DEFINING DYNORPHIN-CRF CIRCUITS IN STRESS AND NICOTINE BEHAVIORS

描述（由申请人提供）：本研究的总体目标是更好地了解强啡肽和CRF在与尼古丁戒断相关的负面情感行为中的作用。强啡肽和CRF系统都被证明可以驱动压力和厌恶行为，但很少有研究确定这些系统如何通过扩展的杏仁核相互调节以驱动这些行为。已知强啡肽/κ阿片样物质受体（KOR）活性介导负性情绪状态诱导烦躁不安、厌恶和抑郁。CRF还通过强啡肽能相互作用产生烦躁不安、厌恶和焦虑样行为，并且已经假设CRF的释放增加可能是焦虑障碍发展的主要促成因素。因此，我们建议研究CRF和强啡肽在介导厌恶行为中的作用和相互作用，以及这是否反过来调节尼古丁戒断。这个为期五年的项目有三个具体目标。在第一个目标（在K99阶段），我们将确定是否强啡肽调节腹侧NAc的厌恶。第二个目的（在K99阶段）是研究尼古丁与强啡肽/κ阿片系统相互作用以调节负面情感行为的机制。在这里，我们将确定是否刺激强啡肽含有神经元在NAc介导的厌恶和退缩行为，这是否是KOR依赖。在这些目标中，我们将量化强啡肽释放的NAc后光遗传刺激。在R00独立阶段（目标3），我们将量化慢性尼古丁暴露前后NAc中强啡肽的释放。我们还将光遗传学刺激含有CeA-CRF的神经元并测量NAc中强啡肽的释放，以检查CRF是否调节强啡肽的释放和戒断行为特征。由于CRF和强啡肽都参与应激反应，并且初步数据表明慢性应激可以阻断KOR诱导的药物寻求，因此我们还将确定CRF1-R/KOR相互作用在暴露于应激后恢复尼古丁寻求中的作用。总之，这项工作具有重要的治疗意义，因为它将提高我们对强啡肽/CRF细胞类型的理解，调节负面情感行为的神经回路。

DESCRIPTION (provided by applicant): The overall goal of this research is to better understand role of dynorphin and CRF in negative affective behaviors that are associated with nicotine withdrawal. Both dynorphin and CRF systems have been shown to drive stress and aversive behaviors but few studies have determined how these systems modulate one another to drive these behaviors through the extended amygdala. Dynorphin/Kappa Opioid Receptor (KOR) activity has been known to mediate negative emotional states inducing, dysphoria, aversions, and depression. CRF also produces dysphoria, aversion and anxiety-like behavior via dynorphinergic interactions, and it has been hypothesized that the increased release of CRF may be a primary contributor in the development of anxiety disorders. Therefore, we propose to examine the role and interactions of CRF and dynorphin in the mediation of aversive behaviors and whether this in turn modulates nicotine withdrawal. This five-year project has three specific aims. In the first aim (during the K99 phase) we will determine whether dynorphin regulates aversion in the ventral NAc. The second aim (during the K99 phase) is to examine the mechanisms in which nicotine interacts with the dynorphin/kappa opioid system to regulate negative affective behaviors. Here we will determine whether stimulation of dynorphin containing neurons in the NAc mediates aversion and withdrawal behavior and whether this is KOR-dependent. In these aims we will quantify dynorphin release in the NAc following optogenetic stimulation. During the R00 independent phase (Aim 3) we will quantify dynorphin release in the NAc before and following chronic nicotine exposure. We will also optogenetically stimulate CeA- CRF containing neurons and measure dynorphin release in the NAc, to examine whether CRF regulates dynorphin release and behavior characteristic of withdrawal. Since, both CRF and dynorphin are involved in the stress response and preliminary data has shown that chronic stress can block KOR-induced drug seeking, we will also determine the role of CRF1-R/KOR interactions in reinstatement of nicotine seeking, following exposure to stress. Together this work has important therapeutic implications as it will enhance our understanding of dynorphin/CRF cell-types, neural circuits that modulate negative affective behaviors.