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.

项目概要 成瘾者面临的一个主要问题是即使在很长一段时间后仍然容易复发 禁欲。在复发的“可卡因渴望的孵化”模型中，老鼠使用长 进入程序，然后经历漫长的禁欲期。在禁欲期间，老鼠表现出 提示诱发的可卡因渴望逐渐增强（潜伏期）。涉及中等刺的可塑性 伏隔核核心（NAcc）中的神经元是表达“孵化”提示诱导的渴望所必需的。 然而，迄今为止，还没有孵化研究关注 NAcc 中的另一种细胞类型，即胆碱能细胞。 中间神经元（CIN）。 CIN 仅占 NAcc 神经元的 1-2%，但与学习、记忆、 和动机行为。 CIN 具有补强活性，现在大量的工作支持这样的观点：减少 在他们的活动中增强了动机行为。其他研究表明，CIN 上的多巴胺 D2 受体 (D2-R) 减少 CIN 活动，这与更大的积极行为相关；此外，D2-R 在 可卡因自我给药 (SA) 后的 NAc。最后，纹状体 CIN 在表现出稳态方面是独一无二的。 综合应激反应（ISR）通路的激活，与强直放电有关。 ISR 维持蜂窝 通过调节蛋白质翻译实现体内平衡。在其他细胞类型中，ISR 活性水平调节着 可卡因。尚未针对 CIN 进行测试，但 ISR 确实会影响这些神经元中的 D2-R 信号传导。整合 根据这些发现，这里要检验的首要假设是可卡因 SA 和随后的戒断 期导致 NAcc CIN 上 D2-R 表达增加。这导致 CI​​N 的增强抑制 对提示演示期间发布的 DA 的响应，增加可卡因寻找并导致 ISR 活动减少 以及 CIN 中的翻译重编程。这些变化共同有助于提高工作动力 可卡因配对线索。为了检验这一假设，将使用转基因大鼠和 Cre 驱动的病毒来选择性地 监控和操作 NAcc CIN。目标 1 将使用光纤光度测定法来确定可卡因 SA 和 禁欲期对 CIN 中可卡因配对线索的 Ca2+ 反应（活动的代表）的影响，以及 D2-R 如何 调节这种反应。录音将在退出日 (WD) 的提示诱导寻找测试期间进行 1，孵育前，和WD40（孵育后）。目标 2 将确定 CIN 在表达式中所扮演的角色 “孵化”寻找，通过 WD1 或 WD40 之前的化学遗传学双向操纵 CIN 寻求测试。目标 3 将探索可卡因渴望的孵化如何改变 CIN 中的蛋白质翻译，使用 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 活性标记）以及用于测量 ISR 活性的新病毒工具 (SPOTlight)。虽然这 工作正在进行中，我将参加多方面的培训计划，以培养工作所需的非工作技能 实现我在学术环境中成为独立 PI 的目标。我将得到我的导师、同事的支持 导师和其他咨询委员会成员，他们具有与我的目标相关的互补专业知识。