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

• 批准号: 7496225
• 负责人: Volker Neugebauer
• 金额: $ 21.17万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 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/7496225
• 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; Biochemistry and Pharmacology Cancer Activity; Brain; Brain Stem; Calcium; Cell Nucleus; Cells; Chelating Agents; Chemicals; Chromosome Pairing; Clinical; Colitis; Collaborations; Confocal Microscopy; Control Groups; Core Facility; Corpus striatum structure; Country; Custom; Data; Disease; Disease remission; Electric Stimulation; Electromyography; Electrophysiology (science); Emotional; Emotions; End Point; Extracellular Signal Regulated Kinases; Fluorescent Dyes; Ganciclovir; Glutamate Receptor; 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; Persistent pain; Pharmaceutical Preparations; Pharmacology; Physiologic pulse; Placement; Posterior Horn Cells; Prevalence; Principal Investigator; Process; Production; Property; Proteins; Pulse taking; Rattus; Reactive Oxygen Species; Reflex action; Research; Role; Ryanodine; Ryanodine Receptor Calcium Release Channel; Saline; Signal Transduction; Signaling Molecule; Site; Slice; Solutions; Spinal; Spinal Cord; Symptoms; Synapses; Synaptic Transmission; Synaptic plasticity; System; Testing; Tetrodotoxin; 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; concept; design; dihydroxyphenylethylene glycol; dorsal column; extracellular; fluorescence imaging; immunocytochemistry; in vivo; inhibitor/antagonist; innovation; midbrain central gray substance; neurochemistry; neuromechanism; neuronal excitability; novel; novel therapeutics; optical imaging; oxidation; parabrachial nucleus; patch clamp; postsynaptic; programs; protein expression; receptor; relating to nervous system; research study; response; therapeutic target; vocalization

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，尚不清楚。我们假设这是一个重要的机制 结肠炎疼痛是杏仁核中新型的信号级联。杏仁核，情绪的关键人物， 情感障碍，正在成为重要的疼痛中心。我们先前确定了latero-caplar 杏仁核（CELC）中央核的划分对于疼痛处理和疼痛特别重要 关节炎疼痛的调节，但其在内脏疼痛中的作用仍有待确定。提议的三步 信号传导级联反应包括I组的MAP激酶ERK激活ERK 通过活性氧（ROS）的受体（mglurs）。我们的新颖结构是内源性或 杏仁核中mglur-> ros-> erk信号级联的外源激活有助于内脏 通过下降的疼痛控制系统，脊髓中的疼痛行为和中枢灵敏度。 为了检验该假设，我们结合了行为分析，电生理学，药理学，神经化学 和分子生物学。我们使用Zymosan诱导的结肠炎作为内脏疼痛模型来确定 杏仁核（CELC）中的mglur-€ros-erk信号级联。具体目的评估以下 端点。 1）使用细胞外单单元记录，CELC神经元的内脏疼痛相关敏感性 在麻醉大鼠中，在殖民地内和盐水注射剂之前和之后。 2）突触可塑性和 CELC神经元的神经刺激性，使用大鼠的大脑切片中的全细胞贴剂钳，患有结肠炎和 控制大鼠。 3）使用免疫细胞化学和免疫印迹的CELC的神经化学变化 I组MGlur和ERK表达，线粒体ROS的荧光成像以及蛋白质氧化 来自结肠炎和对照大鼠大鼠的组织样品的测量。 4）内脏疼痛相关的行为， 包括听觉和超声波发声以及在清醒大鼠之前和之后的内脏反射 殖民地Zymosan或盐水注射。 5）脊柱背角神经元的中心灵敏度投射到 CELC，使用细胞外单单元记录前后注射盐含量之前和之后。 I组mglur激动剂和对手，ROS捐献者和清除剂以及ERK抑制剂将是 通过微透析（特定目的1、4和5）在CELC中施用，超级为脑切片 （特定目标2和3），或通过贴片移液管内细胞内给药（特定的目标2）。 这些研究将确定新型mglur-> ros-€erk信号级联的作用 内脏疼痛可塑性中的杏仁核。这可能是通过杏仁核的内脏疼痛的重要调节剂。 这，杏仁核调节了中枢神经系统的不同水平，并有助于陈述的 恶化和缓解内脏疼痛。这项翻译研究将提供小说 治疗靶标的内脏疼痛的靶标。