Group I mGluR-ROS-ERK dependent sensitization in the amygdala in visceral pain

内脏疼痛中杏仁核 I 组 mGluR-ROS-ERK 依赖性敏化

基本信息

批准号：
8254402
负责人：
Volker Neugebauer
金额：
$ 22.18万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8254402
关键词：
7-(hydroxyimino)cyclopropan(b)chromen-1a-carbxoylic acid ethyl ester Abdominal Pain Action Potentials Adjuvant Therapy Adult Affective Agonist Amygdaloid structure Animals Area Arthritis Behavior Behavioral Biochemistry Brain Brain Stem Calcium Cell Nucleus Cells Chelating Agents Chemicals Clinical Colitis Collaborations Confocal Microscopy Control Groups Core Facility Corpus striatum structure Country Custom Data Disease Disease remission Electric Stimulation Electrophysiology (science)Emotional Emotions Fluorescent Dyes Goals Halothane Image Image Analysis Immunoblotting In Vitro Individual Inflammatory Bowel Diseases Injection of therapeutic agent Intracolonic Ion Channel Label Lateral Life Link MAPK1 gene MAPK3 gene MEKs Measurement Measures Mediating Membrane Metabotropic Glutamate Receptors Microdialysis Mitochondria Mitogen-Activated Protein Kinases Modeling Modification Molecular Molecular Biology Mood Disorders Neuraxis Neurons Nociception Outcome Outcome Measure Pain Pain Clinics Pain management Pathway interactions Peripheral Persistent pain Pharmaceutical Preparations Pharmacology Physiologic pulse Posterior Horn Cells Prevalence Principal Investigator Process Production Property Proteins Rattus Reactive Oxygen Species Reflex action Research Role Ryanodine Ryanodine Receptors Saline Signal Transduction Signaling Molecule Site Slice Solutions Spinal Spinal Cord Symptoms Synapses Synaptic Transmission Synaptic plasticity System Testing Thalamic structure Time Tissue Sample Tissues Translational Research U-0126 Ulcerative Colitis Ultrasonics United States Visceral pain Western Blotting Work Zymosan awake base brain tissue central sensitization clinically significant colorectal distension design dorsal column extracellular fluorescence imaging immunocytochemistry in vivo inhibitor/antagonist innovation midbrain central gray substance neurochemistry neuromechanism neuronal excitability new therapeutic target novel optical imaging oxidation pain behavior parabrachial nucleus patch clamp postsynaptic programs protein expression receptor relating to nervous system research study response therapeutic target vocalization

项目摘要

Colitis pain is clinically significant and not well understood. We hypothesize that an important mechanism in colitis pain is a novel signaling cascade in the amygdala. The amygdala, a key player in emotions and affective disorders, is emerging as an important pain center. We previously identified the latero-capsular division of the central nucleus of the amygdala (CeLC) as particularly important for pain processing and pain modulation in arthritis pain, but its role in visceral pain remains to be determined. The proposed three-step signaling cascade includes the activation of the MAP kinase ERK by group I metabotropic glutamate receptors (mGluRs) through reactive oxygen species (ROS). Our novel construct is that endogenous or exogenous activation of the mGluR->ROS->ERK signaling cascade in the amygdala contributes to visceral pain behavior and central sensitization in the spinal cord through descending pain control systems. To test this hypothesis we combine behavioral analyses, electrophysiology, pharmacology, neurochemistry and molecular biology. We use zymosan-induced colitis as a visceral pain model to determine the role of the mGluR-¿ROS-¿ERK signaling cascade in the amygdala (CeLC). The Specific Aims assess the following endpoints. 1) Visceral pain-related sensitization of CeLC neurons, using extracellular single-unit recordings in anesthetized rats before and after intracolonic zymosan or saline injection. 2) Synaptic plasticity and neural excitability of CeLC neurons, using whole-cell patch-clamp in brain slices from rats with colitis and control rats. 3) Neurochemical changes in the CeLC, using immunocytochemistry and immunoblotting of group I mGluR and ERK expression, fluorescence imaging of mitochondrial ROS, and protein oxidation measurements in tissue samples from rats with colitis and control rats. 4) Visceral pain-related behavior, including audible and ultrasonic vocalizations and visceromotor reflexes in awake rats before and after intracolonic zymosan or saline injection. 5) Central sensitization of spinal dorsal horn neurons projecting to the CeLC, using extracellular single-unit recordings before and after intracolonic zymosan or saline injection. Group I mGluR agonists and antagonists, ROS donors and scavengers, and ERK inhibitors will be administered into the CeLC by microdialysis (Specific Aims 1, 4 and 5), superfused onto brain slices (Specific Aims 2 and 3), or administered intracellularly through the patch pipette (Specific Aim 2). These studies will determine the role of the novel mGluR->ROS-¿ERK signaling cascade in the amygdala in visceral pain plasticity. This may be an important modulator of visceral pain via the amygdala. Thus, the amygdala modulates different levels of the central nervous system and contributes to the statedependent exacerbation and remission of visceral pain. This translational research will provide novel therapeutic targets for the management of visceral pain.

