Circuit-specific molecular mechanisms in fentanyl use and relapse

项目总结/摘要 药物成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。电流 药物滥用治疗并非对所有人都有效，许多正在康复的吸毒者继续复发。药物 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 (VTA)和丘脑核（NAc）。NAc和VTA都经历分子和生理变化， 对滥用药物的反应。腹侧被盖区将多巴胺能投射发送到NAc，NAc将GABA能投射发送到NAc。 投影回到VTA。NAc和VTA的活性是驱动药物使用的关键， 毒瘾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，从NAc回到VTA的连接在药物成瘾中研究不足。 阿片类药物使用、阿片类药物成瘾和阿片类药物过量死亡的发生率处于前所未有的高位。在 在K99/R 00独立之路奖中，我的目标是确定NAc对VTA的预测如何影响阿片类药物 使用和复发。使用芬太尼自我给药的小鼠模型，我收集了初步数据，显示 投射到腹侧被盖区的NAc神经元控制芬太尼在强迫戒断后的寻求。在这项研究中，我打算利用 交叉电路操作和尖端分子生物学来研究分子机制 NAc投射到VTA控制芬太尼摄入和复发。在目标1的指导阶段，我将 学习操作性自我管理程序，并将其与化学遗传操作结合联合收割机，以评估 VTA中NAc终末在芬太尼使用和复发中的必要性在目标2中，我将学习RNAseq和原位 杂交以分析芬太尼自身给药后特定VTA神经元中的分子适应。期间 在独立阶段，在目标3中，我将使用新的病毒构建体来评估分子操作的作用 芬太尼使用和复发中特定腹侧被盖区神经元的变化。在目标4中，我将评估芬太尼和分子的作用， 使用快速扫描循环伏安法对NAc中多巴胺释放的操作。在一起，研究 在这个独立之路奖中提出的将阐明NAc-VTA回路中的分子机制 推动阿片类药物的使用和复发，同时为我提供必要的工具， 独立的研究计划，桥梁分子生物学，电路操纵，操作行为， 伏安法用于检测成瘾。

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.