Unraveling the Locus Coeruleus Circuitry in Opioidinduced Sleep Disturbances

项目总结 本培训计划的主要目标是：1)为李詈提供医学博士和博士学位，他是麻省理工学院的执业麻醉师 拥有额外的科学和专业技能，以促进他过渡成为一名 独立研究人员，以及2)更好地了解蓝斑(LC)是如何在细胞和回路中 水平，导致阿片类药物引起的睡眠障碍。研究阿片类药物引起的睡眠障碍 重要是因为阿片类药物具有巨大的社会影响，2018年造成超过4.6万人过量死亡， 美国每年近800亿美元的经济成本；因为最近的研究表明 阿片类药物引起的睡眠障碍的几个方面，包括认知能力下降，疼痛耐受性下降， 以及更多的负面影响，导致长期使用阿片类药物和复发。有趣的是，LC和 脑室旁丘脑(PVT)是调节睡眠-觉醒平衡和调节戒断反应的两个大脑区域 阿片类药物的行为，但它们在阿片类药物诱导的睡眠障碍中的作用相对未知。因此，在 在他的K99期，李博士的目标是1)研究LC神经元活动与睡眠模式的关系 急性和慢性阿片类药物给药和2)确定局部阿片受体在LC中的作用。 诱发睡眠障碍。此外，他计划接受1光子显微内窥镜成像方面的培训， 阿片受体的光遗传操作和细胞特异性药物靶向使用的药物受到严重限制 Tethering(DART)；并将由世界著名的阿片类药物专家迈克尔·布鲁查斯教授指导 神经生物学，以及神经回路分析、睡眠和物质滥用障碍专家委员会。 总之，他的培训将帮助他过渡到独立的R00阶段，在那里他计划使用他训练有素的技能 1)检测PVT神经元活动与急性和慢性阿片类药物给药后睡眠模式的关系，以及 确定局部阿片受体在PVT中在阿片诱导的睡眠障碍中的作用。在美国的研究 独立阶段将为他未来的R01提交做好准备，以进一步检查唤醒神经回路 对镇静剂和麻醉药的反应。因此，该建议旨在描述LC-PVT电路的特性 用尖端工具建立小鼠急性和慢性阿片类药物给药模型 细胞特异性药理学，以更好地了解阿片类药物潜在的细胞和电路机制 诱导睡眠障碍，并为李博士提供必要的培训和指导，以帮助他实现 他的职业目标是创办自己的实验室，成为一名独立的调查员。

PROJECT SUMMARY The main goals of this training proposal is 1) to provide Li Li, MD, PhD, a practicing anesthesiologist at the University of Washington, with additional scientific and professional skills to facilitate his transition to become an independent investigator, and 2) to better understand how the locus coeruleus (LC), at the cellular and circuit levels, contributes to opioid-induced sleep disturbances. Studying sleep disturbances caused by opioids is important because opioids have enormous societal impact, causing over 46,000 overdose deaths in 2018 and almost $80 billion in economic cost annually in the United States; and because recent studies have implicated several aspects of sleep disturbances caused by opioids, including poorer cognition, decreased pain tolerance, and increased negative affect, in contributing to chronic opioid use and relapse. Interestingly, the LC and paraventricular thalamus (PVT) are two brain regions that modulate sleep-wake balance and mediate withdrawal behaviors from opioids, but their roles in opioid-induced sleep disturbances are relatively unknown. Thus, during his K99 phase, Dr. Li aims to 1) examine LC neuronal activity in relation to sleep patterns in a mouse model of acute and chronic opioid administration and 2) determine the role of local opioid receptors in the LC in opioid- induced sleep disturbances. Furthermore, he plans to receive training in 1-photon microendoscopic imaging, optogenetic manipulation of opioid receptors, and cell-specific drug targeting using drug acutely restricted by tethering (DART); and will be mentored by Prof. Michael Bruchas, a world-renowned expert in opioid neurobiology, and a committee of experts in neural circuitry analysis, sleep, and substance abuse disorders. Together, his training will help transition to his independent R00 phase, where he plans to use his trained skills 1) to examine PVT neuronal activity in relation to sleep patterns in acute and chronic opioid administration, and to determine role of local opioid receptors in PVT in opioid-induced sleep disturbances. Studies in the independent phase will prepare him for a future R01 submission to further examine arousal neural circuitry response to sedative and anesthetic drugs. Thus, this proposal aims to characterize the LC-PVT circuit in a mouse model of acute and chronic opioid administration using cutting-edge tools in neural circuit analysis and cell-specific pharmacology in order to better understand the cellular and circuit mechanism underlying opioid- induced sleep disturbances, and provides the essential training and mentorship for Dr. Li to help him achieve his career goal of starting his own lab and becoming an independent investigator.