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中复发的神经机制至关重要。 无序。为了剖析复发背后的神经活动，我们开发了一种新的检测方法，允许在体内进行两种- 头部固定小鼠进行海洛因自我给药时的光子钙成像。使用这种方法，我们 前额叶皮质(PRL)的整体兴奋性活动随着海洛因的获得而降低 寻求，一种在灭绝过程中持续存在的效果，但在恢复时又会复活。这些不受监管的 全球PRL动力学反映了被认为是物质使用障碍特征的临床观察。 重要的是，我们的方法第一次确定了PRL活动的动态 获取、灭绝和恢复是异质性的，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代相一致。这些独特的活动动态是如何 然而，指导找药是未知的。这意味着PRL活动动态的更大分辨率是必要的 以确定它们如何在功能上规范行为。在这里，在吉姆·奥蒂斯博士和 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 体内钙成像将解决离散的PRL系综动力学对复发的作用(目标1)。 值得注意的是，有一些患有药物使用障碍的人可能会复发。 通过不同的神经生物学机制出现。雌性就是这样的一个亚群，就像她们展示的那样 与男性相比，在渴望和复发期间，复发的易感性和更大的前额叶活动。 然而，这种脆弱性与卵巢激素周期有关，因此循环中的峰值水平 黄体酮(PROG)似乎具有保护性。这些作用是由其5α-还原酶神经活性类固醇介导的。 代谢物，别孕酮(ALLO)，它可以影响前额叶电路，促进适应性反应 在雌性身上。由于Allo在GABA-A受体上起到正变构调节剂的作用，它可能会抑制PRL神经元 打压毒品寻人的活动。因为我发现Allo可以在PRL内部直接采取行动来镇压海洛因寻觅 恢复，我假设prog和allo可以扰乱PRL活动的动态，它在功能上引导 复职。在R00阶段，我成为一名独立调查员的职业目标将包括建立一个 研究计划，其中我解决了Prog和Allo对PRL活性动态的影响 利用体内双光子钙成像进行复原术。因为PRL向细胞核的投射被激活 伏隔核(NACC)是恢复所必需的，我将追溯标记PRL-NACC神经元以使能 同时监测全球PRL和PRL-NACC总体(目标2)。利用单细胞光生和 电路标记方法，我将评估Prog和Allo对PRL和PRL-NAcc的功能影响 复职合奏(目标2-3)。当我发现Allo而不是Prog抑制男性的复职时，我会 作为生物性别的函数，评估类固醇对PRL和PRL-NACC系综活性动态的影响。