Amygdala pain mechanisms

杏仁核疼痛机制

• 批准号: 10390809
• 负责人: Volker Neugebauer
• 金额: $ 55.16万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-15 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10390809
• 关键词: Acute; Acute Pain; Address; Affective; Amygdaloid structure; Anxiety; Area; Astrocytes; Basic Science; Behavior; Behavioral; Behavioral Assay; Binding; Bioinformatics; Brain; Cell Nucleus; Cells; Chronic; Clinical; Complex; Corticotropin-Releasing Hormone; Development; Disease; Electrophysiology (science); Elements; Emotional; Emotions; Female; Funding; Genes; Healthcare; Immunohistochemistry; Individual; Interdisciplinary Study; Knowledge; Label; Lateral; Ligation; Measures; Membrane; Mental Depression; Microglia; Modeling; Molecular; Molecular Biology; Molecular Profiling; Molecular Target; Neuraxis; Neuroglia; Neuroimmune; Neuronal Plasticity; Neurons; Nociception; Outcome Measure; Output; Pain; Pain management; Pathway interactions; Persistent pain; Pharmaceutical Preparations; Pharmacology; Phase; Physiology; Play; Pontine structure; Property; Rattus; Regulation; Research Project Grants; Resolution; Role; Sensory; Sensory Thresholds; Signal Transduction; Slice; Spinal; Spinal nerve structure; Spine pain; Sucrose; Synapses; Synaptic Transmission; System; Testing; Therapeutic; Time; Transgenic Organisms; United States National Institutes of Health; Viral; Viral Vector; Work; base; cell type; chronic neuropathic pain; chronic pain; forced swim test; improved; innovation; insight; interdisciplinary approach; male; molecular drug target; molecular marker; novel; optogenetics; pain behavior; pain chronification; pain model; pain relief; painful neuropathy; patch clamp; peripheral pain; preference; protein expression; public health relevance; response; sexual dimorphism; single-cell RNA sequencing; spontaneous pain; tool; transcriptomics; vocalization

Project Summary Chronic pain, a complex multidimensional disorder, remains a major health care issue and a therapeutic challenge. The development of new and improved therapeutic strategies requires the full understanding of mechanisms of chronic pain at all levels of the neuraxis and for all cell types, including non-neuronal elements. Neuroimmune signaling has emerged as a peripheral and spinal pain mechanism, but little is known about the role, regulation and therapeutic potential of molecular crosstalk between neuronal and glial cell types in the brain in the context of pain. To address this important knowledge gap we will build on our NIH-funded work (since 1999) that impacted the field by identifying neuroplasticity in the amygdala, a brain center for emotions, as a critical mechanism for emotional-affective aspects of pain and pain modulation. The proposed project will test the novel hypothesis that chronification of amygdala plasticity and neuropathic pain behaviors depends on a cascade of neuroimmune signaling that can be targeted to mitigate neuropathic pain. Specifically, enhanced synaptic drive of certain amygdala neuron types at the acute pain stage activates different types of glia, and glia-derived factors generate hyperexcitability in distinct amygdala neuron types at the chronic stage to maintain neuropathic pain. A comprehensive multidisciplinary approach will be used that integrates state-of-the-art transcriptomics, bioinformatics, behavioral assays, brain slice electrophysiology, pharmacology, chemogenetics, optogenetics, viral vector strategies, immunohistochemistry and molecular biology for the analysis of neuroimmune interactions within and between different types of neuronal and glial cells in the amygdala output region (central nucleus, CeA) in the well-established spinal nerve ligation (SNL) rat model of neuropathic pain. Male and female rats will be studied. A transgenic Crh-Cre rat model will be used to study the CeA corticotropin releasing factor (CRF) system, an important player in amygdala plasticity, and its role in neuroimmune signaling. Aim 1 will use a combination of cell-specific bulk and single-cell RNA sequencing (scRNA-Seq) to identify individual genes, molecular pathways and functional cellular states associated with pain chronification. A novel computational scRNA-Seq- based interactome analysis will determine neuron-glia interactions at the acute and chronic stages of neuropathic pain and identify drug-targetable molecular factors. Aim 2 will determine the behavioral significance of neuron-glia-neuron signaling in the CeA at different stages of neuropathic pain, using chemogenetic activation and inhibition of different cell types and pharmacological (or viral based) tools to modulate molecular factors. Sensory thresholds, emotional responses, non-evoked ongoing pain, and anxiety- and depression-like behaviors will be measured. Aim 3 will determine electrophysiological mechanisms of neuron-glia-neuron signaling in the CeA at different stages of neuropathic pain, using patch-clamp recordings of different cell types in brain slices and chemogenetic and pharmacological (or viral based) manipulations. Successful completion of these conceptually innovative studies will significantly advance our knowledge of neuro-immune signaling in brain plasticity and in the transition from acute to chronic pain and provide novel targets to mitigate chronic neuropathic pain.

