Mechanisms of pathway-specific plasticity in the incubation of craving

渴望孵化过程中路径特异性可塑性的机制

• 批准号: 10358255
• 负责人: John F Neumaier
• 金额: $ 3.48万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10358255
• 关键词: Abstinence; Affect; Animal Model; Automobile Driving; Behavior; Behavioral; Brain; Brain Diseases; Brain region; CAV2 gene; Cells; Cellular Structures; Cocaine; Cocaine Dependence; Complex; Corpus striatum structure; Data; Decision Making; Dendrites; Drug usage; Engineering; Event; Exposure to; Extinction (Psychology); Gases; Genetic Transcription; Globus Pallidus; Glutamates; Goals; Individual; Knowledge; Laboratories; LoxP-flanked allele; Measures; Mediating; Messenger RNA; Methods; MicroRNAs; Modeling; Molecular; Monitor; Neurons; Nucleus Accumbens; Output; Pathway Analysis; Pathway interactions; Pattern; Pharmaceutical Preparations; Play; Polyribosomes; Process; Protein Biosynthesis; Proteins; RNA; Rattus; Relapse; RiboTag; Ribosomes; Role; Second Messenger Systems; Self Administration; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Specific qualifier value; Synapses; Synaptic plasticity; Synaptosomes; Technology; Testing; Time; Training; Transgenes; Transgenic Organisms; Translating; Translations; Ventral Striatum; Ventral Tegmental Area; Viral Vector; addiction; adeno-associated viral vector; base; cocaine exposure; cocaine self-administration; cocaine use; combinatorial; compulsion; craving; designer receptors exclusively activated by designer drugs; drug relapse; experience; experimental study; innovation; neurotransmission; new therapeutic target; novel; novel strategies; receptor; receptor expression; response; retrograde transport; selective expression; tool; transcriptome sequencing; transgene expression; treatment strategy

Abstract Cocaine addiction involves the loss of control over drug taking so that individuals take more drug over time and can have prolonged vulnerability to relapsing to drug seeking, even after extended periods of abstinence. The progressive molecular and synaptic adaptations in neurons in the CNS that underlie these changes are not well understood, but can be studied using animal models based on extended cocaine self-administration followed by abstinence in rats. These models have shown that the nucleus accumbens core (NAcC), a small but critical brain region in ventral striatum, is implicated in compulsive cocaine taking and relapse to cocaine seeking after extended abstinence. The NAcC receives and integrates afferent information from many different brain regions and has two main output projections, the direct and indirect pathways; these pathways tend to oppose one another functional, and we predict that adaptations in signaling processes within these neurons are critical determinants affecting the relapse to drug seeking. By understanding the adaptations in cell signaling in these NAcC output neurons following extensive cocaine exposure and abstinence, we hope to contribute to novel treatment strategies for reducing the potential for relapse to drug seeking. We propose to investigate the time-dependent increase in drug seeking during abstinence known as the “incubation of craving”. We will use several innovative tools. First, we will use an intersectional viral vector approach to introduce DREADDs and other transgenic proteins to perturb and study direct and indirect pathway neurons selectively during incubation. By injecting AAV vectors with floxed and inverted transgenes into NAcC, we can activate transgene expression selectively in the direct or indirect pathway neurons by injecting the ventral tegmental area or ventral pallidum with CAV2-Cre, which is retrogradely transported to the cell bodies in NAcC. Second, we will use engineered “DREADD” receptors, a technology that we helped to establish for use in rat brain during complex behavioral experiments. DREADDs will allow us to activate Gs or Gi signaling pathways selectively in either direct or indirect pathway neurons during either repeatedly during cocaine taking or during early or late forced abstinence, thereby assessing how these canonical second messenger pathways modulate the plasticity involved in escalation or incubation. Third, we will utilize RiboTag technology to immunopurify polyribosomes selectively from direct or indirect pathway neurons and investigate the changes in RNA translation in these opposing pathways during abstinence and incubation of craving, both in cell bodies and in the synapses where activity dependent changes in local protein translation has been described. By perturbing and measuring signaling pathways in specified neurons, we hope to develop new strategies for ameliorating the adaptations associated with compulsive drug use and relapse to seeking.

抽象的 可卡因成瘾涉及对吸毒失去控制，因此随着时间的推移，个人会吸食更多的毒品，并且 即使在长期禁欲之后，也可能长期容易重新吸毒。这 这些变化背后的中枢神经系统神经元的渐进性分子和突触适应并不是 很好理解，但可以使用基于延长可卡因自我给药的动物模型进行研究 其次是大鼠的禁欲。这些模型表明伏隔核（NAcC）是一个小的 但腹侧纹状体的关键大脑区域与强迫性吸食可卡因和复发可卡因有关 寻求长期禁欲。 NAcC 接收并整合来自许多不同的传入信息 大脑区域并有两个主要的输出预测，即直接路径和间接路径；这些途径往往 功能上相互对立，我们预测这些神经元内信号传导过程的适应 是影响吸毒复发的关键决定因素。通过了解细胞的适应 在大量接触可卡因并戒断后，这些 NAcC 输出神经元中的信号传导，我们希望 有助于制定新的治疗策略，以减少吸毒复发的可能性。 我们建议调查戒酒期间寻求药物的时间依赖性增加，称为“ “贪欲的孵化”。我们将使用几种创新工具。首先，我们将使用交叉病毒载体 引入 DREADD 和其他转基因蛋白进行直接和间接干扰和研究的方法 在孵化期间选择性地通路神经元。通过注射带有 floxed 和反向转基因的 AAV 载体 进入 NAcC 后，我们可以通过以下方式在直接或间接通路神经元中选择性激活转基因表达： 将 CAV2-Cre 注射到腹侧被盖区或腹侧苍白球，该 CAV2-Cre 逆行转运至 NAcC 中的细胞体。其次，我们将使用工程化的“DREADD”受体，这是我们帮助的一项技术 建立用于复杂行为实验中大鼠大脑的使用。 DREADD 将允许我们激活 Gs 或 在重复过程中，直接或间接通路神经元中选择性地存在 Gi 信号通路 吸食可卡因或在早期或晚期强迫戒断期间，从而评估这些规范的第二个 信使途径调节涉及升级或孵化的可塑性。第三，我们将利用RiboTag 从直接或间接途径神经元中选择性地免疫纯化多核糖体并研究的技术 在戒断和渴望孵化期间，这些相反途径中 RNA 翻译的变化 在细胞体和突触中，局部蛋白质翻译的活动依赖性变化已被证实 描述的。通过扰乱和测量特定神经元的信号通路，我们希望开发新的 改善与强迫性吸毒和吸毒复发相关的适应的策略。