD、AD 和 NO 对嗜睡和警觉性的影响和相互作用。总之，该项目建议使用我们认为最先进的分子、电生理学和行为方法，包括我们实验室在睡眠领域独有的几种方法。其中包括使用新型染料 (DAF) 来识别 NO 产生的细胞来源；使用转基因 GFP 小鼠，可以在电生理记录之前识别 GABA 能神经元，最后使用人类测试的啮齿动物类似物测量困倦 (rMSLT) 和警觉性 (rPVT)。因此，我们认为该应用程序使用最先进的技术来回答有关稳态睡眠驱动的关键问题。公共卫生相关性 疾病（包括失眠和抑郁）和职业需求（轮班工作、医生、急救人员）导致的睡眠不足被认为是导致生活质量下降、事故发生以及工作和学校效率下降的主要原因。该提案使用动物模型来了解调节睡眠不足或睡眠剥夺后嗜睡的生物机制。通过这样做，我们希望为合理治疗失眠和其他睡眠障碍奠定基础。