N-acetylcysteine Effects on Tetrapartite Synapses in Heroin Seeking

N-乙酰半胱氨酸对海洛因寻找过程中四方突触的影响

• 批准号: 9978797
• 负责人: Vivian Chioma
• 金额: $ 1.37万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2020-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978797
• 关键词: Acetylcysteine; Affect; Antioxidants; Astrocytes; Buprenorphine; Caliber; Cause of Death; Cells; Chemosensitization; Cleaved cell; Clinical Research; Clinical Trials; Cocaine; Cues; Cystine; Data; Dendritic Spines; Dopamine D1 Receptor; Dopamine D2 Receptor; Down-Regulation; Drug usage; Endopeptidases; Epidemic; Event; Exhibits; Extinction (Psychology); Extracellular Matrix; Fellowship; Fluorescein-5-isothiocyanate; Foot Process; Gelatin; Gelatinase A; Gelatinase B; Glutamate Transporter; Glutamates; Goals; Head; Heroin; Heroin Abuse; Homeostasis; Human; Individual; Injections; Knowledge; Matrix Metalloproteinases; Mediating; Methadone; Modeling; N-Methylaspartate; Neurons; Nicotine; Nucleus Accumbens; Opiate Addiction; Opioid; Outcome; Pathway interactions; Pharmaceutical Preparations; Pharmacotherapy; Population; Predisposing Factor; Process; Prodrugs; Public Health; Rattus; Relapse; Replacement Therapy; Research; Rodent; Saline; Series; Signal Transduction; Synapses; Techniques; Testing; Therapeutic; Training; Transfection; United States; Viral; Virus; addiction; base; cell type; craving; density; drug craving; drug of abuse; experience; experimental study; genetic manipulation; heroin use; in vivo; non-opioid analgesic; opioid abuse; postsynaptic; preclinical study; public health relevance; treatment effect; uptake

PROJECT SUMMARY/ABSTRACT Opioid addiction is a major public health issue. This, in part, is due to lack of non-opioid based therapeutics. Following daily heroin use, as well as use of other drugs of abuse, dysregulation in glutamate homeostasis in nucleus accumbens (NAc) has been observed, which serves as a predisposing factor to relapse. This is a result of heroin-induced enduring down-regulation and reduced uptake by the astroglial glutamate transporter, GLT-1. Clinical and preclinical studies utilizing N-acetylcysteine (NAC), an antioxidant that upregulates GLT-1, reveal its ability to reduce cocaine craving and reinstated heroin seeking, respectively. Reinstatement to heroin-associated cues is regulated by transient synaptic potentiation (t-SP) in nucleus accumbens core (NAcore), which is mediated by activity of matrix metalloproteinases (MMPs). However, it is unknown if NAC inhibits MMP activity in NAcore to inhibit t-SP and heroin reinstatement. Hence, this proposal outlines a series of experiments that will determine if NAC inhibition of t-SP and reinstated heroin seeking involves MMP activity and if this is cell-type specific. My preliminary data indicates decreased MMP activity (quantified as integrated density) after 15 minutes of cue-induced heroin reinstatement in NAC-treated rats (100 mg/kg daily X 5 days) compared to saline-treated rats. I hypothesize that NAC will suppress MMP activity in NAcore during cued-reinstatement and this will be GLT-1 dependent. I further hypothesize NAC suppresses MMP-9 activity selectively around D1 MSNs during reinstatement and enhances MMP-2 activity adjacent to D2 MSNs during extinction. These hypotheses, based upon preliminary data, will be tested through two specific aims. Aim 1 will employ in vivo zymography and GLT- 1 anti-sense morpholino strategies to assess NAC’s effects on MMP activity during cued heroin reinstatement. Aim 2 will employ cell-specific viral transfection of D1 or D2 medium spiny neurons (MSNs) in NAcore to assess whether NAC treatment effects MMP activity selectively around D1 or D2 MSNs under extinguished and reinstated conditions. In addition to understanding MMP signaling and NAC’s ability to inhibit cued reinstatement, this fellowship will train me in cutting-edge techniques for analyzing synaptic events underlying addiction.

项目概要/摘要 阿片类药物成瘾是一个重大的公共卫生问题。这在一定程度上是由于缺乏非阿片类药物的治疗方法。 每天使用海洛因以及使用其他滥用药物后，体内谷氨酸稳态失调 已观察到伏隔核（NAc），它是复发的诱发因素。这是一个结果 海洛因诱导的持久下调和星形胶质细胞谷氨酸转运蛋白 GLT-1 的摄取减少。 利用 N-乙酰半胱氨酸 (NAC)（一种上调 GLT-1 的抗氧化剂）进行的临床和临床前研究揭示了其 分别减少可卡因渴望和恢复海洛因寻求的能力。恢复与海洛因相关的活动 线索由伏隔核核心（NAcore）中的瞬时突触增强（t-SP）调节，这是 由基质金属蛋白酶（MMP）的活性介导。然而，NAC是否抑制MMP活性尚不清楚 NAcore 中抑制 t-SP 和海洛因恢复。因此，该提案概述了一系列实验，将 确定 NAC 对 t-SP 的抑制和恢复海洛因寻找是否涉及 MMP 活性以及这是否是细胞类型 具体的。我的初步数据表明 15 分钟后 MMP 活性下降（量化为积分密度） 与盐水处理的大鼠相比，NAC 处理的大鼠（每日 100 毫克/千克 X 5 天）提示诱导的海洛因恢复的效果 老鼠。我假设 NAC 将在提示恢复期间抑制 NAcore 中的 MMP 活性，这将是 GLT-1 依赖性。我进一步假设 NAC 在 D1 MSN 周围选择性抑制 MMP-9 活性 在灭绝期间恢复并增强与 D2 MSN 相邻的 MMP-2 活性。这些假设，基于 根据初步数据，将通过两个具体目标进行测试。目标 1 将采用体内酶谱法和 GLT- 1 反义吗啉策略评估 NAC 在提示海洛因恢复期间对 MMP 活性的影响。 目标 2 将采用 NAcore 中 D1 或 D2 中型多棘神经元 (MSN) 的细胞特异性病毒转染来评估 NAC处理是否选择性地影响D1或D2 MSN周围的MMP活性 恢复了条件。除了了解 MMP 信号传导和 NAC 抑制提示恢复的能力之外， 这项奖学金将培训我分析成瘾背后的突触事件的尖端技术。