Nucleus accumbens cholinergic interneurons and cue-induced cocaine craving

伏核胆碱能中间神经元和提示诱导的可卡因渴望

• 批准号: 10738973
• 负责人: Alexander Borg Kawa
• 金额: $ 18.53万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10738973
• 关键词: Abstinence; Advisory Committees; Affect; Affinity Chromatography; Behavior; Choline O-Acetyltransferase; Cocaine; Cocaine use disorder; Committee Members; Corpus striatum structure; Cues; Disease; Dopamine D2 Receptor; Drug Addiction; Drug usage; Enterobacteria phage P1 Cre recombinase; Exhibits; Exposure to; Female; Fiber; Genetic; Goals; Homeostasis; Human; Immunohistochemistry; Immunoprecipitation; Incentives; Incubated; Individual; Interneurons; Learning; Ligands; Link; Measures; Mediating; Memory; Mentors; Messenger RNA; Methods; Mission; Modeling; Molecular; Monitor; Motivation; Neurons; Nucleus Accumbens; Pathway interactions; Persons; Pharmaceutical Preparations; Phase; Photometry; Play; Population; Procedures; Proteins; Proxy; Quantitative Reverse Transcriptase PCR; Rat Transgene; Rattus; Receptor Signaling; Regulation; Relapse; Research; Ribosomes; Role; Saline; Self Administration; Testing; Time; Training; Translating; Translations; United States National Institutes of Health; Viral; Virus; Withdrawal; Work; addiction; biological adaptation to stress; career development; cell type; cholinergic; cocaine craving; cocaine seeking; cocaine self-administration; cocaine use; craving; designer receptors exclusively activated by designer drugs; drug abstinence; drug craving; expectation; experience; experimental study; in vivo; male; motivated behavior; neuropsychiatric disorder; novel; novel therapeutic intervention; prolonged abstinence; receptor expression; receptor function; relapse prevention; response; selective expression; skills; tool

Project Summary A major problem for persons suffering from addiction is persistent vulnerability to relapse, even after long periods of abstinence. In the `incubation of cocaine craving' model of relapse, rats self-administer cocaine using a Long Access procedure, and then experience a prolonged abstinence period. During abstinence, rats exhibit a progressive intensification (incubation) of cue-induced cocaine craving. Plasticity involving medium spiny neurons in the nucleus accumbens core (NAcc) is required for the expression of `incubated' cue-induced craving. However, to date no incubation studies have focused on another cell type in the NAcc, the Cholinergic Interneuron (CIN). CINs make up only 1-2% of NAcc neurons, but are critically involved in learning, memory, and motivated behaviors. CINs are tonically active, and considerable work now supports the idea that a reduction in their activity augments motivated behavior. Other work has shown that dopamine D2 receptors (D2-R) on CINs reduce CIN activity and this correlates with greater motivated behavior; furthermore, D2-R are upregulated in NAc following cocaine self-administration (SA). Finally, striatal CINs are unique in exhibiting steady-state activation of the Integrated Stress Response (ISR) pathway, linked to their tonic firing. The ISR maintains cellular homeostasis by regulating protein translation. In other cell types, the level of ISR activity regulates motivation for cocaine. This has not been tested for CINs, but the ISR does affect D2-R signaling in these neurons. Integrating these findings, the over-arching hypothesis to be tested here is that cocaine SA and a subsequent abstinence period leads to increased D2-R expression on NAcc CINs. This leads to a potentiated inhibition of CINs in response to DA released during cue presentation, increasing cocaine seeking and leading to reduced ISR activity and translational reprogramming in CINs. Together these changes contribute to increased motivation for cocaine-paired cues. To test this hypothesis, transgenic rats and Cre-driven viruses will be used to selectively monitor and manipulate NAcc CINs. Aim 1 will use fiber photometry to determine the effect of cocaine SA and an abstinence period on the Ca2+ response (a proxy for activity) to a cocaine-paired cue in CINs, and how D2-R regulate this response. Recordings will be performed during cue-induced seeking tests on withdrawal day (WD) 1, prior to incubation, and WD40 (after incubation). Aim 2 will determine the role that CINs play in the expression of `incubated' seeking, through bidirectionally manipulating CINs via chemogenetics prior to WD1 or WD40 seeking tests. Aim 3 will explore how incubation of cocaine craving changes protein translation in CINs, using cell type specific translating ribosome affinity purification to identify active translating mRNAs (including mRNAs for the D2-R and markers of ISR activity) and a new viral tool (SPOTlight) to measure ISR activity. While this work is underway, I will participate in a multi-faceted training plan to develop the non-bench skills needed to reach my goal of becoming an independent PI in an academic setting. I will be supported by my Mentor, co- Mentor, and other Advisory Committee members, who have complementary expertise related to my goals.

项目摘要 吸毒成瘾者的一个主要问题是，即使在很长一段时间之后， 禁欲在“可卡因渴望潜伏期”复发模型中，大鼠使用长 进入程序，然后经历一个延长的禁欲期。在禁欲期间，大鼠表现出 线索诱导的可卡因渴望的逐步强化（潜伏期）。可塑性涉及中等多刺 神经元在核神经元的核心（NAcc）是必需的表达'孵化'线索诱导的渴求。 然而，迄今为止，没有孵育研究集中在NAcc中的另一种细胞类型，胆碱能细胞， 中间神经元（CIN）。CINs仅占NAcc神经元的1-2%，但与学习，记忆， 和积极的行为。CIN是紧张性的，现在有相当多的工作支持这样的观点，即减少 在他们的活动中增强动机行为。其他研究表明，CIN上的多巴胺D2受体（D2-R） 降低CIN活性，这与更大的动机行为相关;此外，D2-R在 可卡因自我给药后的NAc（SA）。最后，纹状体CIN是唯一的表现为稳态 整合应激反应（ISR）通路的激活，与它们的紧张性放电有关。情报监视系统保持着 通过调节蛋白质翻译来维持体内平衡。在其他类型的细胞中，ISR活性水平调节细胞的生长动机。 可卡因这还没有在CIN中进行过测试，但ISR确实影响了这些神经元中的D2-R信号。整合 这些发现，这里要检验的过度兴奋假设是，可卡因SA和随后的禁欲 周期导致NAcc CIN上D2-R表达增加。这导致了对CINs的增强抑制， 对线索呈现期间释放的DA的反应，增加可卡因寻求并导致ISR活性降低 和翻译重编程。这些变化共同促进了 可卡因配对线索为了验证这一假设，转基因大鼠和Cre驱动的病毒将被用来选择性地 监控和操纵NAcc CINS。目的1将采用纤维光度法测定可卡因SA和 CINS中对可卡因配对线索的Ca 2+反应（活动的代理）的禁欲期，以及D2-R 规范这种反应。将在停药日（WD）的线索诱导寻求试验期间进行记录 1，孵育前，和WD 40（孵育后）。目的2将确定CIN在表达中所起的作用， 通过在WD 1或WD 40之前通过化学遗传学双向操纵CIN， 寻求测试。目的3将探索可卡因渴望的孵化如何改变CIN中的蛋白质翻译， 细胞类型特异性翻译核糖体亲和纯化以鉴定活性翻译mRNA（包括mRNA 用于D2-R和ISR活性标志物）和新的病毒工具（SPOTlight）来测量ISR活性。虽然这 工作正在进行中，我将参加一个多方面的培训计划，以发展所需的非板凳技能， 实现我成为学术环境中独立PI的目标。我将得到我的导师的支持，共同- 导师，和其他咨询委员会成员，谁有互补的专业知识有关我的目标。