CRF-PACAP effects on anxiety circuits in mice (Bolshakov)

CRF-PACAP 对小鼠焦虑回路的影响 (Bolshakov)

• 批准号: 10116479
• 负责人: VADIM BOLSHAKOV
• 金额: $ 35.98万
• 依托单位: MCLEAN HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2024-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10116479
• 关键词: Address; Affect; Amygdaloid structure; Anxiety; Anxiety Disorders; Area; Automobile Driving; Beds; Behavior; Binding; Brain; Cell Nucleus; Cells; Clinical; Corticotropin-Releasing Hormone; Cre driver; Data; Dorsal; Education; Electrophysiology (science); Elements; Emotions; Exposure to; Fiber; Fright; Gap Junctions; Gene Expression Regulation; Glutamates; Hospitals; Human; Immobilization; Infusion procedures; Knowledge; Laboratory Animals; Lateral; Link; Mediating; Molecular; Mus; Negative Valence; Neurons; Neuropeptides; Output; Pathway interactions; Peptides; Pharmacology; Physiology; Play; Process; Production; Regulation; Reporter; Reporting; Research; Research Domain Criteria; Role; Signal Transduction; Sleep; Slice; Source; Stress; Structure; Structure of terminal stria nuclei of preoptic region; Techniques; Testing; Up-Regulation; Viral; Viral Vector; anxiety states; anxiety symptoms; anxiety-like behavior; anxiety-related behavior; base; behavior test; design; emotional behavior; experimental study; genetic approach; immunoreactivity; in vivo; interest; mouse genetics; neural circuit; new therapeutic target; optogenetics; parabrachial nucleus; pituitary adenylate cyclase activating polypeptide; pre-clinical; receptor; relating to nervous system; release factor; restraint; stressor

SUMMARY: PROJECT 2 (CRF-PACAP EFFECTS ON ANXIETYCIRCUITS IN MICE/BOLSHAKOV) Two neuropeptides, corticotropin-releasing factor (CRF) and pituitary adenylate cyclase-activating polypeptide (PACAP), have been previously implicated in the regulation of anxiety in both humans and laboratory animals. The neurocircuitry of their effects is poorly understood. Both neuropeptides are expressed in brain structures involved in control of anxiety-related states—in the basolateral amygdala (BLA) and the bed nucleus of the stria terminals (BNST), specifically—and could contribute to the production of anxiety by directing information flow in BLA-BNST circuits that regulate anxiety-related behaviors. We will address this possibility by combining the use of optogenetic techniques with ex vivo and in vivo electrophysiology, mouse genetics, tract tracing with viral vectors, and behavioral testing. In Aim 1, we will explore how the interactions between CRF- and PACAP- mediated signaling may contribute to control of signal flow in anxiety-driving BLA-BNST circuits. Optogenetically activating BLA-BNST projections, we will record from CRF-expressing neurons in different BNST subdivisions (ovBNST and adBNST) in brain slices, and explore effects of PACAP on both excitatory and inhibitory drive to recorded neurons in glutamatergic projections from BLA and their synaptically-driven spike output. Previous studies indicate that PACAP may trigger CRF release, and therefore anxiety-inducing effects of PACAP in BNST may be mediated by PACAP-induced CRF release. Thus combining pharmacological and genetic approaches, we will determine the contributions of CRFR1 and PAC1R receptors in ovBNST to the CRF-PACAP interactions that control signal flow in BLA-BNST circuits. Because the parabrachial nucleus (PBn) is the endogenous source of PACAP in BNST and CeL, in Aim 2 we will combine optogenetics in behaving mice to explore the role of CRF- PACAP interactions within BNST and lateral subnucleus of central nucleus of the amygdala (CeL), where PACAPergic fibers and CRF neurons are co-localized, in control of anxiety. Using viral tracing techniques, we will optogenetically target projections from the PBn to CRF neurons in ovBNST and/or CeL during behavioral tests that quantify anxiety-like behaviors. We will use CRF-Cre mice to ablate PAC1R receptors specifically from CRF neurons to test the possibility that activation of PAC1R specifically on ovBNST and/or CeL CRF neurons contributes to anxiety-like behavior. In Aim 3, we will explore the role of CRF-PACAP interactions in the anxiety- enhancing effects of repeated stress. We hypothesize that CRF and PACAP may act in concert to control the signal flow in anxiety-driving BLA-ovBNST-adBNST projections and that the effects of PACAP in ovBNST and/or CeL could be indirect. Project 2 may identify neural circuits that could potentially be targets for novel therapeutic treatments that can alleviate core symptoms of anxiety disorders, and as such is a key nexus of the Center that enhances, and is enhanced by, the other (preclinical, clinical) elements.

摘要：项目 2（CRF-PACAP 对小鼠/BOLSHAKOV 焦虑回路的影响） 两种神经肽，促肾上腺皮质激素释放因子（CRF）和垂体腺苷酸环化酶激活多肽 （PACAP），之前已被证明与人类和实验动物的焦虑调节有关。 人们对这两种神经肽在大脑结构中表达的神经回路知之甚少。 参与控制焦虑相关状态——基底外侧杏仁核（BLA）和纹状体床核 终端（BNST），特别是——并且可能通过引导信息流而导致焦虑的产生 调节焦虑相关行为的 BLA-BNST 回路我们将通过结合使用来解决这种可能性。 光遗传学技术与离体和体内电生理学、小鼠遗传学、病毒追踪 在目标 1 中，我们将探讨 CRF- 和 PACAP- 之间的相互作用。 介导的信号传导可能有助于控制焦虑驱动的 BLA-BNST 回路中的信号流。 激活 BLA-BNST 投影，我们将从不同 BNST 细分中表达 CRF 的神经元进行记录 （ovBNST 和 adBNST）在脑切片中，并探索 PACAP 对兴奋性和抑制性驱动的影响 记录了来自 BLA 的谷氨酸能投射的神经元及其突触驱动的尖峰输出。 研究表明 PACAP 可能触发 CRF 释放，因此 PACAP 对 BNST 产生焦虑诱导作用 可能是由 PACAP 诱导的 CRF 释放介导的，因此结合了药理学和遗传学方法， 我们将确定 ovBNST 中 CRFR1 和 PAC1R 受体对 CRF-PACAP 相互作用的贡献 因为臂旁核 (PBn) 是内源性来源。 PACAP 在 BNST 和 CeL 中的作用，在目标 2 中，我们将结合行为小鼠的光遗传学来探索 CRF-的作用 BNST 和杏仁核中央核 (CeL) 外侧亚核内的 PACAP 相互作用，其中 PACAPergic 纤维和 CRF 神经元共定位，利用病毒追踪技术控制焦虑。 将在行为过程中以光遗传学方式靶向 ovBNST 和/或 CeL 中从 PBn 到 CRF 神经元的投射 我们将使用 CRF-Cre 小鼠来专门消除 PAC1R 受体。 CRF 神经元测试 PAC1R 特异性激活 ovBNST 和/或 CeL CRF 神经元的可能性 在目标 3 中，我们将探讨 CRF-PACAP 相互作用在焦虑中的作用。 我们认为 CRF 和 PACAP 可以协同作用来控制压力。 焦虑驱动的 BLA-ovBNST-adBNST 预测中的信号流以及 PACAP 对 ovBNST 和/或 CeL 可能是间接的，项目 2 可能会识别出可能成为新型治疗目标的神经回路。 可以缓解焦虑症核心症状的治疗方法，因此是该中心的一个关键纽带 增强其他（临床前、临床）要素，并被其他要素增强。