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

• 批准号: 10630138
• 负责人: Michael David Scofield
• 金额: $ 33.98万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 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/10630138
• 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; 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; Rat Transgene; Rattus; Relapse; Research; Rodent; Rodent Model; Role; Saline; Signal Transduction; Slice; Source; Sucrose; Synapses; Synaptic plasticity; System; Testing; Therapeutic; Therapeutic Intervention; Time; Traction; Training; Transgenic Mice; Translating; Ventral Tegmental Area; Viral; Viral Vector; addiction; cell type; cocaine cue; cocaine seeking; 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; relapse prevention; response; small hairpin RNA; tool; vector; wireless

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中的一个重要临床障碍。操作数据 成瘾和复发的啮齿动物模型表明，可卡因条件线索暴露引起了明显的 谷氨酸释放的核延髓核心（NAcore）。提示谷氨酸释放参与细胞类型特异性 在NAcore中型多刺神经元（MSN）的短暂突触增强，这在提示过程中不会发生。 蔗糖搜索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， 结构成分（树突棘头部扩张）和突触成分（增加的插入） 一氧化氮（NO）是一种由谷氨酸受体产生的气体递质， 表达神经元型一氧化氮合酶（nNOS）的中间神经元。这与这些神经元在 提示复发，我们最近已经证明，在NAcore NO释放升高也发生在平行于 谷氨酸释放过程中线索可卡因寻求。拟议研究的目标是揭示投入， 复发期间NO释放所需的NAcore，以确定NAcore nNOS神经元上的哪些受体 调节NO的释放和提示可卡因寻求，并揭示nNOS如何参与诱导 MSN中的短暂结构和突触可塑性，驱动可卡因寻求线索。在目标1中，我们将使用 在提示可卡因期间记录谷氨酸和NO释放时对NAcore输入的化学发生抑制 寻找我们预测，输入到NAcore携带谨慎方面的线索和上下文的显着性将 差异调节NO释放和提示复发。在目标2中，我们将阐明谷氨酸和多巴胺 受体，特别是表达在NAcore nNOS中间神经元，共同作用，以调节NO的产生，如果 它们是寻找可卡因线索所必需的。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和线索可卡因寻求 审判我们预测nNOS神经元中的谷氨酸和多巴胺受体系统协同调节NO 释放和暗示可卡因寻求。在目标3中，我们将确定NAcore中nNOS的丢失是否会阻止提示- 介导的突触和结构可塑性。为此，我们将使用shRNA载体敲低nNOS 以及伴随的在NAcore中表达D1或D2受体的MSN的病毒标记。这将使用 D1和D2启动子驱动的Cre大鼠的单独队列。在目标3中，我们将测量形态和 电生理学读出由线索诱导的突触可塑性。我们预计，nNOS的丧失将阻止这两种情况的发生。 与复发有关的可塑性形式，主要在D1 MSN中。总之，这些调查的结果 将揭示nNOS神经元如何将可卡因线索暴露转化为复发行为的机制方面。 因此，通过防止nNOS神经元的线索激活来防止这种复发信号转导， 代表了一种潜在的治疗策略，以减少药物渴望和预防复发。

项目成果

Looking to the stars for answers: Strategies for determining how astrocytes influence neuronal activity.

• DOI: 10.1016/j.csbj.2022.07.052
• 发表时间: 2022
• 期刊: Computational and structural biotechnology journal
• 影响因子: 6

Astrocytes in Addictive Disorders.

• DOI: 10.1007/978-3-030-77375-5_10
• 发表时间: 2021
• 期刊: Advances in neurobiology

A Subset of Nucleus Accumbens Neurons Receiving Dense and Functional Prelimbic Cortical Input Are Required for Cocaine Seeking.

• DOI: 10.3389/fncel.2022.844243
• 发表时间: 2022
• 期刊: Frontiers in cellular neuroscience
• 影响因子: 5.3
• 作者: Siemsen BM;Barry SM;Vollmer KM;Green LM;Brock AG;Westphal AM;King RA;DeVries DM;Otis JM;Cowan CW;Scofield MD
• 通讯作者: Scofield MD