Decoding Neuropeptide S Modulation of Reward Seeking Behavior

解码神经肽 S 对奖励寻求行为的调节

• 批准号: 10536518
• 负责人: Kasey Shea Girven
• 金额: $ 7.21万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2024-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10536518
• 关键词: Affect; Amygdaloid structure; Anatomy; Animals; Anti-Anxiety Agents; Anxiety; Anxiety Disorders; Automobile Driving; Aversive Stimulus; Behavior; Behavioral; Behavioral Assay; Biological Assay; Brain; CRISPR/Cas technology; Calcium; Cell Nucleus; Cells; Chronic stress; Coupled; Cues; Desire for food; Development; Electrophysiology (science); Female; Fiber; Food; Future; Genes; Goals; Head; Human; Hypothalamic structure; Image; In Situ Hybridization; Infusion procedures; Institution; Internal Ribosome Entry Site; Investigation; Knowledge; Lateral; Learning; Link; Measures; Molecular; Monitor; Mus; Neurobiology; Neurons; Neuropeptide Receptor; Neuropeptides; Neurosciences; Norepinephrine; Operant Conditioning; Opsin; Optics; Pharmaceutical Preparations; Pharmacology; Phase; Photometry; Play; Population; Process; Production; Property; Receptor Gene; Research; Research Personnel; Research Proposals; Rewards; Role; Series; Signal Pathway; Signal Transduction; Slice; Social Interaction; Source; Stimulus; Stress; Substance Use Disorder; Substance abuse problem; System; Techniques; Testing; Time; Training; Viral; Work; acute stress; antagonist; anxiety reduction; anxiety treatment; anxiety-related behavior; base; career; classical conditioning; comorbidity; conditioned fear; drug seeking behavior; experimental study; hindbrain; in vivo; in vivo calcium imaging; insight; interest; locus ceruleus structure; male; neuronal cell body; neuropsychiatric disorder; novel; optogenetics; parabrachial nucleus; patch clamp; receptor; response; reward processing; social; therapeutically effective; transmission process; treatment strategy; two photon microscopy; two-photon; vesicular glutamate transporter 2

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.

压力降低了动物寻求自然回报的动力，但增加了寻求药物的行为。在人类中， 焦虑症也与物质使用障碍高度共病，因此更好地了解 连接这些行为的潜在电路的研究对于开发更好的治疗方法是必要的。 战略布局研究神经肽S（NAPs）的作用，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（NPSR 1），寻求社会和自然回报的游戏可以 在身体的焦虑反应和寻求药物的行为之间提供了一个有趣的联系。基因座 蓝斑核（periLC）产生投射并将投射发送到眶额皮质（OFC），OFC含有致密的 NPSR 1的表达。使用逆行追踪与原位杂交相结合，以及 在全细胞膜片钳电生理学中，我发现OFCNPSR 1神经元接受来自periLC的输入， periLCNPS神经元投射到眶额皮层。因此，我假设periLCNPS神经元驱动自然奖励- 通过激活OFCNPSR 1神经元寻求行为。为了剖析periLCNPS-OFCNPSR 1投射， 在我的培训期间，我将学习进行新颖的体内钙成像， 网络和单细胞活动，神经药理学和分子操作。第一个目标是 研究建议是确定在自然奖赏过程中periLCNPS的内源性神经元活动， 寻求行为。先前的研究表明，LC相位活动与奖励刺激有关，使用 纤维光度法，我将测量内源性索马和终末网络活动的periLCNPS神经元 在奖励相关的行为，如经典条件反射和社会互动任务。目标1将建立 periLCNPS神经元在体内对中性和食欲刺激的行为。本研究建议的第二个目的 将研究periLCNPS-OFCNPSR 1投射对于调节是否是必要和充分的 自然的奖赏。我计划用双光子钙成像在一只行为正常的老鼠身上进行研究 在有和没有periLC末端激活的情况下，使用 一种在OFC的periLC兴奋性末梢中表达的兴奋性视蛋白。最后，我将考察 以及periLCNPS-OFCNPSR 1投射在奖赏寻求行为中的必要性。为了检验其充分性，我 将利用体内光遗传学策略来光激活经典过程中OFC中的periLC-peptide末端。 条件反射和社会互动任务。为了测试投影的必要性，我将使用一个有条件的、基于病毒的 Crispr-Cas9系统对感兴趣区域中的Nps或Npsr 1基因进行测序。这些老鼠将经历 经典条件反射和社会互动分析。在这里，我们将测试的假设，periLCNPS神经元 通过激活OFCNPSR 1神经元增强自然的奖赏寻求行为。这项工作将有助于 发展我们对periLCNPS神经元如何参与自然奖励寻求行为的理解， 揭示了对奖励相关神经精神疾病（包括药物滥用）的新见解。