SYNAPTIC BASIS OF SLEEP CYCLE CONTROL

睡眠周期控制的突触基础

• 批准号: 6391883
• 负责人: Robert W McCarley
• 金额: $ 39.93万
• 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 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6391883
• 关键词: REM sleep; acetylcholine; adenosine; biocytin; brain electrical activity; cats; dorsal raphe nucleus; electroencephalography; electromyography; electrooculography; hypothalamus; laboratory rat; microdialysis; neuropharmacology; preoptic areas; prosencephalon; serotonin receptor; sleep; sleep regulatory center; stereotaxic techniques; wakefulness

The broad purpose of this series of studies is to use work in animals to understand the physiological and pharmacological mechanism controlling sleep, and thereby provide a sound baiss for the understanding and treatment of human sleep disorders, both primary and secondard to medical and physchiatric conditions. Previous work has greatly advanced our knowledge of brainstem mechanisms controlling the rapid eye movement (REM) phase of sleep, suggesting that brainstem neurons using acetylcholine as a neurotransmitter (cholinergic neurons) promote this phase of sleep. This application builds on, and extends this work. The key techniques to be used are a novel combination of microdialysis and extramcellular unit recording in freely moving cats, intracellular recordings in naturally sleeping cats and the rat in vitro slice preparation. Hypotheses to be investigated include whether increases in adenosine following prolonged wakefulness (and increased etabolic activity) act as a factor reducing wakefulness (W) and increasing the Slow Wave Sleep (SWS or nonREM) phase of sleep. We hypothesize adenosine increases act most strongly on cholinergic neurons in the basal forebrain and the mesopontine area that promote W and an activated electroencephalogram (EEG). New data suggest strong adenosine state-altering effects on the dorsal raphe nucleus (DRN) also. Microdialysis measurements of extracellular adenosine and delivery of adenosine transport inhibitors and concomitant unit recordings will be used for in vivo tests, while in vitro studies will examine mechanisms of action. The hypothesis that serotonin-containing DRN neurons disinhibit cholinergic neurons and allow REM sleep to occur when these DRN neurons slow discharge during SWS and REM will also be evaluated; in vivo and in vitro techniques will examine the degree to which each of four factors may control the slowing of DRN discharge: GABA, 5HT collateral feedback, adenosine, and presynaptic disfacilitation of adrenergic input. We will use intracellular in vivo recording and double labeling to determine if the REM-on neurons (=discharge activity selective for REM sleep, and possibly controlling this state) and the Waking- and REM-on neurons (W/R-on, possibly controlling EEG activation in both W and REM) recorded in the mesopontine cholinergic zone can be positively identified as cholinergic. Using microdialysis and unit recording we will test the hypothesis that the REM-on neurons of this cholinergic zone differ from W/R-on neurons by their inhibtability by DRN input acting on 5HT1A receptors. Finally we will examine the Ventrolateral preoptic Area (VLPOA) of hypothalamus, where earlier work with cFos protein indicated a selective activation of a population of neurons during SWS. Using microdialysis (MD) we will evaluate whether spontaneous extracellular GABA levels decrease during SWS, suggesting disinhibition, and whether MD-perfused bicuculline promotes SWS, as it did in preliminary data. We will also evaluate the extent of cholinergic control (from basal forebrain) and of histaminergic control (from the tuberomammillary nucleus). In vitro work will examine the post- and pre-synaptic effects of these and other neurotransmitters on VLPOA neurons identified with biocytin and GAD immunohistochemical labeling.

