Hypothalamic Neuronal Activity During Insomnia Induced by Chronic Ethanol Exposure

在停止长期饮酒的过程中会发生严重失眠。从…获得解脱 失眠，酗酒者又开始喝酒了。虽然失眠的原因不明，但间接的 对神经元活动(c-fos，fMRI)的测量表明，在失眠期间，更多的觉醒- 促进神经元是过度活跃的。促进睡眠的神经元的状态尚不清楚，但很可能是 不那么活跃。我们的总体假设是唤醒和促进睡眠之间的活动不平衡。 神经元是戒酒期间失眠的基础。我们将直接检验这一假设。 使用新开发的神经科学工具，在经过验证的慢性乙醇中毒小鼠模型中。目标1将 使用微型示波器对特定的下丘脑觉醒促进作用中的钙离子内流(作为细胞活动的读数)进行成像 慢性间歇发作前、中、后促睡眠(GABA)神经元 酒精暴露(CIE)。我们将在清醒、非快速眼动和快速眼动睡眠中对神经元进行成像。我们假设 在失眠的CIE期，食欲素神经元更为活跃，而促进睡眠的GABA 神经元不那么活跃。我们有强有力的初步数据表明，GABA神经元已经变少 在长期饮酒后表现活跃。同样，我们的初步数据显示，食欲素神经元已经成为 更活跃。脑深部钙成像方法提供了描述性数据。因此目标2和目标3将使用 药物遗传学机械测试CIE已经改变了兴奋性阈值的假设 GABA和食欲素神经元。体外切片电生理学研究将证实这一假设。诱导睡眠 为了纠正失眠，Aim 2将激活GABA神经元，而Aim 3将抑制增食欲素神经元。 目标4将验证CIE后的假设，因为我们的药物遗传改善了睡眠 操作，那么小鼠就不会表现出更少的饮酒偏好。这一目标直指我们 改善睡眠可防止酒精复发的假说。 我们研究的总体影响是，第一次，确凿的证据将机械地将失眠联系起来 慢性酒精中毒戒断过程中特定表型活性的变化 下丘脑睡眠或促进觉醒的神经元。此外，我们的目标是翻译的，并将有助于 开发适当的失眠治疗方法。我们的目标是集成并使用尖端工具来 了解长期接触酒精是如何破坏睡眠稳态的。该项目将由以下机构联合开展 睡眠神经生物学和酒精成瘾领域的专家。

Severe insomnia occurs during the withdrawal from chronic use of alcohol. To gain relief from insomnia, alcoholics begin drinking again. Although the cause of the insomnia is unknown, indirect measurements of neuronal activity (c-FOS, fMRI) suggest that during insomnia, more of the arousal- promoting neurons are hyperactive. The status of the sleep-promoting neurons is unknown but are likely to be less active. Our overall hypothesis is that it is the imbalance in activity between wake versus sleep-promoting neurons that is the basis of insomnia during the withdrawal from alcohol. We will directly test this hypothesis in a proven mouse model of chronic ethanol intoxication using newly developed neuroscience tools. Aim 1 will use a miniscope to image Ca2+ influx (as a readout of cell activity) in specific hypothalamic wake-promoting (orexin) or hypothalamic sleep-promoting (GABA) neurons before, during and after chronic intermittent exposure to alcohol (CIE). We will image the neurons during waking, NREM, and REM sleep. We hypothesize that during the CIE-period of insomnia, the orexin neurons are more active while the sleep-promoting GABA neurons are less active. We have robust preliminary data indicating that the GABA neurons have become less active after chronic alcohol exposure. Similarly, our preliminary data reveals that orexin neurons have become more active. The deep-brain Ca2+ imaging method provides descriptive data. Thus Aims 2 and 3 will use pharmacogenetics to mechanistically test the hypothesis that CIE has shifted the excitability thresholds of the GABA and orexin neurons. In-vitro slice electrophysiology studies will confirm the hypothesis. To induce sleep to correct the insomnia, Aim 2 will activate the GABA neurons, while Aim 3 will inhibit the orexin neurons. Aim 4 will test the hypothesis that after CIE, because sleep has improved with our pharmacogenetic manipulations, then the mice will show less preference for drinking alcohol. This aim goes to the core of our hypothesis that improving sleep prevents alcohol relapse. The overall impact of our studies is that, for the first time, hard evidence will mechanistically link insomnia during the withdrawal from chronic ethanol intoxication to changes of activity in specific phenotypes of hypothalamic sleep or wake-promoting neurons. Furthermore, our aims are translational and will aid in the development of appropriate treatments for insomnia. Our aims are integrated and employ cutting-edge tools to understand how chronic ethanol exposure disrupts sleep homeostasis. This project will be conducted jointly by experts in the areas of sleep neurobiology and alcohol addiction.