Elucidating the Role of Noradrenergic Circuitry in Aversive Contextual Processing

阐明去甲肾上腺素能回路在厌恶语境处理中的作用

• 批准号: 10604128
• 负责人: Eric Zhang
• 金额: $ 4.68万
• 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/10604128
• 关键词: Acute; Adrenergic Agents; Adrenergic Antagonists; Adrenergic Receptor; Adult; Affect; Agonist; Animals; Anxiety; Arousal; Automobile Driving; Aversive Stimulus; Behavioral; Biosensor; Brain; Brain Stem; Brain region; Calcium; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Development Plans; Discrimination; Disease; Electrophysiology (science); Event; Failure; Fiber; Frequencies; Future; Glutamates; Goals; Hippocampus (Brain); Image; Impairment; In Vitro; International; Interneurons; Knock-out; Learning; Lifting; Literature; Mediating; Memory; Mental disorders; Mentors; Modernization; Molecular; National Institute of Mental Health; Neurobiology; Neurons; Neurosciences; Norepinephrine; Odors; Pattern; Persons; Pharmacology; Photometry; Photons; Play; Population; Post-Traumatic Stress Disorders; Prevalence; Process; Property; Publishing; Receptor Signaling; Regulation; Research; Research Personnel; Resolution; Role; Schizophrenia; Shock; Signal Transduction; Slice; Source; Stimulus; Stress; Structure; Synaptic plasticity; System; Tail; Techniques; Technology; Testing; Time; Training; Viral; Virus; Work; antagonist; base; behavioral outcome; beta-2 Adrenergic Receptors; beta-adrenergic receptor; career; cell type; dentate gyrus; experience; foot; hippocampal subregions; in vivo; locus ceruleus structure; memory process; neural circuit; neurobehavioral; neuroregulation; noradrenergic; norepinephrine system; novel; postsynaptic; relating to nervous system; response; sensor; skills; spatiotemporal; stressor; two-photon

Project Summary: The primary goal of this training proposal is to dissect the neurocircuitry of locus coeruleus (LC) noradrenergic projections to the dentate gyrus (DG) in contextual memory processing. Previous research has implicated this circuit in aversive contextual processing, with increased norepinephrine (NE) resulting in impaired discrimination in learning the difference between a safe context and an unsafe context. These findings point to this circuits relevance to disorders such as post-traumatic stress disorder (PTSD), and schizophrenia, both of which are characterized by failures in memory, specifically in pattern separation. However, many more questions remain regarding the exact circuit, cell-type, and molecular mechanisms by which this occurs. The first aim of this training proposal seeks to characterize the spatiotemporal dynamics of norepinephrine release in the DG during aversive contextual processing. Aim 1A uses fiber photometry in conjunction with newly developed NE-sensors to observe endogenous NE release during aversive and novel stimuli, while Aim 1B uses fiber photometry in conjunction with photo-stimulation to test for sufficiency of evoked NE release in pattern separation in place of aversive stimuli in a neutral context. The second aim of this training proposal seeks to elucidate the mechanisms by which NE acts on α- and β-adrenergic receptors (ARs) in the dentate gyrus during aversive contextual processing. Aim 2A uses in vitro 2-photon calcium imaging and slice electrophysiology with application of AR- specific pharmacological agents to determine the role of α- and β-ARs on specific cell types in modulating DG excitability properties, while Aim 2B uses CRISPR-based viral techniques to selectively knockout α- and β-ARs in the DG during in vivo 1-photon calcium imaging to determine the effects of α- and β-ARs on DG neural ensemble shifts during aversive contextual processing. Using these techniques will allow me to uncover the distinct mechanisms by which α and β-ARs modulate DG neuron excitability and neural ensemble changes to affect aversive contextual processing. Here we utilize in vivo and in vitro calcium imaging in the dentate gyrus, fiber photometry in the dentate gyrus, and selectively knockout α- and β-ARs to further understand the role of the LC-DG noradrenergic circuit and its role in aversive contextual processing. Leveraging these numerous, novel, high-resolution approaches will allow us to further understand the endogenous noradrenergic system and identify potential mechanisms that underlie PTSD and schizophrenia. During my tailored mentored training period, I will learn fiber photometry, 2-photon calcium imaging, in vitro slice electrophysiology, CRISPR-based viral, pharmacological, computational, and behavioral techniques, as well as fully engage into a specific mentored development plan to prepare me for an independent research career.

项目总结： 本训练计划的主要目的是解剖蓝斑去甲肾上腺素能神经回路。 语境记忆加工中向齿状回(DG)的投射。此前的研究已经证明了这一点 厌恶背景加工中的回路，去甲肾上腺素(NE)增加导致辨别能力受损 学习安全环境和不安全环境之间的区别。这些发现指向了这个回路 与创伤后应激障碍(PTSD)和精神分裂症等疾病的相关性，这两者都是 以内存故障为特征，特别是在模式分离方面。然而，还有更多的问题存在。 关于这种情况发生的确切电路、细胞类型和分子机制。这样做的第一个目的是 训练计划试图描述DG在DG中释放去甲肾上腺素的时空动力学。 厌恶的语境处理。Aim 1A将光纤光度测量与新开发的NE传感器结合使用 观察厌恶刺激和新刺激时内源性去甲肾上腺素的释放，而Aim 1B使用纤维光度法在 联合光刺激检测模式分离中诱发NE释放的充分性 中性环境中的厌恶刺激。本培训方案的第二个目的是试图阐明这些机制。 在厌恶情境中NE对齿状回α和β肾上腺素能受体的作用 正在处理。AIM 2A使用体外双光子钙成像和切片电生理学，并应用AR- 确定特定细胞类型上的α-和β-ARs在调节DG中的作用的特定药物 兴奋性特性，而Aim 2B使用基于CRISPR的病毒技术选择性地敲除α-和β-AR 在体单光子钙显像法测定α-和β-Ars对DG神经的影响 在厌恶的语境加工过程中，集合转移。使用这些技术将使我能够发现 α和β-AR调节DG神经元兴奋性和神经丛变化的不同机制 影响令人厌恶的语境处理。在这里，我们利用体内和体外的齿状回钙成像， 纤维光度法在齿状回，并选择性地敲除α-和β-ARs，以进一步了解 LC-DG去甲肾上腺素能回路及其在厌恶情境加工中的作用。利用这些众多的、 新的、高分辨率的方法将使我们能够进一步了解内源性去甲肾上腺素系统和 找出创伤后应激障碍和精神分裂症的潜在机制。在我量身定做的指导培训期间 期间，我将学习纤维光度学、双光子钙成像、体外切片电生理学、CRISPR为主 病毒、药理学、计算和行为技术，以及完全参与特定的 指导我的发展计划，为我的独立研究生涯做准备。