这一系列研究的广泛目的是将动物的作品用于 了解生理和药理机制 控制睡眠，从而为 理解和治疗人类睡眠障碍，两者都是主要的 第二次是医学和医学疾病。 以前的工作 大大提高了我们对脑干机制的了解 控制睡眠的快速眼动（REM）阶段，表明 使用乙酰胆碱作为神经递质的脑干神经元 （胆碱能神经元）促进这一睡眠阶段。 此应用程序 建立并扩展这项工作。 要使用的关键技术是 微透析和细胞外单元记录的新型组合 在自由移动的猫中，自然睡猫中的细胞内记录 和大鼠体外切片制剂。 要研究的假设包括腺苷的增加是否增加 在长时间的清醒（以及依代代谢活性增加）ACT之后 作为降低清醒（W）并增加慢波的因素 睡眠（SWS或非REM）睡眠阶段。 我们假设腺苷 增加对基底胆碱能神经元的作用最强烈 前脑和促进W和激活的中桥区域 脑电图（EEG）。 新数据表明腺苷强 改变状态对背侧raphe核（DRN）的影响。 细胞外腺苷的微透析测量和递送 腺苷转运抑制剂和随之而来的单位记录将是 用于体内测试，而体外研究将检查机制 行动。 含5-羟色胺DRN神经元的假设抑制了 胆碱能神经元，并在这些DRN时允许REM睡眠 神经元在SWS和REM期间的缓慢排出也将评估； 体内和体外技术将检查每个程度 在四个因素中，可能控制DRN放电的减慢：GABA， 5HT附带反馈，腺苷和突触前的异病毒 肾上腺素能输入。 我们将使用细胞内的体内记录和 双重标记以确定是否恢复神经元（=放电） 选择性睡眠选择性，可能控制此状态） 以及醒来的神经元和恢复神经元（w/r-ON，可能控制 在中桥中记录的W和REM中的脑电图激活 胆碱能带可以正面识别为胆碱能。 使用 微透析和单位记录我们将检验以下假设。 该胆碱能区的恢复神经元与W/R-ON神经元不同 通过其可约束性通过作用于5HT1A受体的DRN输入。 最后，我们将检查腹外侧前区域（VLPOA） 下丘脑，早期使用CFOS蛋白表明 SWS期间神经元种群的选择性激活。 使用 微透析（MD）我们将评估自发细胞外 SWS期间的GABA水平降低，表明抑制作用，并 像MD完美的双瓜氨酸是否促进了SWS 初步数据。 我们还将评估胆碱能的程度 对照（来自基础前脑）和组胺能控制（来自 结核核）。 体外工作将检查后和 这些和其他神经递质对VLPOA的突触前作用 用生物细胞和GAD免疫组织化学鉴定的神经元 标签。

项目成果

Sleep and brain energy levels: ATP changes during sleep.

• DOI: 10.1523/jneurosci.1423-10.2010
• 发表时间: 2010-06-30
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Dworak M;McCarley RW;Kim T;Kalinchuk AV;Basheer R
• 通讯作者: Basheer R

The role of cholinergic basal forebrain neurons in adenosine-mediated homeostatic control of sleep: lessons from 192 IgG-saporin lesions.

• DOI: 10.1016/j.neuroscience.2008.08.040
• 发表时间: 2008-11-11
• 期刊: Neuroscience
• 影响因子: 3.3
• 作者: Kalinchuk AV;McCarley RW;Stenberg D;Porkka-Heiskanen T;Basheer R
• 通讯作者: Basheer R

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• DOI: 10.5665/sleep.1110
• 发表时间: 2011
• 期刊: Sleep
• 影响因子: 5.6
• 作者: Dworak,Markus;McCarley,RobertW;Kim,Tae;Kalinchuk,AnnaV;Basheer,Radhika
• 通讯作者: Basheer,Radhika

Sleep, brain energy levels, and food intake: Relationship between hypothalamic ATP concentrations, food intake, and body weight during sleep-wake and sleep deprivation in rats.

• DOI: 10.1007/s11818-011-0524-y
• 发表时间: 2011-06
• 期刊: Somnologie : Schlafforschung und Schlafmedizin = Somnology : sleep research and sleep medicine
• 作者: Dworak M;Kim T;McCarley RW;Basheer R
• 通讯作者: Basheer R