Neurotrophins and Neuropeptides in Colon and Bladder Hypersensitivity

结肠和膀胱过敏中的神经营养素和神经肽

• 批准号: 8439022
• 负责人: Liya Qiao
• 金额: $ 33.14万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8439022
• 关键词: Address; Afferent Neurons; Behavioral; Biological; Bladder; Bladder Control; Brain-Derived Neurotrophic Factor; Calcitonin Gene-Related Peptide; Calcium; Clinical Trials; Colitis; Colon; Complex; Cyclic AMP-Responsive DNA-Binding Protein; Data; Dominant-Negative Mutation; Down-Regulation; Drug Delivery Systems; Electrophysiology (science); Frequencies; Goals; Human; Hyperactive behavior; Hypersensitivity; Immune Sera; In Vitro; Inflammation; Inflammatory Bowel Diseases; Intracolonic; Knowledge; Label; Measurement; Mediating; Mediator of activation protein; Messenger RNA; Molecular; Molecular Profiling; Neurogenic Bladder; Neurons; Neuropeptides; Pain; Pathway interactions; Patients; Phospholipase C; Play; Post-Translational Regulation; Preparation; Protein Kinase; Proteins; Publications; Rat Strains; Rattus; Regulation; Research Project Grants; Role; Sensory; Sensory Physiology; Series; Signal Pathway; Signal Transduction; Spinal Ganglia; Sulfonic Acids; Symptoms; System; Testing; Therapeutic; Transcriptional Regulation; Transfection; Transgenic Animals; Up-Regulation; Urination; Visceral; Work; behavior test; calmodulin-dependent protein kinase II; experience; extracellular; in vivo; inhibitor/antagonist; insight; neurochemistry; neurotrophic factor; neutralizing antibody; nitrobenzene; phospholipase C gamma; protein activation; public health relevance; receptor; small molecule; voltage

DESCRIPTION (provided by applicant): Patients with inflammatory bowel disease often experience increased sensory responsiveness in the urinary bladder reflecting neurogenic bladder overactivity. This visceral symptom overlap involves neuronal cross-activation in the dorsal root ganglia (DRG) where the bladder afferent neurons are sensitized. The long- term goal of this project has been to understand the complex neuronal interaction between colonic and bladder sensory pathways, and to identify mediators that regulate bladder sensory hypersensitivity as a result of colitis. Our publications and preliminary data have shown that the brain-derived neurotrophic factor (BDNF)/TrkB system has a prominent role in the regulation of bladder activity. In this renewal application, we hypothesize that the phospholipase C-gamma (PLC?)-calcium (Ca2+) pathways are unique downstream of the increased endogenous BDNF/TrkB in bladder afferent neurons, and play an integral role in bladder afferent activation by Ca2+-dependent transcriptional and posttranslational regulation of neuroactive compounds during colitis. To address this hypothesis, three interrelated Specific Aims are proposed to examine the regulatory mechanism and the targets of the PLC?-Ca2+ pathways including Ca2+/calmodulin-dependent protein kinase (CaMK)II, cAMP-response element binding protein (CREB), calcitonin gene-related peptide (CGRP), and cerebellin 1 precursor (Cbln1) in bladder afferent neurons before and during colitis. In AIM 1, we will characterize the expression profiles of a series of components regulating Ca2+ mobilization (phospholipase C?, InsP3R-1, voltage-gated Ca2+ channels predominantly the N-type channel Cav2.2) and Ca2+-dependent neuronal activation (CaMKII and CREB) in bladder afferent neurons at 7 days and 21 days of colitis. This will be done at the molecular (mRNA and protein) and functional (intracellular Ca2+ recording and electrophysiology) levels. In AIM 2, we will examine the regulatory mechanism by which the PLC?-Ca2+ pathways are activated by endogenous BDNF in bladder afferent neurons during colitis. For this purpose, we will use a newly developed yet well-characterized BDNF+/- rat strain. In AIM 3, we will combine molecular biological, pharmacological, neurochemical, and behavioral tests to examine the functional role of the BDNF-Ca2+ axis in bladder hyperactivity during colitis. We will characterize how the Ca2+-dependent pathways are involved in CGRP and Cbln1 expression, and how they regulate bladder afferent neuronal hyperactivity and modulate bladder micturition parameters during colitis. For studies proposed above, we will utilize a variety of in vivo (transgenic animals, intrathecal delivery of drugs, behavioral studies and ex vivo/in vitro (DRG explants, isolated DRG neuron culture and transfection) systems. The localized colonic inflammation will be induced by intracolonic instillation of tri-nitrobenzene sulfonic acid (TNBS) in rat. As several small molecule antagonists of the Ca2+ pathways are under clinical trials in treatment of other pain symptoms, we anticipate that the current systematic studies will provide insights into forming therapeutic strategies in the treatment of visceral hypersensitivity.

