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背后的神经机制至关重要 紊乱。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，允许体内两种 光子钙成像，同时头部固定的小鼠进行海洛因自我给药。使用这种方法，我们 发现前边缘前额皮质 (PrL) 的整体兴奋性活动随着海洛因的获取而减少 寻求，这种效应在灭绝过程中持续存在，但在恢复过程中又重新出现。这些失调的 全球 PrL 动态反映了被视为物质使用障碍标志的临床观察结果。 重要的是，我们的方法首次发现，PrL 活动动态在 获得、灭绝和恢复是异质的，不同的集合表现出独特的特征 兴奋性和抑制性活动动态与行为时期一致。这些独特的活动动态如何 然而，指导寻药却是未知的。这意味着需要更高分辨率的 PrL 活动动态 以确定它们如何在功能上调节行为。在这里，在Drs的专家指导下。吉姆·奥蒂斯和 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 PrL 系综动力学对于复发的功能（目标 1）。 值得注意的是，有一些患有物质使用障碍的个体亚群可能会复发 通过不同的神经生物学机制出现。女性就是这样一个亚群，正如她们所表现的那样 与男性相比，在渴望和复吸期间，复吸的脆弱性增强，前额叶活动更大。 然而，这种脆弱性与卵巢激素周期共变，因此峰值循环水平 黄体酮 (PROG) 具有保护作用。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)，可以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 受体的正变构调节剂，它可能限制 PrL 神经元 抑制寻求毒品的活动。我发现 ALLO 可以直接在 PrL 内起作用来抑制海洛因寻求 恢复后，我假设 PROG 和 ALLO 可以破坏 PrL 活动动态，而 PrL 活动动态在功能上引导 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 活动动态的影响 使用体内双光子钙成像进行恢复。随着 PrL 向细胞核投射的激活 伏隔核 (NAcC) 对于恢复是必要的，我将重新标记 PrL-NAcC 神经元以启用 同时监测全球 PrL 和 PrL-NAcC 系综（目标 2）。利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 恢复乐团（目标 2-3）。因为我发现 ALLO 而不是 PROG 会抑制男性的恢复，所以我会 评估类固醇对 PrL 和 PrL-NAcC 整体活动动力学的影响，作为生物性别的函数。