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: 10579997
• 负责人: VADIM BOLSHAKOV
• 金额: $ 35.97万
• 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 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579997
• 关键词: Ablation; 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; Experimental Designs; Exposure to; Fiber; Fright; 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; 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; behavior test; emotional behavior; genetic approach; immunoreactivity; in vivo; interest; mouse genetics; neural; neural circuit; new therapeutic target; optogenetics; parabrachial nucleus; pharmacologic; pituitary adenylate cyclase activating polypeptide; pre-clinical; receptor; release factor; restraint; stressor; synergism

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/布尔沙科夫焦虑症的影响） 两种神经肽，皮质激素释放因子（CRF）和倍酰胺腺苷酸环化酶激活多肽 （PACAP）以前在人类和实验动物的动画调节中已实施。 其作用的神经循环尚未理解。两种神经肽在大脑结构中均表达 参与与动画相关状态的控制 - 基础杏仁核（BLA）和纹状体的床核 特别是终端（BNST），并且可以通过指导信息流来促进动画的生产 调节与动画相关的行为的BLA-BNST电路。我们将通过结合使用来解决这种可能性 带有体内和体内电生理学，小鼠遗传学的光遗传学技术，病毒式追踪 向量和行为测试。在AIM 1中，我们将探讨CRF-和PACAP-之间的相互作用如何 介导的信号传导可能有助于控制焦虑驱动的BLA-BNST电路中的信号流。光遗传学 激活BLA-BNST项目，我们将从不同BNST细分的表达CRF的神经元中记录 （OVBNST和ADBNST）在大脑切片中，并探索PACAP对兴奋性和抑制性驱动的影响 来自BLA的谷氨酸能项目及其突触驱动的尖峰输出记录了神经元。以前的 研究表明，PACAP可能会触发CRF释放，因此焦虑引起的PACAP在BNST中的影响 可以通过PACAP诱导的CRF释放介导。结合了药物和遗传方法， 我们将确定OVBNST中CRFR1和PAC1R受体对CRF-PACAP相互作用的贡献 BLA-BNST电路中的控制信号流。因为伞菌核（PBN）是内源性源 在BNST和CEL中的PACAP，在AIM 2中，我们将在行为小鼠中结合光遗传学以探索CRF​​-的作用 杏仁核（CEL）的BNST和外侧核中的PACAP相互作用，其中 远下纤维和CRF神经元共定位在动画中。使用病毒跟踪技术，我们 在行为期间，将在OVBNST和/或CEL中从PBN到CRF神经元的光源性项目。 测试可以量化焦虑般的行为。我们将使用CRF-CRE小鼠专门从 CRF神经元测试PAC1R在OVBNST和/或CEL CRF神经元上的激活的可能性 有助于类似焦虑的行为。在AIM 3中，我们将探讨CRF-PACAP相互作用在焦虑中的作用 - 增强重复应力的影响。我们假设CRF和PACAP可能会共同行动以控制 焦虑驱动的bla-ovbnst-adbnst项目中的信号流以及PACAP在OVBNST和/或 CEL可能是间接的。项目2可以识别有可能成为新疗法目标的神经回路 可以减轻动画障碍的核心症状的治疗方法，因此是中心的关键联系 另一个（临床前，临床）元素增强并增强了。