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.

项目摘要 大量的研究集中在药物使用障碍上，作为涉及固有或药物的动机疾病 诱发奖励途径功能。但是，此应用的重点是相反的厌恶作用 甲基苯丙胺（MA）可能会遏制其使用，这几乎没有研究。理查兹（菲利普斯） 实验室确定痕量胺相关受体1（TAAR1）是影响MA诱导的关键靶标 厌恶。 TAAR1是一种细胞内的G蛋白偶联受体。 MA仅在TAAR1的情况下获得访问权限 被运输到细胞中。这是通过细胞外膜转运蛋白（例如多巴胺和）发生的 5-羟色胺转运蛋白，DAT和SERT。我们建议在多巴胺中通过MA激活TAAR1 5-羟色胺神经元负责MA诱导的厌恶，并与单胺能回相互作用 外侧Habenula（LHB）特别重要。在此提出的研究的总体目标 应用是了解单胺能神经元LHB相互作用，负责MA-的经验 通过TAAR1引起的厌恶可能会降低MA使用的风险。我们的初步数据显示MA激活 外侧Habenula（LHB）神经元，特别是在具有功能性TAAR1的小鼠中。 AIM 1中的研究将使用切片 电生理，以检查MA在腹侧段区域多巴胺和 背侧raphe 5-羟色胺神经元，比较了野生型和Crispred敲入小鼠的切片，具有 功能和非功能性TAAR1。 AIM 1研究还将使用光遗传刺激 检查MA对LHB的谷氨酸能突触的影响。根据我们已发表的发现， 此目标中的功能行为研究将检查谷氨酸受体亚基Glun2b在MA上的作用 厌恶和进气。 AIM 2将进行专注于LHB的行为研究，已证明 介导其他类型的厌恶，尚未研究以进行MA厌恶。我们将在老鼠中消灭LHB 并且没有功能性TAAR1并研究对MA厌恶和摄入的影响。最后，AIM 3研究将使用 逆行示踪剂，可识别向腹侧刺或背侧raphe投射的LHB神经元。 电生理研究将确定多巴胺或5-羟色胺是否调节LHB的MA激活 神经元并确定在腹侧对段区域突触前末端中TAAR1的MA激活是否激活 多巴胺神经元或背侧raphe 5-羟色胺神经元使用小鼠的切片调节MA的作用 并且没有功能性taar1。因此，该建议利用遗传工具，通过电生理学分析 和行为分析以识别MA如何以TAAR1依赖性方式与LHB神经元相关 LHB电路对于MA引起的厌恶行为是必需的，以及它们是否抑制MA摄入量。这 策略可用于研究MA对其他区域中TAAR1信号传导的影响。机制的研究 对MA诱导的厌恶的潜在敏感性可能导致鉴定新的治疗剂。