Determining if Activity in Specific Lateral Habenula Output Pathways Motivates Avoidance of Synthetic Opioid Withdrawal or Cue Induced Reinstatement

项目摘要 阿片类药物滥用在美国已达到流行病的程度， 每年死亡人数1人。特别是，合成阿片类药物成瘾已被证明是非常困难的， 战斗，部分原因是它在用户中生成强大的对手进程。每剂合成阿片类药物 产生快速和强烈的欣快，这是强烈相关的药物线索，而退出和禁欲， 会导致严重的精神痛苦避免戒断和随后暴露于药物线索 是长期禁欲的主要障碍。外侧缰核（Lateral Habenula，LHb）是一个激动人心的研究对象 神经元促进复发，因为LHb活性与应激逃避和 激励价值6.急性戒断期间的LHb活性可能通过避免压力而激发复发 机制，而在禁欲期间，通常与药物配对的线索没有得到回报， 在LHb。这种活动可能会通过类似于奖励预测错误的过程来激励药物寻求。下 John Neumaier博士、Michael Bruchas博士、Paul菲利普斯博士和Charles Chavkin博士的主要指导，本K99/R 00 独立之路奖将使我能够获得尖端的体内钙成像培训， 光遗传学和口服自我给药模型开发。这次培训将使我能够阐明 主要的LHb输出途径在激励避免芬太尼戒断和线索恢复。期间 在该资助的指导阶段，我将接受培训，使用基于GCaMP 6的体内钙成像来记录LHb 特异性投射到腹侧被盖区（VTA）、嘴内侧被盖（RMTg）或 中缝背核（DRN）在纳洛酮催促戒断，条件性位置厌恶测试， 恢复芬太尼我还将接受联合收割机体内钙成像与红移光遗传学相结合的培训。 抑制，以利用同步电路成像和记录，并测试因果关系之间 神经通路活动和行为。最后，我将接受新型口服芬太尼自我给药（SA）模型的培训 这对我作为一名行为神经科学家的独立研究生涯将是无价的。期间 在该奖项的独立阶段，我将联合收割机与我以前的专业知识，以确定是否活动， 每个LHb通路对于与戒断相关的负面影响，戒断诱导的CPA， 或暗示恢复芬太尼寻求。我将通过同时抑制LHb投射和记录 从每个LHb靶区域（VTA、RMTg或DRN）中提取，以确定抑制对行为的影响， 单胺能核活性。总之，这项独立之路奖提出的研究 将阐明个体LHb通路在激发避免戒断和线索恢复中的作用， 以及它们在表达负面情绪中的作用。该奖项将提供新技术工具的培训， 资助写作，实验室管理，指导和其他必要的技能，以建立一个独立的研究 该计划能够产生高影响力的研究和培训下一代科学家。

Project Summary Opioid abuse has reached epidemic proportions in the United States and is responsible for more than 40,000 overdose deaths each year 1. In particular, synthetic opioid addiction has proven to be extremely difficult to combat, in part because it generates powerful opponent processes in the user. Each dose of synthetic opioid produces rapid and potent euphoria that is strongly associated to drug cues, while withdrawal and abstinence from synthetic opioids induce severe distress. Avoidance of withdrawal and subsequent exposure to drug cues are key deterrents to long-term abstinence. The Lateral Habenula (LHb) is an exciting target for studying neuronal facilitation of relapse, as LHb activity is correlated with both stress evasion and the encoding of motivational value 6. LHb activity during acute withdrawal may motivate relapse via stress avoidance mechanisms, while during abstinence cues that are normally paired with drugs go unrewarded, inducing activity in the LHb. This activity may motivate drug seeking via a process akin to reward prediction error. Under the primary mentorship of Drs. John Neumaier, Michael Bruchas, Paul Phillips, and Charles Chavkin, this K99/R00 Pathway to Independence award will allow me to obtain training in cutting edge in-vivo calcium imaging and optogenetics, and oral self-administration model development. This training will allow me to elucidate the roles of the major LHb output pathways in motivating avoidance of fentanyl withdrawal and cued reinstatement. During the mentored phase of this grant I will be trained to use GCaMP6-based in-vivo calcium imaging to record LHb neurons that project specifically to the ventral tegmental area (VTA), the rostromedial tegmentum (RMTg), or the dorsal raphe nucleus (DRN) during naloxone-precipitated withdrawal, conditioned place aversion testing, and fentanyl reinstatement. I will also be trained to combine in-vivo calcium imaging with red-shifted optogenetic inhibition in order to leverage simultaneous circuit imaging and recording, and test causal relationships between neural pathway activity and behavior. Finally, I will be trained in novel oral fentanyl self-administration (SA) model development, which will be invaluable to my independent research career as a behavioral neuroscientist. During the independent phase of the award, I will combine this training with my prior expertise to determine if activity in each LHb pathway is necessary for expression of withdrawal related negative affect, withdrawal induced CPA, or cued reinstatement to fentanyl seeking. I will do so by simultaneously inhibiting LHb projections and recording from each LHb target region (VTA, RMTg, or DRN) to determine the effect of inhibition on behavior and monoaminergic nucleus activity. In summary, the research proposed in this Pathway to Independence Award will elucidate the role of individual LHb pathways in motivating avoidance of withdrawal and cued reinstatement, as well as their role in the expression of negative affect. This award will provide training in new technical tools, grant writing, lab management, mentorship, and other skills necessary to establish an independent research program capable of producing high impact studies and training the next generation of scientists.