Unraveling the Locus Coeruleus Circuitry in Opioidinduced Sleep Disturbances

项目摘要 本培训计划的主要目标是：1）为李莉，医学博士，博士，一名执业麻醉师， 华盛顿大学，具有额外的科学和专业技能，以促进他的过渡，成为一个 独立调查员，2）更好地了解蓝斑（LC），在细胞和电路 水平，有助于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 重要的是，阿片类药物具有巨大的社会影响，在2018年造成超过46，000例过量死亡， 美国每年的经济成本近800亿美元;由于最近的研究表明， 阿片类药物引起的睡眠障碍的几个方面，包括认知能力较差，疼痛耐受力下降， 和增加的负面影响，在促进慢性阿片类药物使用和复发。有趣的是，LC和 室旁丘脑（paraventricularthalamus，PVT）是调节睡眠-觉醒平衡和介导戒断的两个脑区 阿片类药物的行为，但其在阿片类药物引起的睡眠障碍的作用是相对未知的。因此，在 他的K99期，李博士的目的是1）检查LC神经元活动与睡眠模式的小鼠模型， 急性和慢性阿片样物质给药和2）确定局部阿片样物质受体在阿片样物质中的作用， 导致睡眠障碍。此外，他计划接受单光子显微内窥镜成像的培训， 阿片受体的光遗传学操纵，以及使用受 系留（DART）;并将由世界知名的阿片类药物专家Michael Bruchas教授指导 神经生物学，以及神经电路分析，睡眠和药物滥用障碍专家委员会。 总之，他的训练将有助于过渡到他的独立R 00阶段，在那里他计划使用他的训练技能 1)检查急性和慢性阿片类药物给药中与睡眠模式相关的PVT神经元活动， 以确定阿片类药物诱导的睡眠障碍中PVT中局部阿片受体的作用。的研究 独立阶段将为他将来提交R 01进一步检查唤醒神经回路做准备 对镇静剂和麻醉剂的反应。因此，该建议旨在表征LC-PVT电路中的 使用神经回路分析中的尖端工具进行急性和慢性阿片类药物给药的小鼠模型， 细胞特异性药理学，以便更好地了解阿片类药物的细胞和电路机制， 诱导睡眠障碍，并为李博士提供必要的培训和指导，以帮助他实现他的梦想。 他的职业目标是建立自己的实验室，并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.