Decoding Neuropeptide S Modulation of Reward Seeking Behavior

压力降低了动物寻求自然回报的动力，但增加了寻求毒品的行为。在人类身上， 焦虑症也与物质使用障碍高度并存，因此更好地理解 对于开发更好的治疗方法来说，连接这些行为的底层电路是必要的 战略。研究神经肽S的作用，神经肽是一种调节焦虑相关行为的神经肽 通过激活GQ偶联蛋白受体(NPSR1)，在寻求社会和自然回报方面发挥作用可以 在身体的焦虑反应和寻求毒品行为之间提供了一个有趣的联系。周缘轨迹 蓝斑(PeriLc)产生NPS，并向包含致密物质的眶前叶皮质(OFC)发出投射 NPSR1的表达。使用逆行追踪和原位杂交相结合的方法，以及 全细胞膜片钳电生理学，我发现OFCNPSR1神经元接受来自周围LC的输入，并且 PERLCNPS神经元投射到OFC。因此，我假设周围LCNPS神经元驱动自然奖赏- 通过激活OFCNPSR1神经元寻找行为。为了解剖PeriLCNPS-OFCNPSR1投影 在奖赏行为中的作用，在我的训练期间，我会学习进行新颖的体内钙成像 网络和单细胞活性、神经药理学和分子操纵。这样做的第一个目的是 研究建议确定在自然奖赏过程中PeriLCNPS的内源性神经元活动。 寻找行为。先前的研究表明，LC时相活动与奖励刺激有关，使用 纤维光度法，我将测量周围LCNPS神经元的内源性胞体和终末网络活动 在与奖励相关的行为中，如经典的条件反射和社会互动任务。目标1将建立 PERI LCNPS神经元在体内对中性和食欲刺激的行为。这项研究提案的第二个目的是 将调查PeriLCNPS-OFCNPSR1投影对于调制是否必要和充分 自然而然地寻求回报。我计划在一只行为正常的老鼠身上使用双光子钙成像技术来研究 OFCNPSR1在有和没有周围LC末端激活的食欲刺激传递过程中的单细胞活性 OFC内LC周围兴奋性终末表达的兴奋性视蛋白。最后，我将检查其充分性 以及PERI-LCNPS-OFCNPSR1投射在奖赏行为中的必要性。为了检查充分性，我 将利用在体光发生策略来光激活OFC中的周围LC-NPS终末 条件反射和社交任务。为了测试预测的必要性，我将使用有条件的、基于病毒的 CRISPR-Cas9系统使感兴趣区域的Nps或Npsr1基因失活。然后这些小鼠就会经历 经典的条件反射和社会互动分析。在这里，我们将检验LCNPS神经元周围的假设 通过激活OFCNPSR1神经元增强自然的奖赏行为。这项工作将有助于 发展我们对PeriLCNPS神经元如何参与自然的奖赏寻求行为和 揭示了对包括药物滥用在内的与奖励相关的神经精神疾病的NPS的新见解。

Stress reduces an animal’s drive for seeking natural rewards but increases drug-seeking behavior. In humans, anxiety disorders are also highly comorbid with substance-use disorder, and therefore a better understanding of the underlying circuitry that connect these behaviors is necessary for the development of better treatment strategies. Examining the role neuropeptide S (NPS), a neuropeptide shown to regulate anxiety-related behavior through activation of the Gq-coupled protein receptor (NPSR1), plays in seeking social and natural rewards could provide an interesting link between the body’s anxiety response with drug-seeking behaviors. The peri locus coeruleus (periLC) produces NPS and sends projections to the orbitofrontal cortex (OFC), which contains dense expression of NPSR1. Using a combination of retrograde tracing paired with in situ hybridization, as well as whole-cell patch clamp electrophysiology, I found that the OFCNPSR1 neurons receive input from the periLC, and that periLCNPS neurons project to the OFC. Therefore, I hypothesize that periLCNPS neurons drive natural reward- seeking behavior through activation of OFCNPSR1 neurons. In order to dissect the periLCNPS-OFCNPSR1 projection’s role in reward-seeking behavior, during my training period, I will learn to conduct novel in vivo calcium imaging of network and single-cell activity, neuropharmacological and molecular manipulations. The first aim of this research proposal is to determine the endogenous neuronal activity of periLCNPS during natural reward- seeking behaviors. Previous research indicates that LC phasic activity is associated with reward stimuli, using fiber photometry, I will measure the endogenous soma and terminal network activity of the periLCNPS neurons during reward-related behaviors such as classical conditioning and social interaction tasks. Aim 1 will establish how periLCNPS neurons behave in vivo to neutral and appetitive stimuli. The second aim of this research proposal will investigate whether periLCNPS-OFCNPSR1 projections are necessary and sufficient for the modulation of natural reward-seeking. I plan to use two-photon calcium imaging in a behaving mouse to investigate OFCNPSR1 single-cell activity during delivery of appetitive stimuli with and without periLC terminal activation using an excitatory opsin expressed in periLC excitatory terminals in the OFC. Finally, I will examine the sufficiency and necessity of the periLCNPS-OFCNPSR1 projection in reward-seeking behaviors. To examine the sufficiency, I will utilize in vivo optogenetic strategies to photoactivate the periLC-NPS terminals in the OFC during classical conditioning and social interaction tasks. To test the projection’s necessity, I will use a conditional, viral-based Crispr-Cas9 system to inactivate Nps or Npsr1 genes in the region of interest. These mice will then undergo classical conditioning and social interaction assays. Here we will test the hypothesis that periLCNPS neurons enhance natural reward-seeking behavior through activation of OFCNPSR1 neurons. This work will serve to develop our understanding of how periLCNPS neurons are involved in natural reward-seeking behaviors and reveal new insights into NPS in reward-related neuropsychiatric diseases including substance abuse.