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程度，造成4万多人死亡 每年过量死亡1.尤其是合成阿片成瘾已被证明是极其困难的 战斗，部分是因为它在用户中生成了强大的对手进程。每一剂量合成阿片类药物 产生快速而强烈的欣快感，这种快感与药物线索密切相关，而戒断和禁欲 合成阿片类药物会导致严重的痛苦。避免戒毒和随后接触到毒品线索 是长期禁欲的关键威慑因素。外侧缰核(LHb)是一个令人兴奋的研究对象 复发的神经元易化，因为LHb活性与应激逃避和编码 动机价值6.急性戒断期间LHB的活动可能通过避免压力而促使复发 机制，而在禁欲期间，通常与药物配对的提示没有回报，诱导活动 在LHB。这一活动可能会通过类似于奖励预测错误的过程来激发寻求毒品的动机。在.之下 约翰·诺迈尔博士、迈克尔·布鲁查斯博士、保罗·菲利普斯博士和查尔斯·查夫金博士的主要指导，这是K99/R00 独立之路奖将使我获得体内尖端钙成像和 光遗传学和口腔自我给药模式的发展。这次培训将使我能够阐明这些角色 在激励避免芬太尼停药和提示恢复方面的主要LHb输出途径。在.期间 这笔赠款的指导阶段我将接受培训，使用基于GCaMP6的体内钙成像来记录LHb 专门投射到腹侧被盖区(VTA)、旋转内侧被盖区(RMTg)或 中缝背核(DRN)在纳洛酮催促戒断、条件性位置厌恶试验和 恢复芬太尼。我还将接受培训，将体内钙成像与红移光遗传相结合 抑制，以便利用同时的电路成像和记录，并测试 神经通路的活动和行为。最后，我将接受新的口服芬太尼自我给药(SA)模式的培训 这对我作为一名行为神经学家的独立研究生涯将是无价的。在.期间 在奖项的独立阶段，我将结合这次培训和我以前的专业知识来确定活动是否在 每条LHb通路都是戒断相关负性情绪、戒断诱导的CPA、 或暗示复职以寻求芬太尼。我将通过同时抑制LHb投影和录制来做到这一点 从每个LHb靶区(VTA、RMTg或DRN)确定抑制对行为和 单胺能核活动。综上所述，这项独立之路奖中提出的研究 将阐明单个LHb通路在激励避免戒断和提示恢复方面的作用， 以及它们在表达负面情绪方面的作用。该奖项将提供新技术工具方面的培训， 授予写作、实验室管理、指导和其他建立独立研究所需的技能 能够产生高影响力的研究和培训下一代科学家的计划。

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.