Role of lateral habenula in methamphetamine TAAR1-mediated synaptic plasticity and aversion

外侧缰核在甲基苯丙胺 TAAR1 介导的突触可塑性和厌恶中的作用

• 批准号: 10733665
• 负责人: Susan L Ingram
• 金额: $ 56.56万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10733665
• 关键词: Ablation; Address; Agonist; Amines; Behavior; Behavioral; Bilateral; Brain; Breeding; Cell Nucleus; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Consumption; Data; Dependence; Disease; Dopamine; Dorsal; Dose; Drug Use Disorder; Electrophysiology (science); G-Protein-Coupled Receptors; Genetic; Genotype; Glutamate Receptor; Glutamate Transporter; Glutamates; Goals; Habenula; Injections; Intake; Knock-in; Knock-in Mouse; Label; Laboratories; Lateral; Lesion; Mediating; Mediation; Membrane; Membrane Transport Proteins; Methamphetamine; Methamphetamine dependence; Motivation; Mus; Neurons; Pathway interactions; Pattern; Pharmaceutical Preparations; Physiological; Presynaptic Terminals; Property; Publishing; Regulation; Research; Rewards; Risk Reduction; Role; Serotonin; Serotonin Agonists; Signal Transduction; Slice; Synapses; Synaptic plasticity; Testing; Therapeutic; Tracer; Ventral Tegmental Area; Wild Type Mouse; With laterality; behavioral study; dopamine transporter; dopaminergic neuron; effective therapy; excitatory amino acid transporter 3; experience; extracellular; in vivo; methamphetamine action; methamphetamine effect; methamphetamine use; monoamine; neural circuit; neurotransmitter release; optogenetics; patch clamp; presynaptic; receptor; retrograde transport; serotonin transporter; tool

PROJECT SUMMARY Considerable research has focused on drug use disorders as motivational disorders involving inherent or drug- induced reward pathway function. However, the focus of this application is on opposing aversive effects of methamphetamine (MA) that may curb its use, which have been little studied. The Richards (Phillips) laboratory identified the trace amine associated receptor 1 (TAAR1) as a critical target impacting MA-induced aversion. TAAR1 is an intracellularly located G protein-coupled receptor. MA gains access to TAAR1 only if it is transported into the cell. This occurs via extracellular membrane transporters, such as dopamine and serotonin transporters, DAT and SERT, respectively. We propose that TAAR1 activation by MA in dopamine and serotonin neurons is responsible for MA-induced aversion and that monoaminergic circuit interactions with the lateral habenula (LHb) are of particular importance. The overarching goal of the studies proposed in this application is to understand the monoaminergic neuron-LHb interactions responsible for the experience of MA- induced aversion via TAAR1 that may reduce risk for MA use. Our preliminary data show that MA activates lateral habenula (LHb) neurons, specifically in mice with functional TAAR1. The studies in Aim 1 will use slice electrophysiology to examine the TAAR1-dependent effects of MA in ventral tegmental area dopamine and dorsal raphe serotonin neurons, comparing slices from wildtype and CRISPRed knock-in mice, mice that have functional vs. nonfunctional TAAR1, respectively. Aim 1 studies will also use optogenetic stimulation to examine the effects of MA on glutamatergic synapses from the LHb. Based on our published findings, functional behavioral studies in this aim will examine the role of a glutamate receptor subunit, GluN2B, on MA aversion and intake. Aim 2 will perform behavioral studies focused on the LHb, which has been shown to mediate other types of aversion, has not been studied for MA aversion. We will ablate the LHb in mice with and without functional TAAR1 and study the impact on MA aversion and intake. Finally, Aim 3 studies will use a retrograde tracer to identify the LHb neurons that project to either the ventral tegmentum or dorsal raphe. Electrophysiological studies will determine whether dopamine or serotonin modulate MA activation of LHb neurons and determine whether MA activation of TAAR1 in presynaptic terminals of ventral tegmental area dopamine neurons or dorsal raphe serotonin neurons regulate the effects of MA using slices from mice with and without functional TAAR1. Thus, this proposal utilizes genetic tools, circuit analysis via electrophysiology and behavioral analysis to identify how MA engages LHb neurons in a TAAR1-dependent manner, whether LHb circuits are necessary for MA-induced aversion behaviors, and whether they inhibit MA intake. This strategy could be used to study effects of MA on TAAR1 signaling in other regions. The study of mechanisms underlying sensitivity to MA-induced aversion could lead to the identification of a new class of therapeutics.

项目概要 大量研究集中在药物使用障碍作为涉及固有或药物的动机障碍。 诱导奖赏通路功能。然而，本申请的重点是反对 甲基苯丙胺（MA）可能会限制其使用，但对此的研究很少。理查兹 (菲利普斯) 实验室确定微量胺相关受体 1 (TAAR1) 是影响 MA 诱导的关键靶点 厌恶。 TAAR1 是一种位于细胞内的 G 蛋白偶联受体。仅当 MA 获得对 TAAR1 的访问权时， 被输送到细胞内。这是通过细胞外膜转运蛋白发生的，例如多巴胺和 血清素转运蛋白分别为 DAT 和 SERT。我们认为多巴胺中的 MA 可以激活 TAAR1 血清素神经元负责 MA 诱导的厌恶以及单胺能回路与 外侧缰核 (LHb) 特别重要。本研究提出的总体目标 应用程序是为了了解单胺能神经元 - LHb 相互作用，负责 MA- 的体验 通过 TAAR1 诱导厌恶，可能会降低使用 MA 的风险。我们的初步数据显示 MA 激活 外侧缰核 (LHb) 神经元，特别是在具有功能性 TAAR1 的小鼠中。目标 1 中的研究将使用切片 电生理学检查 MA 对腹侧被盖区多巴胺和 TAAR1 依赖性的影响 中缝背侧血清素神经元，比较野生型和 CRISPR 敲入小鼠的切片， 分别是功能性 TAAR1 和非功能性 TAAR1。目标 1 研究还将利用光遗传学刺激 检查 MA 对 LHb 谷氨酸能突触的影响。根据我们发表的调查结果， 此目的的功能行为研究将检查谷氨酸受体亚基 GluN2B 对 MA 的作用 厌恶和摄入。目标 2 将进行针对 LHb 的行为研究，该研究已被证明可以 调解其他类型的厌恶，尚未针对 MA 厌恶进行研究。我们将用以下方法消融小鼠的 LHb： 以及没有功能性 TAAR1 并研究对 MA 厌恶和摄入的影响。最后，Aim 3 研究将使用 一种逆行示踪剂，用于识别投射到腹侧被盖或中缝背侧的 LHb 神经元。 电生理学研究将确定多巴胺或血清素是否调节 LHb 的 MA 激活 神经元并确定腹侧被盖区突触前末梢 TAAR1 的 MA 激活是否 多巴胺神经元或中缝背侧血清素神经元使用小鼠切片调节 MA 的作用 并且没有功能性 TAAR1。因此，该提案利用遗传工具，通过电生理学进行电路分析 和行为分析以确定 MA 如何以 TAAR1 依赖性方式参与 LHb 神经元，是否 LHb 回路对于 MA 引起的厌恶行为以及它们是否抑制 MA 的摄入是必需的。这 策略可用于研究 MA 对其他区域 TAAR1 信号传导的影响。机制研究 对 MA 引起的厌恶的潜在敏感性可能会导致一类新疗法的鉴定。