结肠炎疼痛具有临床意义，但我们尚未充分了解这一重要机制。结肠炎疼痛是杏仁核中的一种新型信号级联反应，杏仁核是情绪和情绪的关键参与者。情感障碍，正在成为一个重要的疼痛中心，我们之前发现了后囊。杏仁核中央核 (CeLC) 的分裂对于疼痛处理和疼痛特别重要调节关节炎疼痛，但其在内脏疼痛中的作用仍有待确定。建议分三步进行。信号级联包括 I 类代谢型谷氨酸对 MAP 激酶 ERK 的激活通过活性氧 (ROS) 受体 (mGluR) 我们的新型构建体是内源性或内源性的。杏仁核中 mGluR->ROS->ERK 信号级联的外源激活有助于内脏通过下行疼痛控制系统控制脊髓的疼痛行为和中枢敏化。为了检验这一假设，我们结合了行为分析、电生理学、药理学、神经化学我们使用酵母聚糖诱导的结肠炎作为内脏疼痛模型来确定其作用。 mGluR-¿ ROS-¿杏仁核 (CeLC) 中的 ERK 信号级联反应评估以下内容。 1）CeLC 神经元的内脏疼痛相关敏化，使用细胞外单单元记录。结肠内酵母聚糖或盐水注射之前和之后的麻醉大鼠 2) 突触可塑性和。使用全细胞膜片钳对结肠炎大鼠的脑切片进行 CeLC 神经元的神经兴奋性研究 3) 使用免疫细胞化学和免疫印迹检测 CeLC 中的神经化学变化。 I 组 mGluR 和 ERK 表达、线粒体 ROS 荧光成像和蛋白质氧化对结肠炎大鼠和对照大鼠的组织样本进行测量 4) 内脏疼痛相关行为，包括清醒大鼠前后的听觉和超声波发声以及内脏运动反射结肠内酵母聚糖或盐水注射 5) 投射到的脊髓背角神经元的中枢敏化。 CeLC，使用结肠内酵母聚糖或盐水注射前后的细胞外单单位记录。 I 组 mGluR 激动剂和拮抗剂、ROS 供体和清除剂以及 ERK 抑制剂将通过微透析（具体目标 1、4 和 5）注入 CeLC，灌注到脑切片上（具体目标 2 和 3），或通过贴片移液管进行细胞内给药（具体目标 2）。这些研究将确定新型 mGluR->ROS-¿ 的作用ERK 信号级联杏仁核在内脏疼痛可塑性中的作用，这可能是通过杏仁核调节内脏疼痛的重要因素。因此，杏仁核调节中枢神经系统的不同水平，并有助于状态依赖性这项转化研究将提供新颖的内脏疼痛的恶化和缓解。内脏疼痛管理的治疗目标。