Peptidergic Neurotransmission in the Spinal Cord

脊髓中的肽能神经传递

• 批准号: 7082619
• 负责人: Virginia S Seybold
• 金额: $ 0.99万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-05 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7082619
• 关键词: Peptidergic; Neurotransmission; Spinal; Cord

DESCRIPTION (provided by applicant): Maintenance of hyperalgesia requires changes in the expression of proteins that lead to an increase in synaptic strength. Data generated in this proposal will test the hypothesis that peptides released in the spinal cord in response to tissue damaging stimuli activate receptors on spinal neurons that increase gene expression, resulting in increased expression of proteins that underlie hyperalgesia. We have shown that CGRP increases gene expression in spinal neurons through a CREB-dependent mechanism. We predict SP will increase protein expression through the transcription factor NFAT. Data will be generated to address the following questions: 1) Does SP activate NFAT-dependent gene transcription in spinal neurons? 2) Does SP facilitate effects of CGRP on CREB-dependent gene transcription in spinal neurons? 3) Do SP and CGRP increase expression of proteins relevant to hyperalgesia in spinal neurons in vitro? 4) Do SP and CGRP increase expression of these proteins in spinal neurons in vivo? 5) Do multiple treatments with SP and CGRP increase secondary hyperalgesia? 6) Do endogenous SP and CGRP contribute to the maintenance of secondary hyperalgesia during peripheral inflammation? Immunoblot analyses and real time PCR will be used to quantify the effects of SP and CGRP on the expression of four proteins that are known to be increased in the spinal cord during peripheral inflammation and that contribute to hyperalgesia: neurokinin-1 receptor, cyclo-oxygenase 2, type 1 nitric oxide synthase and inositol triphosphate receptor. Behavioral assays of nociception will be used to test whether multiple treatments with SP and CGRP are sufficient to increase hyperalgesia; antagonists of SP and CGRP receptors will be used to determine whether endogenous SP and CGRP are necessary to support the increase in nociceptive responses that occurs in a model of peripheral inflammation. The experiments will generate new information concerning the effects of SP and CGRP on protein expression in spinal neurons in general as well as 4 proteins that contribute to hyperalgesia. If the hypothesis is proven to be true, the data will provide evidence for a new therapeutic strategy that may be effective in treating peripheral inflammatory diseases. Because virtually all-mammalian peptides activate metabotropic receptors, the results will have broad implications for understanding the role of peptides in neurotransmission.

描述（由申请人提供）：维持痛觉过敏需要改变蛋白质的表达，从而导致突触强度的增加。该提案中生成的数据将检验以下假设：脊髓中响应组织损伤刺激而释放的肽会激活脊髓神经元上的受体，从而增加基因表达，从而导致痛觉过敏背后的蛋白质表达增加。我们已经证明 CGRP 通过 CREB ​​依赖性机制增加脊髓神经元的基因表达。我们预测 SP 将通过转录因子 NFAT 增加蛋白质表达。将生成数据来解决以下问题：1）SP 是否激活脊髓神经元中 NFAT 依赖性基因转录？ 2) SP是否促进CGRP对脊髓神经元中CREB依赖性基因转录的影响？ 3）SP和CGRP是否会增加体外脊髓神经元中与痛觉过敏相关的蛋白质的表达？ 4) SP和CGRP是否会增加体内脊髓神经元中这些蛋白质的表达？ 5) SP 和 CGRP 多次治疗是否会增加继发性痛觉过敏？ 6) 内源性 SP 和 CGRP 是否有助于维持外周炎症期间继发性痛觉过敏？免疫印迹分析和实时 PCR 将用于量化 SP 和 CGRP 对四种蛋白质表达的影响，已知这些蛋白质在外周炎症期间在脊髓中增加，并导致痛觉过敏：神经激肽-1 受体、环加氧酶2、1型一氧化氮合酶和肌醇三磷酸受体。伤害感受的行为测定将用于测试 SP 和 CGRP 的多次治疗是否足以增加痛觉过敏； SP 和 CGRP 受体拮抗剂将用于确定内源性 SP 和 CGRP 是否是支持外周炎症模型中发生的伤害性反应增加所必需的。这些实验将产生有关 SP 和 CGRP 对脊髓神经元蛋白质表达以及 4 种导致痛觉过敏的蛋白质的影响的新信息。如果该假设被证明是正确的，这些数据将为可能有效治疗外周炎症性疾病的新治疗策略提供证据。由于几乎所有哺乳动物肽都会激活代谢受体，因此这些结果将对理解肽在神经传递中的作用具有广泛的意义。