Investigating the ventral pallidum-ventral tegmental area circuit in cocaine relapse

研究可卡因复发中的腹侧苍白球-腹侧被盖区回路

• 批准号: 10592313
• 负责人: Rianne Campbell
• 金额: $ 6.95万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-08 至 2025-03-07
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10592313
• 关键词: Abstinence; Affect; Area; Behavior; Behavioral Model; Biological Assay; Biosensor; Calcium; Cannulas; Career Choice; Cells; Cocaine; Cocaine misuse; Cocaine use disorder; Cue-induced relapse; Cues; Data; Development; Female; Fiber; Fiber Optics; Gene Expression; Genes; Genetic; Genetic Transcription; Globus Pallidus; Immunoprecipitation; Impairment; Implant; Individual; Intravenous; Left; Link; Measures; Mediating; Messenger RNA; Methods; Modeling; Molecular; Mus; Neurons; Pathology; Pathway interactions; Pattern; Pharmaceutical Preparations; Photometry; Population; Process; Rattus; Relapse; Research; Research Personnel; Rewards; RiboTag; Ribosomes; Saline; Science; Self Administration; Stimulus; Synapses; Synaptic Transmission; Synaptic plasticity; Techniques; Testing; Therapeutic; Therapeutic Intervention; Tissues; Training; Ventral Tegmental Area; Virus; career; cocaine craving; cocaine cue; cocaine overdose; cocaine relapse; cocaine seeking; cocaine use; design; effective therapy; experimental study; in vivo; male; neural; neuromechanism; new therapeutic target; novel; optical fiber; overdose death; prevent; response; skill acquisition; skills; transcriptome sequencing

Project Summary/Abstract: Rises in cocaine misuse and cocaine overdose deaths over the last decade illustrate the lack of therapeutic interventions for individuals with cocaine use disorder (CUD). With abstinence as the only treatment for cocaine cravings, a majority of individuals with CUD are vulnerable to relapse. The development of novel CUD therapeutics relies on a thorough understanding of the neural mechanisms that drive cocaine craving and relapse. Cocaine causes molecular adaptations within distinct neuronal subpopulations that leads to circuit- specific changes in neural activity. A critical node of the reward circuit is the ventral pallidum (VP), as it receives input from and projects to several regions that mediate cocaine-seeking behaviors. Activity from VP afferents to the ventral tegmental area (VTA-projecting VP neurons) is required for cocaine relapse; however, the cellular and molecular adaptations that occur within VTA-projecting VP neurons to promote cocaine-seeking, are unknown. To better understand the pathology of CUD, this proposal will investigate how changes in neuronal activity and gene expression within VTA-projecting VP neurons occur within cue-induced reinstatement to cocaine-seeking. Using cocaine intravenous self-administration (IVSA) and chemogenetics in mice, my preliminary data replicates a previous finding in rats that VTA-projecting VP neurons are required for reinstatement of cocaine-seeking. In this proposal, I will employ IVSA, cutting-edge in vivo biosensors, chemogenetics and molecular techniques to investigate the circuit-specific changes in VP neuronal activity and gene expression that occur during reinstatement to cocaine-seeking. I hypothesize that cocaine engages VTA- projecting VP neurons, leading to cocaine cue-induced 1) enhancement of VTA-projecting VP neuronal activity and 2) increases in synaptic gene expression that accompany drug-seeking and relapse. In Specific Aim 1, I will use fiber photometry to assess how cocaine-associated cues affected patterned calcium activity within VTA- projecting VP neurons during cue-induced reinstatement. I will also examine how chemogenetic inhibition of VTA-projecting VP neurons impacts changes in VTA-projecting VP calcium activity following the presentation of cocaine-associated cues. In Specific Aim 2, I will perform ribosomal immunoprecipitation with RNA-Sequencing to determine whether increases in synaptic gene expression within VTA-projecting VP neurons occurs following reinstatement to cocaine-seeking. Further, I will examine whether inhibiting VTA-projecting VP neuronal activity prevents reinstatement-induced changes in gene expression using chemogenetics and RNAScope. Together, these experiments will identify novel mechanisms within VTA-projecting VP neurons neurons that mediate reinstatement to cocaine-seeking. This proposal will also provide me with strong training in new research areas that will support my independent career path in science.

项目摘要/摘要： 过去十年中可卡因滥用和可卡因过量死亡人数的上升说明了治疗方法的缺乏 针对可卡因使用障碍 (CUD) 患者的干预措施。戒除可卡因是唯一的治疗方法 由于渴望，大多数患有 CUD 的人很容易复发。新型CUD的开发 治疗依赖于对驱动可卡因渴望的神经机制的透彻理解 复发。可卡因会引起不同神经元亚群内的分子适应，从而导致电路- 神经活动的具体变化。奖励回路的一个关键节点是腹侧苍白球（VP），因为它接收 来自多个调节可卡因寻求行为的区域的投入和项目。从 VP 传入到 可卡因复发需要腹侧被盖区（VTA 投射的 VP 神经元）；然而，蜂窝 VTA 投射的 VP 神经元内发生的促进可卡因寻求的分子适应是 未知。为了更好地了解 CUD 的病理学，本提案将研究如何改变 VTA 投射 VP 神经元内的神经元活动和基因表达发生在提示诱导内 恢复寻求可卡因。使用可卡因静脉自我给药（IVSA）和化学遗传学 在小鼠中，我的初步数据重复了之前在大鼠中的发现，即 VTA 投射的 VP 神经元是 恢复寻求可卡因。在这个提案中，我将采用 IVSA，尖端的体内生物传感器， 化学遗传学和分子技术研究 VP 神经元活动的回路特异性变化 恢复可卡因寻求过程中发生的基因表达。我假设可卡因参与 VTA- 投射 VP 神经元，导致可卡因诱导 1) 增强 VTA 投射 VP 神经元活动 2）伴随寻药和复发的突触基因表达增加。在具体目标 1 中，我将 使用纤维光度测定法评估可卡因相关线索如何影响 VTA 内的模式钙活动 在提示诱导恢复期间投射 VP 神经元。我还将研究化学遗传学抑制如何 VTA 投射 VP 神经元影响 VTA 投射 VP 钙活性的变化 可卡因相关的线索。在具体目标 2 中，我将使用 RNA 测序进行核糖体免疫沉淀 确定 VTA 投射 VP 神经元内突触基因表达的增加是否发生在以下情况 恢复寻求可卡因。此外，我将检查是否抑制 VTA 投射的 VP 神经元活动 使用化学遗传学和 RNAScope 防止恢复引起的基因表达变化。一起， 这些实验将确定 VTA 投射 VP 神经元内介导的新机制 恢复寻求可卡因。该提案还将为我在新的研究领域提供强有力的培训 这将支持我在科学领域的独立职业道路。