描述（由申请人提供）：患有炎症性肠病的患者经常会经历膀胱感觉反应性增加，反映神经源性膀胱过度活动。这种内脏症状重叠涉及背根神经节（DRG）中的神经元交叉激活，其中膀胱传入神经元变得敏感。该项目的长期目标是了解结肠和膀胱感觉通路之间复杂的神经元相互作用，并确定调节结肠炎引起的膀胱感觉超敏反应的介质。我们的出版物和初步数据表明，脑源性神经营养因子（BDNF）/TrkB 系统在膀胱活动的调节中具有显着的作用。在此更新应用中，我们假设磷脂酶 C-γ (PLC?)-钙 (Ca2+) 途径是膀胱传入神经元中内源性 BDNF/TrkB 增加的独特下游，并且在 Ca2+- 的膀胱传入激活中发挥着不可或缺的作用。结肠炎期间神经活性化合物的依赖性转录和翻译后调节。为了解决这一假设，提出了三个相互关联的具体目标来检查 PLC?-Ca2+ 途径的调节机制和靶标，包括 Ca2+/钙调蛋白依赖性蛋白激酶 (CaMK)II、cAMP 反应元件结合蛋白 (CREB)、降钙素结肠炎之前和期间膀胱传入神经元中的基因相关肽（CGRP）和小脑蛋白1前体（Cbln1）。在 AIM 1 中，我们将描述一系列调节 Ca2+ 动员的成分（磷脂酶 C?、InsP3R-1、电压门控 Ca2+ 通道（主要是 N 型通道 Cav2.2））和 Ca2+ 依赖性神经元激活（CaMKII）的表达谱。和 CREB）在结肠炎 7 天和 21 天的膀胱传入神经元中。这将在分子（mRNA 和蛋白质）和功能（细胞内 Ca2+ 记录和电生理学）水平上完成。在 AIM 2 中，我们将研究结肠炎期间膀胱传入神经元中内源性 BDNF 激活 PLC?-Ca2+ 通路的调节机制。为此，我们将使用新开发的但特征良好的 BDNF+/- 大鼠品系。在 AIM 3 中，我们将结合分子生物学、药理学、神经化学和行为测试来检查 BDNF-Ca2+ 轴在结肠炎期间膀胱过度活动中的功能作用。我们将描述 Ca2+ 依赖性途径如何参与 CGRP 和 Cbln1 表达，以及它们如何在结肠炎期间调节膀胱传入神经元过度活跃和调节膀胱排尿参数。对于上述提出的研究，我们将利用各种体内（转基因动物、鞘内给药、行为研究和离体/体外（DRG 外植体、分离的 DRG 神经元培养和转染）系统。将诱导局部结肠炎症由于Ca2+途径的几种小分子拮抗剂正在接受治疗其他疼痛症状的临床试验，我们预计当前的系统研究将通过向大鼠结肠内滴注三硝基苯磺酸（TNBS）来实现。为形成内脏过敏症的治疗策略提供见解。