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: 9753706
• 负责人: Vivian Chioma
• 金额: $ 5万
• 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/9753706
• 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.

项目总结/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基于非阿片类药物的治疗方法。 在每日使用海洛因以及使用其他滥用药物后， 已观察到髓核（NAc），其充当复发的诱发因素。这是由于 海洛因诱导的持久下调和减少星状神经胶质细胞谷氨酸转运蛋白GLT-1的摄取。 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显示， 分别减少可卡因渴望和恢复海洛因寻求的能力。恢复海洛因相关 cues受突触核核心（NAcore）中瞬时突触增强（t-SP）的调节， 由基质金属蛋白酶（MMPs）的活性介导。然而，尚不清楚NAC是否抑制MMP活性 抑制t-SP和海洛因复吸。因此，该提案概述了一系列实验， 确定NAC对t-SP的抑制和恢复的海洛因寻求是否涉及MMP活性，以及这是否是细胞类型 特定.我的初步数据表明15分钟后MMP活性降低（量化为积分密度 与生理盐水处理的大鼠相比，NAC处理的大鼠（100 mg/kg，每日一次，共5天）中线索诱导的海洛因复吸 大鼠我假设NAC在提示恢复过程中会抑制NAcore中的MMP活性，这将是 GLT-1依赖。我进一步假设NAC在D1 MSNs周围选择性抑制MMP-9活性， 恢复和增强MMP-2活性邻近D2 MSN期间灭绝。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。目的1将采用体内酶谱和GLT- 1反义吗啉策略，以评估NAC对提示海洛因复吸期间MMP活性的影响。 目的2将采用细胞特异性病毒转染NAcore中D1或D2中棘神经元（MSNs），以评估 NAC处理是否选择性地在D1或D2 MSN周围影响MMP活性， 恢复条件。除了了解MMP信号传导和NAC抑制线索恢复的能力外， 这个奖学金将训练我学习尖端技术，分析成瘾背后的突触事件。