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）：炎症性肠病患者通常会在反映神经源性膀胱过度活动的尿道膀胱中会增加感觉反应性。这种内脏的症状重叠涉及在背根神经节（DRG）中神经元的交叉激活，其中膀胱传入神经元敏感。该项目的长期目标是了解结肠和膀胱感觉途径之间复杂的神经元相互作用，并确定调节因结肠炎而调节膀胱感觉超敏反应的介体。我们的出版物和初步数据表明，脑衍生的神经营养因子（BDNF）/TRKB系统在调节膀胱活性中具有重要作用。 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结肠炎之前和期间传入神经元。在AIM 1中，我们将表征一系列调节Ca2+动员的组件的表达谱（磷脂酶C？，INSP3R-1，电压门控Ca2+通道主要是N型通道CAV2.2）和Ca2+ ca2+依赖性神经元激活（CAMKKII和CREB）在Bladder neurons and Neurons and Neurons and Atterent and arterent and arterent and arterents and arterent and arterent and arterent and arter。这将在分子（mRNA和蛋白质）以及功能（细胞内CA2+记录和电生理学）水平上完成。在AIM 2中，我们将检查CLC+途径在结肠炎过程中膀胱传统神经元中的内源性BDNF激活的调节机制。为此，我们将使用新开发但特征良好的BDNF +/-大鼠菌株。在AIM 3中，我们将结合分子生物学，药理，神经化学和行为测试，以检查BDNF-CA2+轴在结肠炎期间膀胱多动中的功能作用。我们将表征CA2+依赖性途径如何参与CGRP和CBLN1表达，以及它们如何调节膀胱传入神经元多动并调节结肠炎期间的膀胱排尿参数。对于上面提出的研究，我们将利用各种体内（转基因动物，药物鞘内输送，行为研究和外体外/体外（DRG外植体，孤立的DRG神经元培养和转染）系统。局部结肠炎症将由脑内的多种含量（Tri-Nentrobenzene sulection intrip and Indins Indic）诱发，构成了多种含量（Tn硝基含量）。 CA2+途径正在治疗其他疼痛症状的临床试验下，我们预计当前的系统研究将为您在治疗内脏超敏反应时提供有关形成治疗策略的见解。