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：要开发治疗，至关重要的是确定阿片类药物使用中基本复发的神经机制 紊乱。为了剖析复发基础的神经活动，我们开发了一种新颖的测定法，允许在体内二 - 当头固定小鼠的光子钙成像从事海洛因自我给药。使用这种方法，我们 在预先获得海洛因时，查找前额外皮层（PRL）中的全球兴奋活动降低 寻求，这种效果通过灭绝而持续存在，但随后在恢复原状过程中重新恢复。这些失调 全球PRL动力学反映了被认为是药物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PRL活动动态。 采集，灭绝和恢复是异质的，具有独特的独特的集合 与行为时期一致的兴奋性和抑制活性动力学。这些独特的活动动态如何 但是，寻求药物是未知的。这意味着需要更大的PRL活性动力分辨率 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（Jim Otis）和 彼得·卡利瓦斯（Peter Kalivas），我在高级计算神经分析和单细胞光遗传学配对的K99培训 使用体内钙成像将解析离散PRL集成动力学的功能以进行复发（AIM 1）。 值得注意的是，有一些患有药物使用障碍的人的亚群，复发可能 通过不同的神经生物学机制出现。女性形成这样的亚群，它们表现出来 与男性相比，渴望和复发期间的复发脆弱性和更大的前额叶活性增强。 但是，这种脆弱性与卵巢激素周期的协变量，使得峰值循环水平 孕酮（Prog）看起来有保护。这些作用是由其5alpha-还原酶神经活性类固醇介导的 代谢产物，异戊烯醇（Allo），可以影响前额叶电路并促进适应性反应 在女性中。由于Allo在GABA-A受体上充当阳性变构调节剂，因此可能会限制PRL神经元 活动以抑制吸毒。正如我发现Allo可以直接在PRL中采取行动以抑制寻求海洛因 恢复原状，我假设Prog和Allo可以破坏PRL活动动态，从而在功能上指导 复职。在R00阶段，我成为独立调查员的职业目标将涉及建造一个 研究计划，我解决了Prog和Allo对PRL活动动态的影响 使用体内两光子钙成像恢复原状。随着PRL投影向细胞核的激活 伏隔核（NACC）对于恢复原状是必要的，我将重新标记PRL-NACC神经元启用 同时监视全球PRL和PRL-NACC合奏（AIM 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 恢复的合奏（目标2-3）。正如我发现Allo的那样，但没有抑制男性的恢复原状，我会 评估对PRL和PRL-NACC集成活性动力学动态的类固醇影响，这是生物学的函数。