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.

项目摘要： 该培训建议的主要目的是剖析基因座（LC）甲肾上腺素能的神经记录 在上下文记忆处理中，对齿状回（DG）的预测。以前的研究已经实施了 厌恶性上下文处理中的电路，去甲肾上腺素（NE）增加，导致歧视受损 在学习安全环境与不安全环境之间的区别。这些发现指向这个电路 与创伤后应激障碍（PTSD）和精神分裂症等疾病有关 以内存故障为特征，特别是在模式分离中。但是，还有更多问题 关于发生这种情况的确切电路，细胞类型和分子机制。第一个目的 培训建议旨在表征DG中去甲肾上腺素释放的空间时间动力学 厌恶上下文处理。 AIM 1A使用新开发的NE传感器结合使用光纤光度法 在厌恶和新颖的刺激期间观察内源性释放，而AIM 1B使用纤维光度法 与照片刺激的联系以测试诱发的NE释放的安全性，以模式分离代替 在中立的情况下厌恶刺激。该培训建议的第二个目标旨在阐明机制 在厌恶性的背景下，Ne对齿状回的α-和β-肾上腺素受体（ARS）作用 加工。 AIM 2a使用体外2-光子钙成像和切片电生理学，并应用AR- 确定α-和β-AR在调节DG中的特定药物的特定药物 兴奋性特性，而AIM 2B使用基于CRISPR的病毒技术来选择性敲除α-和β-ARS 在体内1-光子钙成像期间的DG中，确定α-和β-AR对DG中性的影响 合奏在厌恶性上下文处理过程中发生变化。使用这些技术将使我能够发现 α和β-ARS调节DG神经元刺激性和神经元合奏的不同机制变化为 影响厌恶的上下文处理。在这里，我们在齿状回中使用体内和体外钙成像， 齿状回中的纤维光度法，以及有选择地敲除α-和β-ARS的纤维光度法，以进一步了解 LC-DG去甲肾上腺素能及其在厌恶性上下文处理中的作用。利用这些众多 新颖的高分辨率方法将使我们能够进一步了解内源性诺雷肾上腺素系统和 确定基于PTSD和精神分裂症的潜在机制。在我量身定制的训练期间 时期，我将学习纤维光度法，2光子钙成像，体外切片电生理学，基于CRISPR 病毒，药物，计算和行为技术，并完全参与特定 梅里德开发计划为我准备独立研究职业。