Making and breaking opioid memories to prevent relapse

项目摘要 反复的药物暴露产生广泛的细胞改变，可以表现为适应不良行为 和上瘾。这些行为上的改变会使人衰弱，成瘾的病理学往往是终生的 痛苦事实上，即使经过多年的禁欲，接触药物相关的药物也会加速复发。 线索这是药物成瘾的一个普遍存在的特性，并且存在于药物类别中，但我们缺乏一个明确的 了解这些变化如何能够持续如此之久。许多种类的蛋白质调节异常， 暗示诱发的复发，但复发的倾向仍然存在，远远超过这些的半衰期 蛋白质，这表明上游表观遗传变化是允许的转录景观， 会导致这些行为失常了解最终产生这些的表观遗传改变 细胞变化将大大增加可用于潜在治疗干预的靶点的数量。到 为了了解线索诱导复发的表观遗传机制，我们必须确定 这些细胞被激活来驱动寻找行为。我们的初步数据显示，在任何给定的大脑区域， 一小部分细胞对给定的刺激被激活-这组被激活的神经元被称为神经元。 合奏。因此，理解大脑的下一个前沿将是准确定义哪些细胞被激活， 什么时候被激活，为什么。在这里，我们结合联合收割机技术，使我们能够记录，操纵， 在阿片类药物自我给药和随后的线索触发的药物寻求期间序列神经系综， 确定每个神经元群体内的转录活性如何决定哪些细胞被激活。通过 在线索诱导寻找过程中结合体内细胞分辨率钙成像， 测序-在相同的动物中-我们将定义控制神经活动的转录网络 驱使人们寻找毒品的模式接下来，使用表观遗传方法和CRISPR/dCas 9融合构建体，我们 将在神经元中选择性地定义和操纵活性反应基因的表观遗传景观， 被药物相关的线索激活这个提议将使我们能够定义精确的神经集合， 寻找药物以及这些神经元内的转录网络如何控制引导药物的神经活动概况。 行为通过定义这些机制，该奖项将允许研究，推动我们如何 接近理解大脑中的信息编码，并扩展我们对如何 操纵这些过程，以减少各类毒品的复吸。

Project Summary Repeated drug exposure produces widespread cellular alterations that can manifest in maladaptive behaviors and addiction. These behavioral alterations can be debilitating, and the pathology of addiction is often a life-long affliction. Indeed, even after years of abstinence, relapse can be precipitated by exposure to drug-associated cues. This is a ubiquitous property of drug addiction, and is present across drug class, yet we lack a clear understanding of how these changes can be so long lasting. Dysregulation of many classes of proteins have been implicated in cue-evoked relapse, yet the propensity to relapse persists well past the half-life of these proteins, suggesting that upstream epigenetic changes are permissive to the transcriptional landscape that produces these behavioral aberrations. Understanding the epigenetic alterations that ultimately produce these cellular changes will great expand the number of targets available for potential therapeutic interventions. To approach understanding the epigenetic mechanisms that underlie cue-induced relapse, we must identify the cells that are activated to drive seeking behavior. Our preliminary data show that in any given brain region, only a small percentage of cells are activated to a given stimulus – this group of activated neurons is termed a neural ensemble. Thus, the next frontier of understanding the brain will be defining exactly which cells are activated, when they are activated, and why. Here we combine techniques that allow us to record, manipulate, and sequence neural ensembles during opioid self-administration and subsequent cue-triggered drug seeking to determine how transcriptional activity within each neuronal population dictates which cells are activated. By combining in vivo cellular resolution calcium imaging during cue-induced seeking followed by single cell sequencing - in the same animals - we will define the transcriptional networks that control the neural activity patterns that drive drug seeking. Next, using epigenetic approaches and CRISPR/dCas9 fusion constructs, we will define and manipulate the epigenetic landscape at activity-responsive genes selectively in neurons that are activated by drug-associated cues. This proposal will allow us to define the precise neural ensembles that guide drug seeking and how transcriptional networks within these neurons control the neural activity profiles that guide behavior. By defining these mechanisms, this award will allow research that pushes the boundaries of how we approach understanding information encoding in the brain and expand our understanding of how we can manipulate these processes to reduce relapse across drug classes.