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: 9318063
• 负责人: John F Neumaier
• 金额: $ 42.65万
• 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 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9318063
• 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; Injectable; 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; 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 use; combinatorial; compulsion; craving; designer receptors exclusively activated by designer drugs; drug relapse; experience; experimental study; innovation; intersectionality; 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.

摘要 吸毒成瘾涉及对吸毒的失控，使个人随着时间的推移服用更多的药物， 即使在长时间的禁欲之后，也可能有长期的复吸倾向。的 中枢神经系统中神经元的渐进性分子和突触适应是这些变化的基础， 很好理解，但可以使用基于长期可卡因自我给药的动物模型进行研究 随后在大鼠中进行禁欲。这些模型已经表明，在核内，一个小的 但腹侧纹状体的关键脑区与可卡因的强迫性服用和复发有关 寻求延长禁欲期NAcC接收和整合来自许多不同的传入信息。 大脑区域，有两个主要的输出投射，直接和间接途径;这些途径往往 相反，另一个功能，我们预测，适应信号过程中，这些神经元 是影响重新吸毒的关键决定因素。通过了解细胞的适应性 在这些NAcC输出神经元中的信号传导在广泛的可卡因暴露和戒断后，我们希望 有助于新的治疗策略，以减少潜在的复发药物寻求。 我们建议调查戒毒期间寻求药物的时间依赖性增加， “渴望的孵化”。我们将使用一些创新的工具。首先，我们将使用一个交叉病毒载体 引入DREADDs和其他转基因蛋白直接和间接干扰和研究的方法 在孵育期间选择性地传递神经元。通过注射带有floxed和inverted转基因的AAV载体， 进入NAcC，我们可以激活转基因表达选择性地在直接或间接途径神经元， 用CAV 2-Cre注射腹侧被盖区或腹侧苍白球，CAV 2-Cre逆行运输到 NAcC中的细胞体。第二，我们将使用工程化的“DREADD”受体，这是一项我们帮助 建立用于复杂行为实验中的大鼠大脑。DREADDs将允许我们激活Gs或 Gi信号通路选择性地在直接或间接通路神经元中重复进行， 可卡因服用或在早期或晚期被迫禁欲，从而评估如何这些典型的第二次 信使途径调节参与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免疫纯化多聚核糖体技术，并研究 在戒断和渴望的孵化过程中，这些相反的通路中RNA翻译的变化， 在细胞体和突触中，局部蛋白质翻译的活性依赖性变化已经被 介绍了通过干扰和测量特定神经元中的信号通路，我们希望开发新的 改善与强迫性药物使用和寻求复发相关的适应性的策略。