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.

项目概要： 本培训计划的主要目标是解剖蓝斑（LC）去甲肾上腺素能神经回路 齿状回（DG）的投射。先前的研究表明， 厌恶性语境处理中的回路，去甲肾上腺素（NE）增加导致辨别力受损 学习安全环境和不安全环境之间的区别。这些发现表明， 与创伤后应激障碍（PTSD）和精神分裂症等疾病有关，这两种疾病都是 其特征在于存储器中的故障，特别是在模式分离中。然而，还有更多的问题 关于发生这种情况的确切电路、细胞类型和分子机制。第一个目标是 培训计划旨在描述DG中去甲肾上腺素释放的时空动态， 厌恶性语境处理Aim 1A使用光纤光度法结合新开发的NE传感器 观察在厌恶和新奇刺激过程中内源性NE的释放，而Aim 1B使用纤维光度法， 结合光刺激，以测试在模式分离中诱发NE释放的充分性， 在中性语境下的厌恶性刺激本培训建议的第二个目的是阐明 在厌恶情境中NE通过其作用于齿状回的α-和β-肾上腺素能受体（AR） 处理.目的2A使用体外双光子钙成像和切片电生理学，并应用AR- 确定α-和β-AR对特定细胞类型在调节DG中的作用的特定药理学试剂 兴奋性特性，而Aim 2B使用基于CRISPR的病毒技术选择性敲除α-和β-AR 在体内1-光子钙成像过程中DG中，以确定α-和β-AR对DG神经元的影响 在厌恶性语境处理过程中的整体转移。使用这些技术将使我能够揭示 α和β-AR调节DG神经元兴奋性和神经系综变化的不同机制， 影响厌恶性语境处理在这里，我们利用在体内和体外钙成像在齿状回， 在齿状回纤维光度测定，并选择性敲除α和β-AR，以进一步了解的作用， LC-DG去甲肾上腺素能回路及其在厌恶性语境加工中作用利用这些众多， 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能系统， 确定潜在的机制，潜在的创伤后应激障碍和精神分裂症。在我量身定制的指导培训期间， 期间，我将学习纤维光度学、双光子钙成像、体外切片电生理、CRISPR基础 病毒，药理学，计算和行为技术，以及充分参与一个特定的 指导发展计划，为我的独立研究生涯做好准备。