Circuit-Selective Astroglial Plasticity During Opioid Relapse

阿片类药物复发期间的电路选择性星形胶质细胞可塑性

• 批准号: 10738654
• 负责人: Anna K Kruyer
• 金额: $ 17.91万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10738654
• 关键词: ADRBK1 gene; Abstinence; Actin-Binding Protein; Animal Model; Astrocytes; Attenuated; Automobile Driving; Behavior; Behavioral; Behavioral Paradigm; Brain; Cells; Chronic; Clinical; Complement Receptor; Confocal Microscopy; Corpus striatum structure; Cues; Data; Diffusion; Dissection; Dopamine; Dopamine D1 Receptor; Drug Addiction; Drug usage; Electrophysiology (science); Epigenetic Process; Event; Exposure to; Extinction; Fiber; G protein coupled receptor kinase; Genetic Transcription; Glutamate Transporter; Glutamates; Goals; Halorhodopsins; Heroin; Homeostasis; Human; Label; Link; Literature; Measures; Mediating; Membrane; Morphology; Neurons; Nucleus Accumbens; Opioid; Opsin; Outcome; Output; Pathway interactions; Peripheral; Pharmaceutical Preparations; Phosphorylation; Photometry; Physiology; Population; Pre-Clinical Model; Process; Production; Proteins; Publishing; Rattus; Relapse; Research; Rewards; Role; Self Administration; Shapes; Signal Transduction; Sucrose; Surface; Synapses; Synaptic Transmission; Techniques; Time; Training; Transgenic Organisms; Viral; Withdrawal; Work; cell motility; cell type; drug craving; drug relapse; drug seeking behavior; effective therapy; experience; experimental study; extracellular; ezrin; glutamatergic signaling; heroin use; in vivo; insight; knock-down; link protein; motivated behavior; neural; neural circuit; neural model; neuroadaptation; neuropsychiatric disorder; neurotransmission; novel; optogenetics; postsynaptic; prolonged abstinence; protein expression; response; reward circuitry; selective expression; sensor; small hairpin RNA; tool; tool development; transmission process; uptake

Abstract Vulnerability to relapse despite prolonged abstinence is a principal feature of drug addiction. Decades of research have expanded our understanding of the neural basis of relapse, but have yielded few effective treatments that restore top-down control over drug seeking in active and former users. In animal models, repeated drug use, but not repeated exposure to natural rewards, results in excess glutamate transmission within corticofugal projections to the striatum in the presence of reward-associated cues and contexts that drive seeking. This behavioral paradigm has been used to model neural adaptations that trigger relapse, since reactivity to drug-related cues is linked to drug craving in humans. The dysregulation of glutamate transmission after chronic heroin use arises in part from enduring changes in peripheral astrocyte processes that insulate active synapses and express the glutamate transporter GLT-1, consistent with a growing literature that illustrates a prominent role for this cell type in maintaining the integrity of excitatory synaptic transmission. My published data show that astrocytes in the NAcore undergo profound transient morphological plasticity in response to drug- but not sucrose-conditioned cues and that this plasticity serves to attenuate seeking behavior. Heroin-associated cues also stimulate an increase in surface-proximity of GLT-1 on the astroglial membrane, an adaptation expected to limit glutamate spillover during relapse in preclinical models. The goal of this proposal is to explore whether measures of astroglial plasticity that I have discovered are linked to transmitter release from cortical projections within the NAcore and to uncover intracellular signaling events that trigger morphological plasticity in astrocytes during drug-seeking. A final goal of this proposal is to determine whether cue-induced astrocyte adaptations impact different post-synaptic targets, since the two principle neuronal subtypes in the NAcore, D1- and D2-MSNs differentially impact drug seeking behavior. In Aim 1, I will combine confocal microscopy with in vivo fiber photometry in order to identify the time course of astroglial plasticity during a 2-hr reinstatement session. In Aim 2, I will pair optogenetics with confocal microscopy to determine whether glutamate from prelimbic cortical terminals is necessary to drive astrocyte motility and GLT-1 surface diffusion after extinction from heroin self-administration. In Aim 3, I will interfere with the signaling cascade that drives astrocyte process motility during seeking using viral-mediated delivery of shRNAs selectively in astrocytes. Finally, in Aim 4 I will use confocal microscopy in transgenic D1- and D2-Cre rats to determine whether astrocyte processes or surface- proximal GLT-1 are selectively associated with D1- or D2-MSNs during cue-reinstated seeking. These Aims will provide me with comprehensive training in optogenetics as well as with viral tools that will allow me to selectively manipulate protein expression in astrocytes and will link my previous findings with the underlying neural circuitry. The relevance of this work extends beyond relapse to opioids and may have implications for normal synaptic physiology underlying motivated behavior.

