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.

项目总结