Circuit-specific molecular mechanisms in fentanyl use and relapse

项目摘要/摘要 毒瘾是一种复杂的大脑疾病，对个人和经济健康造成巨大损失。当前 药物滥用治疗并不是对所有人都有效，许多正在康复的吸毒者继续复发。药效 暴露篡夺了正常的奖赏回路功能，包括腹侧被盖区之间的联系 伏核(VTA)和伏核(NAC)。NAC和VTA都经历了分子和生理变化 对滥用药物的反应。VTA向NAC发送多巴胺能投射，NAC向NAc发送GABA能 投射回VTA。NAC和VTA的活动是推动药物使用的关键，并与 毒品复发。许多研究已经确定了多巴胺能VTA对NAC投射的重要性， 然而，在药物成瘾中，NAC与VTA之间的联系还未得到充分研究。 阿片类药物的使用、阿片成瘾和阿片类药物过量死亡的发生率处于前所未有的高水平。在……里面 这个K99/R00独立之路奖，我的目标是确定NAC对VTA的投射如何影响阿片类药物 使用和旧病复发。使用芬太尼自我给药的小鼠模型，我收集了初步数据，显示 向VTA投射的NAC神经元控制强制戒断后的芬太尼寻求。在这项研究中，我打算利用 交叉回路操纵和前沿分子生物学研究分子机制 通过NAC预测VTA控制芬太尼的摄取和复发。在指导阶段，在目标1中，我将 学习可操作的自我给药程序，并将其与化学发生操作相结合来评估 芬太尼应用和复发时VTA中NAC终末的必要性。在目标2中，我将学习RNAseq和原位 应用杂交技术分析芬太尼自身给药后特定VTA神经元的分子适应性。在.期间 在目标3的独立阶段，我将使用新的病毒结构来评估分子操作的作用。 在特定的VTA神经元中，芬太尼的使用和复发。在目标4中，我将评估芬太尼和分子的作用 应用快速扫描循环伏安法对NAc中多巴胺释放的调控。总之，这项研究 在独立之路奖中提出的将阐明NAC-VTA电路中的分子机制 推动阿片类药物的使用和复发，同时为我提供建立 独立的研究计划，连接分子生物学、电路操纵、操作行为和 伏安法用于检测毒瘾。

Project Summary/Abstract Drug addiction is a complex brain disorder that takes an enormous toll on individual and economic health. Current drug-abuse treatments are not effective in all individuals, and many recovering addicts continue to relapse. Drug exposure usurps normal reward circuit function, including the connections between the ventral tegmental area (VTA) and the nucleus accumbens (NAc). Both NAc and VTA undergo molecular and physiological changes in response to drugs of abuse. The VTA sends dopaminergic projections to the NAc, and the NAc sends GABAergic projections back to the VTA. Activity in NAc and VTA are key for driving drug use and are heavily implicated in drug relapse. Numerous studies have established the importance of the dopaminergic VTA to NAc projection, however the connection from NAc back to VTA is understudied in drug addiction. The incidence of opioid use, opioid addiction, and death from opioid overdose are at an unprecedented high. In this K99/R00 Pathway to Independence Award, I aim to identify how NAc projections to the VTA influence opioid use and relapse. Using a mouse model of fentanyl self-administration, I have collected preliminary data showing NAc neurons that project to VTA control fentanyl seeking after forced abstinence. In this study, I intend to utilize intersectional circuit manipulation and cutting-edge molecular biology to investigate the molecular mechanisms by which NAc projections to VTA control fentanyl intake and relapse. During the mentored phase, in Aim 1, I will learn operant self-administration procedures and combine them with chemogenetic manipulation to assess the necessity of NAc terminals in VTA in fentanyl use and relapse. In Aim 2, I will learn RNAseq and in situ hybridization to profile molecular adaptations in specific VTA neurons after fentanyl self-administration. During the independent phase, in Aim 3, I will use novel viral constructs to assess the role of molecular manipulations in specific VTA neurons in fentanyl use and relapse. In Aim 4, I will assess the role of fentanyl and molecular manipulations on dopamine release in the NAc using fast-scan cyclic voltammetry. Together, the research proposed in this Pathway to Independence Award will elucidate molecular mechanisms in the NAc-VTA circuit driving opioid use and relapse, while simultaneously providing me with the tools necessary for establishing an independent research program that bridges molecular biology, circuit manipulation, operant behavior, and voltammetry for examining addiction.