Nitrergic interneurons and cue-induced cocaine seeking

氮能中间神经元和线索诱导的可卡因寻找

基本信息

批准号：
10457010
负责人：
Michael David Scofield
金额：
$ 33.97万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10457010
关键词：
Anesthesia procedures Animals Astrocytes Behavior Brain Chemosensitization Clinical Cocaine Cocaine Dependence Corpus striatum structure Cues Data Dendritic Spines Development Dopamine Dopamine D1 Receptor Dopamine D2 Receptor Dopamine Receptor Down-Regulation Drug Addiction Electrophysiology (science)Exposure to Extinction (Psychology)Genetic Glutamate Receptor Glutamate Transporter Glutamates Head Interneurons Investigation Label Link Measures Mediating Microscopic Morphology Mus Neurobiology Neurons Neurotransmitters Nitrergic Neurons Nitric Oxide Nitric Oxide Synthase Type I Nucleus Accumbens Pharmaceutical Preparations Pharmacotherapy Play Production Rattus Relapse Research Rodent Rodent Model Role Saline Signal Transduction Slice Source Sucrose Synapses Synaptic plasticity System Testing Therapeutic Therapeutic Intervention Time Training Transgenic Mice Transgenic Organisms Translating Ventral Tegmental Area Viral Viral Vector addiction cell type cocaine self-administration cohort cue reactivity design drug abstinence drug craving experimental study insight knock-down metabotropic glutamate receptor 5 neurobiological mechanism neuron loss novel novel therapeutics patch clamp prevent promoter receptor response small hairpin RNA tool vector wireless

项目摘要

PROJECT SUMMARY Vulnerability to relapse remains a significant clinical hurdle in the treatment of addiction. Data from operant rodent models of addiction and relapse indicate that cocaine-conditioned cue exposure evokes a pronounced glutamate release in the nucleus accumbens core (NAcore). Cued glutamate release engages a cell type-specific transient synaptic potentiation in NAcore medium spiny neurons (MSNs), which does not occur during cued sucrose seeking. This transient synaptic potentiation consists of increased synaptic and structural plasticity in MSNs. Importantly, the magnitude of this plasticity positively correlates with relapse behavior. We posit that both the structural component (dendritic spine head expansion) and the synaptic component (increased insertion of glutamate receptors) of this plasticity are engaged by nitric oxide (NO); a gaseous transmitter produced by interneurons that express neuronal nitric oxide synthase (nNOS). Consistent with a role for these neurons in cued relapse, we have recently demonstrated that elevated NO release in the NAcore also occurs in parallel with glutamate release during cued cocaine seeking. The objectives of the proposed study are to reveal the inputs to the NAcore required for NO release during relapse, to determine which receptors on NAcore nNOS neurons regulate NO release and cued cocaine seeking, and to reveal how nNOS participates in the induction of the transient structural and synaptic plasticity in MSNs that drives cued cocaine seeking. In Aim 1, we will use chemogenetic inhibition of inputs to the NAcore while recording glutamate and NO release during cued cocaine seeking. We predict that inputs to the NAcore carrying discreet aspects of cue and contextual salience will differentially regulate NO release and cued relapse. In Aim 2, we will elucidate how glutamate and dopamine receptors, specifically expressed on NAcore nNOS interneurons, act in concert to regulate NO production and if they are required for cued cocaine seeking. To do this we will use transgenic mice that express Cre in nNOS neurons, Cre-dependent shRNA viral vectors, as well as cocaine self-administration and cued cocaine seeking trials. We predict that glutamate and dopamine receptor systems in nNOS neurons cooperatively regulate NO release and cued cocaine seeking. In Aim 3, we will determine if loss of nNOS in the NAcore will prevent cue- mediated synaptic and structural plasticity in MSNs. To do this we will use an shRNA vector to knockdown nNOS and concomitant viral labeling of D1 or D2 receptor expressing MSNs in the NAcore. This will be done using separate cohorts of D1-and D2-promoter driven Cre rats. In Aim 3, we will measure morphological and electrophysiological readouts synaptic plasticity induced by cues. We expect that loss of nNOS will prevent both forms of plasticity linked to relapse, predominantly in D1 MSNs. In conclusion, findings from these investigations will reveal the mechanistic aspects of how nNOS neurons translate cocaine cue exposure to relapse behavior. Accordingly, preventing this relapse signal transduction by preventing cued activation of nNOS neurons represents a potential therapeutic strategy to reduce drug craving and prevent relapse.

项目摘要复发的脆弱性仍然是成瘾治疗的重大临床障碍。来自操作员的数据成瘾和复发的啮齿动物模型表明可卡因条件的提示暴露唤起了一个明显的谷氨酸在伏隔核（Nacore）中释放。提示谷氨酸释放与特定于细胞类型的特定于 Nacore培养基神经元（MSN）中的短暂性突触增强，在提示期间不会发生寻求蔗糖。这种短暂的突触增强包括增加的突触和结构可塑性 MSN。重要的是，这种可塑性的大小与复发行为呈正相关。我们认为这两个结构成分（树突状脊柱膨胀）和突触分量（增加的插入一种可塑性的谷氨酸受体由一氧化氮（NO）参与；由表达神经元一氧化氮合酶（NNOS）的中间神经元。与这些神经元中的角色一致提示复发，我们最近证明，纳科尔的升高也与谷氨酸在提示可卡因寻求期间释放。拟议研究的目标是揭示输入到在复发过程中无释放所需的nacore，以确定NACORE NNOS神经元上的哪些受体规范不释放和提示可卡因寻求，并揭示NNO如何参与归纳 MSN中的瞬态结构和突触可塑性驱动了可卡因寻求的。在AIM 1中，我们将使用在记录谷氨酸的同时，对谷氨酸的输入的化学发生抑制，在提示可卡因期间无释放寻求。我们预测，提示和上下文显着性的Nacore的投入将差异调节无释放和提示复发。在AIM 2中，我们将阐明谷氨酸和多巴胺如何专门在NACORE NNOS中间神经元中表达的受体协同起作用，以调节不产生的生产，以及if if nros中间神经元。它们是提示可卡因寻求所必需的。为此，我们将使用在NNOS中表达Cre的转基因小鼠神经元，依赖CRE的shRNA病毒载体，以及可卡因自我给药并提示可卡因寻求试验。我们预测NNOS神经元中的谷氨酸和多巴胺受体系统会调节NO 释放并提示可卡因寻求。在AIM 3中，我们将确定NACORE中NNOS的丢失是否会阻止提示 - MSN中介导的突触和结构可塑性。为此，我们将使用shRNA矢量敲除nnos 以及表达Nacore中MSN的D1或D2受体的病毒标记。这将使用单独的D1和D2启动器驱动的Cre Cre大鼠。在AIM 3中，我们将衡量形态学和通过提示引起的电生理读数突触可塑性。我们预计nnos的损失将阻止两者可塑性形式与复发有关，主要在D1 MSN中。总之，这些调查的发现将揭示NNOS神经元如何将可卡因提示暴露为复发行为的机械方面。因此，通过防止提示NNOS神经元的激活来防止这种复发信号转导代表了减少药物渴望并防止复发的潜在治疗策略。