抽象的 尽管长期戒毒，但仍容易复发是毒瘾的一个主要特征。几十年来 研究扩大了我们对复发神经基础的理解，但几乎没有取得有效的成果 恢复对活跃和前吸毒者的自上而下的药物寻求控制的治疗。在动物模型中， 重复吸毒，但不重复接触自然奖励，会导致体内谷氨酸传输过多 在存在与奖励相关的线索和背景的情况下，皮质对纹状体的投射 寻求。这种行为范式已被用来模拟触发复发的神经适应，因为 对药物相关线索的反应与人类对药物的渴望有关。谷氨酸传输失调 长期使用海洛因后，部分原因是外周星形胶质细胞过程的持久变化，这些过程隔离 活跃的突触并表达谷氨酸转运蛋白 GLT-1，这与越来越多的文献一致，这些文献表明 这种细胞类型在维持兴奋性突触传递的完整性方面发挥着重要作用。我发表的 数据显示，NAcore 中的星形胶质细胞响应药物而经历了深刻的短暂形态可塑性 但不是蔗糖条件线索，这种可塑性有助于减弱寻找行为。海洛因相关 信号还刺激星形胶质细胞膜上 GLT-1 表面接近度的增加，这是一种适应 预计将限制临床前模型复发期间谷氨酸的溢出。该提案的目标是探索 我发现的星形胶质细胞可塑性测量是否与皮质的递质释放有关 NAcore 内的投影并揭示触发形态可塑性的细胞内信号传导事件 寻求药物期间的星形胶质细胞。该提案的最终目标是确定提示诱导的星形胶质细胞是否 适应影响不同的突触后目标，因为 NAcore 中的两种主要神经元亚型 D1- 和 D2-MSN 对药物寻求行为的影响存在差异。在目标 1 中，我将把共焦显微镜与 in 体内纤维光度测定，以确定 2 小时恢复期间星形胶质细胞可塑性的时间过程 会议。在目标 2 中，我将把光遗传学与共聚焦显微镜结合起来，以确定谷氨酸是否来自 边缘前皮质末梢对于驱动星形胶质细胞运动和灭绝后 GLT-1 表面扩散是必要的 来自海洛因的自我管理。在目标 3 中，我将干扰驱动星形胶质细胞过程的信号级联 在星形胶质细胞中选择性地使用病毒介导的 shRNA 递送来寻找运动过程中的运动性。最后，在目标 4 中我将 在转基因 D1- 和 D2-Cre 大鼠中使用共聚焦显微镜来确定星形胶质细胞过程或表面- 在线索恢复寻找过程中，近端 GLT-1 选择性地与 D1-或 D2-MSN 相关。这些目标将 为我提供光遗传学方面的全面培训以及病毒工具，使我能够有选择地 操纵星形胶质细胞中的蛋白质表达，并将我之前的发现与潜在的神经回路联系起来。 这项工作的相关性不仅限于阿片类药物的复发，还可能对正常突触产生影响 动机行为背后的生理学。