Emerging Activity Dynamics and Noradrenergic Modulation of Prefrontal Neuronal Ensembles During Heroin Seeking

项目摘要 阿片类药物使用障碍是一种慢性复发性疾病，其特征是强迫性药物寻求， 神经回路编排这些动机状态是未知的。这部分是由于缺乏实验性的 这是不可行的，因为难以控制和监测体内细胞类型特异性神经元的活性。为 例如，背内侧前额叶皮层（dmPFC）功能减退的发展是 物质使用障碍，可靠地预测未来复发的易感性，而重新暴露于药物相关的 提示唤起强烈的dmPFC活性。然而，dmPFC神经元的动态活动模式如何调节药物 寻找不清楚。因此，本项目的目标是使用当代工具来研究 动力学和神经回路，使dmPFC控制药物寻求。 在这里，我建议确定精确的活动动力学和去甲肾上腺素能调制的dmPFC神经元， 寻求海洛因的行为dmPFC神经元活动动力学的单细胞分辨率将使用 海洛因自我给药的头部束缚小鼠的纵向在体双光子钙成像 (Aim 1）。此外，由于在重新暴露于药物相关的药物期间观察到神经元兴奋性的快速变化， 提示可能是由去甲肾上腺素介导的，去甲肾上腺素由蓝斑（LC）在dmPFC中释放，抑制 化学遗传学将用于确定LC-dmPFC输入对提示期间活动动力学的功能， 诱导海洛因寻求（目标2）。根据我的初步数据和我的赞助商和其他人发表的工作， 我假设线索诱导的海洛因寻求性恢复直接对应于兴奋性活动 再次出现在dmPFC合奏（目的1）和LC-dmPFC激活是必要的，为线索诱导的 恢复和重现dmPFC集合中的兴奋性活动（目的2）。这些数据表明 寻求海洛因的行为是由dmPFC神经元的离散活动动力学调节的， 去甲肾上腺素能调节不管结果如何，这里提出的实验将是第一个 纵向跟踪单个神经元从吸毒开始到复发的活动。这些研究将提供 对前额叶兴奋性神经元的特性和去甲肾上腺素能机制的前所未有的了解， 海洛因寻求过程中的神经元系综动力学

PROJECT SUMMARY Opioid use disorder is a chronically relapsing disease characterized by compulsive drug seeking, but how precise neural circuits orchestrate these motivational states is unknown. This is in part due to a lack of experimental feasibility, as it has been difficult to control and monitor the activity of cell-type specific neurons in vivo. For example, development of functional hypoactivity in the dorsomedial prefrontal cortex (dmPFC) is a hallmark of substance use disorder that reliably predicts future relapse susceptibility, while re-exposure to drug-associated cues evokes robust dmPFC activity. However, how dynamic activity patterns in dmPFC neurons regulate drug seeking is unclear. Thus, the objective of this project is to use contemporary tools to study the precise activity dynamics and neurocircuitry that engages dmPFC for the control of drug seeking. Here I propose to identify the precise activity dynamics and noradrenergic modulation of dmPFC neurons for heroin-seeking behavior. Single-cell resolution of dmPFC neuronal activity dynamics will be identified using longitudinal in vivo two-photon calcium imaging in head-restrained mice undergoing heroin self-administration (Aim 1). Furthermore, as the rapid shifts in neuronal excitability observed during re-exposure to drug-associated cues may be mediated by noradrenaline, which is released in dmPFC by the locus coeruleus (LC), inhibitory chemogenetics will be used to determine the function of LC-dmPFC inputs on activity dynamics during cue- induced heroin seeking (Aim 2). Based on my preliminary data and published work by my sponsors and others, I hypothesize that cue-induced heroin-seeking reinstatement directly corresponds to excitatory activity reemergence in dmPFC ensembles (Aim 1) and that LC-dmPFC activation is necessary for both cue-induced reinstatement and reemergence of excitatory activity in dmPFC ensembles (Aim 2). These data would suggest heroin-seeking behavior is regulated by discrete activity dynamics in dmPFC neurons which are subject to noradrenergic regulation. Regardless of the results, the experiments proposed here will be the first to longitudinally track activity in single neurons from the onset of drug use to relapse. The studies will provide unprecedented insight into the characteristics and noradrenergic mechanisms of prefrontal excitatory neuronal ensemble dynamics during heroin seeking.