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.

总结：项目2（CRF-PACAP对小鼠/Bolshakov焦虑症的影响） 促肾上腺皮质激素释放因子和垂体腺苷酸环化酶激活多肽两种神经肽 （PACAP），先前已涉及人类和实验室动物的焦虑调节。 其影响的神经回路知之甚少。这两种神经肽都在大脑结构中表达 参与控制焦虑相关的状态-在基底外侧杏仁核（BLA）和床核的纹 终端（BNST），特别是-并可能有助于生产的焦虑，通过指导信息流， BLA-BNST电路调节焦虑相关行为。我们将通过结合使用 光遗传学技术与离体和体内电生理学、小鼠遗传学、用病毒的道追踪 向量和行为测试。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BLA-BNST回路中的信号流。光遗传 激活BLA-BNST投射，我们将记录不同BNST分区中表达CRF的神经元 （ovBNST和adBNST）在脑片中的作用，并探讨PACAP对兴奋性和抑制性驱动的影响， 记录来自BLA的突触能投射中的神经元及其突触驱动的尖峰输出。先前 研究表明，PACAP可能触发CRF释放，因此PACAP在BNST中具有焦虑诱导作用 可能通过PACAP诱导的CRF释放介导。因此结合药理学和遗传学方法， 我们将确定ovBNST中CRFR 1和PAC 1 R受体对CRF-PACAP相互作用的贡献 控制BLA-BNST电路中的信号流。由于臂旁核（PBn）是内源性的 在目的2中，我们将联合收割机在行为小鼠中结合光遗传学来探索CRF-1在BNST和CeL中的作用。 PACAP在BNST和杏仁中央核外侧亚核（CeL）内的相互作用，其中 PACA能纤维和CRF神经元共同定位，控制焦虑。利用病毒追踪技术，我们 将在行为过程中光遗传学靶向从PBn到ovBNST和/或CeL中CRF神经元的投射， 量化焦虑类行为的测试。我们将使用CRF-Cre小鼠特异性地消融PAC 1 R受体， CRF神经元，以测试特异性激活ovBNST和/或CeL CRF神经元上的PAC 1 R的可能性 会导致类似焦虑的行为在目标3中，我们将探讨CRF-PACAP相互作用在焦虑中的作用， 增强反复压力的效果。我们假设CRF和PACAP可能共同作用以控制 焦虑驱动BLA-ovBNST-adBNST投射中的信号流以及PACAP在ovBNST和/或 这可能是间接的。项目2可以识别可能成为新治疗药物靶点的神经回路 治疗，可以减轻焦虑症的核心症状，因此是该中心的一个关键联系， 增强其他（临床前，临床）元素，并被其他元素增强。