Progesterone and allopregnanolone of prefrontal cortical activity dynamics and heroin seeking

黄体酮和四氢孕酮对前额皮质活动动力学和海洛因寻求的影响

• 批准号: 10644613
• 负责人: Elizabeth Doncheck
• 金额: $ 19.28万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644613
• 关键词: Affect; Allopregnanolone; Animals; Behavior; Behavioral; Biological Assay; Biological Process; Cells; Clinical; Cues; Data; Development; Disease; Drug Recalls; Drug usage; Enterobacteria phage P1 Cre recombinase; Exhibits; Exposure to; Extinction; FDA approved; Female; GABA-A Receptor; Glutamates; Goals; Head; Health; Heroin; Hormonal; Hormones; Individual; Intervention; Knowledge; Label; Light; Mediating; Mentors; Mission; Monitor; Mus; Neurons; Nucleus Accumbens; Opioid; Opsin; Output; Ovarian Cycles; Ovarian hormone; Oxidoreductase; Pathway interactions; Pattern; Pharmaceutical Preparations; Phase; Predisposition; Prefrontal Cortex; Progesterone; Public Health; Publications; Regulation; Relapse; Reporting; Research; Research Personnel; Resolution; Rewards; Role; Self Administration; Sex Differences; Site; Substance Use Disorder; Testing; Time; Training; United States National Institutes of Health; analog; biological sex; calcium indicator; career; cocaine seeking; craving; drug seeking behavior; effective therapy; executive function; hormone regulation; in vivo calcium imaging; male; motivated behavior; neural; neural circuit; neurobiological mechanism; neuromechanism; neurosteroids; novel; opioid use disorder; optogenetics; positive allosteric modulator; prevent; programs; receptor; redshift; response; reward circuitry; sex; translational potential

ABSTRACT: To develop treatments, it is vital to identify neural mechanisms underlying relapse in opioid use disorder. To dissect neural activity underlying relapse, we developed a novel assay allowing for in vivo two- photon calcium imaging while head-fixed mice engage in heroin self-administration. Using this approach, we find global excitatory activity in the prelimbic-prefrontal cortex (PrL) decreases with acquisition of heroin seeking, an effect that persists through extinction, but then resurges during reinstatement. These dysregulated global PrL dynamics mirror clinical observations that are considered hallmarks of substance use disorder. Importantly, our approach has identified, for the first time, that PrL activity dynamics emerging during acquisition, extinction, and reinstatement are heterogenous, with distinct ensembles exhibiting unique excitatory and inhibitory activity dynamics aligned with behavioral epochs. How these unique activity dynamics guide drug seeking, however, is unknown. This implies greater resolution of PrL activity dynamics is necessary to determine how they functionally regulate behavior. Here, under the expert guidance of Drs. Jim Otis and Peter Kalivas, my K99 training in advanced computational neural analyses and single-cell optogenetics paired with in vivo calcium imaging will resolve the function of discrete PrL ensemble dynamics for relapse (Aim 1). Notably, there are subpopulations of individuals with substance use disorder in whom relapse may emerge through different neurobiological mechanisms. Females form one such subpopulation, as they exhibit enhanced relapse vulnerability and greater prefrontal activity during craving and relapse compared to males. However, this vulnerability covaries with the ovarian hormone cycle, such that peak circulating levels of progesterone (PROG) appear protective. These effects are mediated by its 5alpha-reductase neuroactive steroid metabolite, allopregnanolone (ALLO), which can influence prefrontal circuitry and promote adaptive responding in females. As ALLO acts as a positive allosteric modulator at GABA-A receptors, it likely constrains PrL neuronal activity to suppress drug seeking. As I find ALLO can act directly within PrL to suppress heroin-seeking reinstatement, I hypothesize that PROG and ALLO can disrupt PrL activity dynamics which functionally guide reinstatement. During the R00 phase, my career goal to be an independent investigator will involve building a research program wherein I resolve the influence of PROG and ALLO on PrL activity dynamics during reinstatement using in vivo two-photon calcium imaging. As activation of PrL projections to the nucleus accumbens core (NAcC) is necessary for reinstatement, I will retro-label PrL-NAcC neurons to enable simultaneous monitoring of global PrL and PrL-NAcC ensembles (Aim 2). Using single-cell optogenetic and circuit labelling approaches, I will assess the functional influence of PROG and ALLO on PrL and PrL-NAcC ensembles for reinstatement (Aims 2-3). As I find ALLO but not PROG suppresses reinstatement in males, I will assess steroidal effects on PrL and PrL-NAcC ensemble activity dynamics as a function of biological sex.

摘要：为了开发治疗方法，确定阿片类药物使用复发的神经机制至关重要 disorder.为了剖析复发背后的神经活动，我们开发了一种新的测定方法，允许体内两个- 光子钙成像，而头部固定的小鼠从事海洛因自我管理。使用这种方法，我们 发现边缘前额叶皮质（PrL）的整体兴奋性活动随着海洛因的获得而减少 寻求，一种通过灭绝而持续存在的效果，但在恢复过程中又重新出现。这些失调的 全球PrL动态反映了被认为是物质使用障碍标志的临床观察结果。 重要的是，我们的方法首次确定了在哺乳动物体内出现的PrL活性动态， 收购，灭绝和恢复是异质的，与不同的合奏表现出独特的 兴奋性和抑制性活动动力学与行为时期一致。这些独特的活动动力学 然而，指导药物寻求是未知的。这意味着需要更高分辨率的PrL活性动力学 来确定它们如何在功能上调节行为。在这里，在吉姆奥蒂斯博士的专家指导下， Peter Kalivas，我在高级计算神经分析和单细胞光遗传学方面的K99培训， 体内钙成像将解决复发的离散PrL集合动力学的功能（目的1）。 值得注意的是，有物质使用障碍的个体亚群，其中复发可能 通过不同的神经生物学机制出现。雌性就形成了这样一个亚群， 与男性相比，在渴望和复发期间，增强的复发脆弱性和更大的前额叶活动。 然而，这种脆弱性与卵巢激素周期相关，因此， 孕酮（PROG）似乎具有保护作用。这些作用是由其5 α-还原酶神经活性类固醇介导的 代谢产物别孕烯醇酮（allopregnanolone，ALLO），可影响前额叶回路并促进适应性反应 在女性中。由于ALLO作为GABA-A受体的正向变构调节剂，它可能抑制PrL神经元 抑制药物寻求的活动。因为我发现ALLO可以直接在PrL内部采取行动， 恢复，我假设PROG和ALLO可以破坏PrL活性动力学，功能上引导 复职在R 00阶段，我的职业目标是成为一名独立调查员，这将涉及建立一个 研究计划，其中我解决了PROG和ALLO对PrL活性动力学的影响， 使用体内双光子钙成像的恢复。由于PrL向细胞核的投射被激活， 如果PrL-NAcC神经元是恢复所必需的，我将逆向标记PrL-NAcC神经元， 同时监测全球PrL和PrL-NAcC集合（目标2）。使用单细胞光遗传学和 电路标记方法，我将评估PROG和ALLO对PrL和PrL-NAcC的功能影响 恢复合奏（目标2-3）。由于我发现ALLO而不是PROG抑制男性的恢复，我将 评估类固醇对PrL和PrL-NAcC整体活性动力学的影响，作为生物性别的函数。