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: 10448889
• 负责人: Anna K Kruyer
• 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: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10448889
• 关键词: ADRBK1 gene; Abstinence; Actin-Binding Protein; Address; 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 (Psychology); 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; Sum; 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 circuit; neural model; neuroadaptation; neuropsychiatric disorder; neurotransmission; novel; optogenetics; protein expression; relating to nervous system; 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 尽管节制长期是药物成瘾的主要特征，但仍有复发的脆弱性。几十年的时间 研究扩大了我们对复发神经基础的理解，但很少有效 恢复对活跃和以前用户寻求药物的自上而下控制的治疗方法。在动物模型中， 反复使用药物，但不会重复接触自然奖励，导致过多的谷氨酸传播 在有奖励相关的提示和驱动的环境的情况下，对纹状体的皮质型预测 寻求。这种行为范式已用于模拟引发复发的神经适应，因为 对与药物有关的提示的反应性与人类的渴望有关。谷氨酸传播的失调 慢性海洛因使用后，部分原因是隔离的外围星形胶质细胞过程的持久变化 主动突触并表达谷氨酸转运蛋白GLT-1，与不断增长的文献一致 该细胞类型在维持兴奋性突触传播的完整性方面的重要作用。我出版了 数据表明，纳可雷的星形胶质细胞对药物的反应经历了深刻的短暂形态可塑性。 但不是蔗糖条件的提示，并且这种可塑性可以减轻寻求行为。海洛因相关 提示还刺激了星形胶质膜上GLT-1的表面形成性的增加，适应性 预计在临床前模型中，复发期间将限制谷氨酸溢出。该提议的目的是探索 我发现的星形胶质可塑性的度量是否与从皮质释放的发射器链接有关 Nacore内的投影并发现触发形态可塑性的细胞内信号传导事件 在寻求吸毒期间的星形胶质细胞。该提案的最终目标是确定提示引起的星形胶质细胞是否 适应会影响不同的突触后靶标，因为Nacore中的两个主要神经元亚型D1- D2-MSN会差异影响寻求药物的行为。在AIM 1中，我将将共聚焦显微镜与IN结合 体内纤维光度法，以识别2小时恢复时星形胶质可塑性的时间过程 会议。在AIM 2中，我将对光遗传学与共聚焦显微镜配对，以确定是否从 前轴皮质终端是驱动星形胶质细胞运动和灭绝后GLT-1表面扩散所必需的 来自海洛因自我管理。在AIM 3中，我将干扰驱动星形胶质流程的信号级联 寻求在星形胶质细胞中选择性地使用病毒介导的shRNA递送的运动性。最后，在目标4中，我会 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是 在提示提示的寻求期间，近端GLT-1与D1-或D2-MSN有选择性相关。这些目标会 为我提供光遗传学以及病毒工具的全面培训，这些培训将使我有选择地 操纵星形胶质细胞中的蛋白质表达，并将我以前的发现与潜在的神经回路联系起来。 这项工作的相关性超出了阿片类药物的复发，可能对正常突触有影响 生理行为是动机行为的基础。