Role of trpv4 in trigeminally mediated nociception

trpv4 在三叉神经介导的伤害感受中的作用

基本信息

批准号：
8069929
负责人：
WOLFGANG B. LIEDTKE
金额：
$ 45.86万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8069929
关键词：
Action Potentials Address Afferent Neurons Animal Model Animals Bacterial Infections Behavior Behavior Control Behavioral Behavioral Assay Brain Stem Calcium Caliber Capsaicin Cell Nucleus Cells Chemicals Chromosomes Clinical Collaborations Dependence Dura Mater Electrophysiology (science)Enterobacteria phage P1 Cre recombinase Environment Event Face G-Protein-Coupled Receptors Gene Expression Generations Genes Genetic Genotype Human Resources Hyperalgesia Image Inflammation Inflammation Mediators Investigation Ion Channel Ions Knock-out Knockout Mice Label Lead Life Light Mechanics Mediating Modality Molecular Mus Neuroanatomy Neurons Nociception Nociceptors PAR-2 Receptor Paclitaxel Pain Pain Disorder Pathway interactions Peripheral Play Principal Investigator Process Proteins Public Health Rattus Relative (related person)Reporter Reporter Genes Research Personnel Role Sensory Signal Pathway Signal Transduction Sodium Sodium Channel Stimulus Structure Structure of trigeminal ganglion Structure of trigeminal nerve spinal tract nucleus Sunburn TRPV1 gene Tamoxifen Testing Transgenic Mice Transgenic Organisms Trigeminal Neuralgia Trigeminal Nuclei Trigeminal System Trigeminal nerve structure Universities Ursidae Family Vanilloid base capsaicin receptor epithelial Na+ channel in vivo insight loss of function mouse model nerve supply neuron development neuronal cell body nociceptive response novel patch clamp programs promoter receptor research study response tissue/cell culture translational neuroscience transmission process voltage voltage gated channel

项目摘要

DESCRIPTION (provided by applicant): Trigeminally-mediated pain disorders are incompletely understood at the molecular level. TRP(V) ion channels have been implicated to function in trigeminal sensory transduction. TRPV4 is multimodally activated, e.g. by mechanical, thermal and tonicity stimuli, and products of the trpv4 gene have been detected in trigeminal ganglion sensory neurons and their peripheral projections. trpv4-/-mice, previously generated by the PI, have an elevated threshold for noxious mechanical and tonicity stimuli. The critical question remains unresolved whether and how TRPV4+ trigeminal sensory neurons transduce such stimuli that elicit nociceptive behavior. Thus, the objective of this proposal is to investigate molecular and cellular mechanisms of frpv4-mediated transduction of noxious osmotic and mechanical stimuli in trigeminal sensory neurons by development of a novel mouse model. The hypothesis to be tested is whether trpv4 gene expression in trigeminal sensory neurons is critical for neurosensory transduction in response to noxious stimuli. Specifically, (1) primary cultures of trigeminal neurons from trpv4-/- mice will be investigated by patch-clamp and Ca++ imaging in response to tonicity and mechanical stimuli, modulated by activation of proteinase-activated receptor 2 (PAR-2), a physiologically relevant pro-algesic pathway, and for their "tonicity-tuning" of voltage-gated sodium channels (lNa) and capsaicin-response; (2) same as in Aim (1), but for dissociated sensory neurons from sensory-neuron-specific, inducible trpv4-/- mice; (3) sensdry-neuronspecific, inducible trpv4-/- mice will be generated to test the response of these mice to noxious osmotic stimuli and osmotic modulation of capsaicin stimuli of trigeminal peripherals, modulated by activation of PAR-2. For Aims (1)-(2), primary cultured neurons will be labeled by a genetically-encoded fluorescent reporter gene, expressed in a bacterialTartificial chromosome transgenic line of mice. For Aim (3), mice will be subjected to nocifensive behavioral assays and c-FOS studies of the brain stem trigeminal spinal nucleus in response to stimulation. Thus, the dependence of sensory transduction and transmission of noxious trigeminal stimuli on trpv4 will be deconstructed at the cellular level, clarifying the relative contribution of the trigeminal sensory neuron. Moreover, light will be shed on how trpv4 functions in "tonicity-tuning" of the capsaicin response and of INa-. and on modulation of TRPV4 by PAR-2. Thus, our new mouse model will help us better understand how trigeminal ganglion neurons respond to noxious stimuli and which role the trpv4 gene plays in this process, hopefully opening up new avenues for treatment of trigeminally-mediated pain.

描述（由申请人提供）：在分子水平上未完全理解三角介导的疼痛障碍。 TRP（V）离子通道已与三叉剂感觉转导的功能有关。 TRPV4是多模式激活的，例如在三叉神经节感官神经元及其周围投影中，通过机械，热和良性刺激以及TRPV4基因的产物。以前由PI生成的TRPV4 - / - 小鼠具有有害的机械和良性刺激的阈值升高。关键的问题仍未解决，是否以及如何转导TRPV4+三叉神经元会引起伤害性行为。因此，该提案的目的是通过开发新的小鼠模型来研究三叉神经感觉神经元中有害渗透和机械刺激的分子和细胞机制。要检验的假设是，三叉学感觉神经元中的TRPV4基因表达是否对于响应有害刺激的神经感觉转导至关重要。 Specifically, (1) primary cultures of trigeminal neurons from trpv4-/- mice will be investigated by patch-clamp and Ca++ imaging in response to tonicity and mechanical stimuli, modulated by activation of proteinase-activated receptor 2 (PAR-2), a physiologically relevant pro-algesic pathway, and for their "tonicity-tuning" of voltage-gated sodium channels (lNa) and辣椒素反应；（2）与AIM（1）相同，但对于来自感觉 - 神经元特异性的，诱导的TRPV4 - / - 小鼠的分离的感觉神经元；（3）将生成感官 - 神经特异性的TRPV4 - / - 小鼠，以测试这些小鼠对三叉神经外围周可以的辣椒素刺激的有害渗透刺激和渗透调节的反应，该刺激是通过PAR-2的激活来调节的。对于目标（1） - （2），原代培养的神经元将被遗传编码的荧光报告基因标记，该基因在小鼠的细菌型染色体染色体转基因系中表达。对于AIM（3），小鼠将对脑干三叉神经脊髓核的副行为分析和C-FOS研究，以响应刺激。因此，有害三叉神经刺激对TRPV4的感觉转导和传播的依赖性将在细胞水平上解构，从而阐明三叉神经感觉神经元的相对贡献。此外，关于辣椒素反应和ina-的“调节性调节”的功能如何在trpv4起作用。以及通过PAR-2对TRPV4的调制。因此，我们的新小鼠模型将帮助我们更好地了解三叉神经节神经元如何对有害刺激做出反应以及TRPV4基因在此过程中起哪个作用，希望能为三角形介导的疼痛打开新的途径。