项目摘要 慢性疼痛是一种复杂的多维障碍，仍然是一个主要的医疗保健问题和治疗性挑战。这 开发新的和改进的治疗策略需要对慢性疼痛机制充分理解 在所有级别的神经级别和所有细胞类型（包括非神经元元素）上。神经免疫信号已经出现 作为一种外围和脊柱疼痛机制，但对的作用，调节和治疗潜力知之甚少 在疼痛的背景下，大脑中神经元和神经胶质细胞类型之间的分子串扰。解决这个重要的 知识差距我们将基于NIH资助的工作（自1999年以来），该工作通过识别神经可塑性影响了该领域 在杏仁核中，一个脑部情绪中心，是疼痛和疼痛情绪影响方面的关键机制 调制。拟议的项目将检验一个新的假设，即杏仁核可塑性和 神经性疼痛行为取决于一系列可用于减轻的神经免疫信号传导 神经性疼痛。具体而言，急性疼痛阶段某些杏仁核神经元类型的突触驱动器增强 激活不同类型的神经胶质，而胶质衍生的因素在不同的杏仁核类型中产生过度刺激性 维持神经性疼痛的慢性阶段。将使用一种全面的多学科方法 整合最先进的转录组学，生物信息学，行为测定，脑切片电生理学， 药理学，化学遗传学，光遗传学，病毒载体策略，免疫组织化学和分子生物学 杏仁核中不同类型的神经元和神经胶质细胞内和之间的神经免疫性相互作用的分析 神经性疼痛的公认脊柱神经结扎术（SNL）大鼠模型中的输出区域（中央核，CEA）。 将研究雄性和雌性大鼠。转基因CRH-CRE大鼠模型将用于研究CEA Corticotropin 释放因子（CRF）系统，杏仁核可塑性的重要参与者及其在神经免疫信号传导中的作用。目标1 将使用细胞特异性大量和单细胞RNA测序（SCRNA-SEQ）的组合来识别单个基因， 分子途径和功能性细胞状态与疼痛变子有关。一种新颖的计算scrna-seq- 基于相互作用的分析将在神经性疼痛的急性和慢性阶段确定神经元-GLIA相互作用 并确定可靶向药物的分子因素。 AIM 2将确定神经元-Glia-Neuron的行为意义 使用化学发生激活和抑制不同细胞的神经性疼痛的不同阶段的CEA信号传导 调节分子因素的类型和药理（或基于病毒的）工具。感官阈值，情感 将测量反应，持续不断的疼痛以及焦虑和抑郁症的行为。 AIM 3将确定 在神经性疼痛的不同阶段，神经元 - 神经元信号传导的电生理机制， 使用脑切片中不同细胞类型的贴片钳记录以及化学生物和药理（或病毒 基于）操纵。这些概念上创新研究的成功完成将大大推动我们的 了解脑可塑性中神经免疫信号传导以及从急性到慢性疼痛的过渡 减轻慢性神经性疼痛的新颖靶标。