Cocaine-induced mitochondrial mechanisms and molecular mediators in reward circuitry

项目摘要： 药物滥用是一种使人衰弱的复发性疾病，其特征是强迫性药物寻求和使用， 消极的个人后果。反复暴露于药物滥用伴随着持续的改变 在分子过程中，包括调节神经元可塑性改变的转录因子的改变， 功能，这是与物质使用障碍相关的持续行为反应的基础。最近 研究表明，线粒体过程可以在神经元可塑性机制中发挥作用。我们 揭示了线粒体分裂及其分子介质Drp 1在介导可塑性适应中的作用， 中脑核（NAc）表达D1的中型多棘神经元（D1-MSNs）。此外，Drp 1和裂变是 寻找可卡因的必要条件尽管有这些初步的研究，我们的研究仍然存在很大的不足。 了解线粒体过程如何整合到大脑奖励区域的电路适应中 会出现的症状此外，对上游转录因子的理解也很差， 调节介导脑奖赏区线粒体功能的分子的过程。 我们的研究将通过调查可卡因静脉自我给药的影响来弥合这一差距 （IVSA）对（1）调节MSN亚型中线粒体过程的NAc兴奋性输入及其 对药物寻求行为的影响，（2）D1-MSNs中线粒体分裂对NAc输出区的影响， （3）探讨NAc中的线粒体分子和过程及其上游转录调控因子 MSN子类型及其输入和输出区域。为了做到这一点，我们采用AAV交叉工具来操纵 或标记NAc输入和输出神经元，同时也靶向可卡因IVSA中的NAc MSN。我们使用化学基因 研究NAc兴奋性输入对线粒体动力学和分子介质影响的方法 在NAc MSN亚型中。我们进一步研究了兴奋性输入和线粒体分裂之间的相互作用 在D1-MSN上的吸毒行为。我们还研究了D1-MSNs中线粒体分裂对 在药物觅药水平下，IVSA的腹侧苍白球（VP）和腹侧被盖区（VTA）输出区 行为、多巴胺动力学和分子适应。最后，我们在整个 IVSA后奖赏回路神经元的线粒体动力学，线粒体分子介质和 转录调节因子和线粒体组学。我们进一步使用CRISPR表观基因组工具来表征可卡因- 介导的转录因子调节线粒体动力学和线粒体分子介质 奖赏回路神经元亚型。总的来说，我们的研究将为可卡因对人类的影响提供基础。 奖励回路中的线粒体过程和分子介质。

Project Summary: Drug abuse is a debilitating relapsing disease characterized by compulsive drug seeking and use despite negative personal consequences. Repeated exposure to drugs of abuse is accompanied by persistent alterations in molecular processes, including alterations in transcription factors that regulate altered neuronal plasticity and function, which underlie the persistent behavioral responses associated with substance use disorder. Recent work demonstrates that mitochondrial processes can play a role in neuronal plasticity mechanisms. We uncovered a role for mitochondria fission and its molecular mediator, Drp1, in mediating plasticity adaptations in nucleus accumbens (NAc) D1 expressing medium spiny neurons (D1-MSNs). Further, Drp1 and fission are necessary for cocaine seeking-behavior. Despite these initial studies there is still a large deficit in our understanding into how mitochondrial processes integrate into circuit adaptations across brain reward regions that occur with psychostimulant exposure. Moreover, there is poor understanding of the upstream transcriptional processes that regulate molecules mediating mitochondrial function in brain reward regions. Our studies will bridge this gap by investigating the impact of cocaine intravenous self-administration (IVSA) on (1) the excitatory inputs to NAc that regulate mitochondrial processes in MSN subtypes and their influence on drug seeking behavior, (2) the impact of mitochondrial fission in D1-MSNs on NAc output regions, and (3) explore mitochondrial molecules and processes, and their upstream transcriptional regulators, in NAc MSN subtypes and their input and output regions. To do these we employ AAV intersectional tools to manipulate or label NAc input and output neurons while also targeting NAc MSNs in cocaine IVSA. We use chemogenetic approaches to examine the impact of NAc excitatory inputs on mitochondrial dynamics and molecular mediators in NAc MSN subtypes. We further examine the interaction between excitatory inputs and mitochondrial fission in D1-MSNs on drug seeking behavior. We also examine the impact of mitochondrial fission in D1-MSNs on ventral pallidum (VP) and ventral tegmental area (VTA) output regions in IVSA at the level of drug seeking behavior, dopamine dynamics, and molecular adaptations. Finally, we perform a large scale profiling across reward circuit neurons after IVSA of mitochondrial dynamics, mitochondrial molecular mediators and transcriptional regulators, and mito-omics. We further, use CRISPR epigenome tools to characterize cocaine- mediated transcription factor regulation of mitochondrial dynamics and mitochondrial molecular mediators across reward circuit neuron subtypes. Collectively our studies will provide a foundation for the impact of cocaine on mitochondrial processes and molecular mediators in reward circuitry.