Isolation of large dense core vesicles to investigate the mechanisms controlling the peptidergic neurotransmitter release.

分离大的致密核心囊泡以研究控制肽能神经递质释放的机制。

• 批准号: 09672327
• 负责人: INOUE Atsuko
• 金额: $ 1.6万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672327/
• 关键词: substance P; primary afferent neuron; cultured rat spinal dorsal root ganglion cell; substance P release; nerve growth factor; preprotachykinin mRNA; interleukin-1 beta; cyclooxygenase-2; シンクロオキシゲナーゼ-2; 遊離; PPTmRNA

A bioactive undecapeptide, substance P (SP) is a neurotransmitter to convey noxious informations from peripheral tissues to the central nervous system through a primary afferent neuron. SP is bio-synthesized in a dorsal root ganglion (DRG) which is a neuronal soma of a primary afferent neuron and released from a neuronal terminal in the dorsal horn of a spinal cord by noxious stimuli. To elucidate the mechanisms controlling the release of this peptide, the effects of neurotrophins or interleukin-1beta (IL-1beta) on SP synthesis and release were investigated in the primary cultured rat DRG cells.SP is bio-synthesized by three kinds of isoforms of SP precursors, preprotachykinin (PPT) mRNA (alpha-, beta- and gamma-). Nerve growth factor (NGF) increased SP content by increased transcriptional activity of beta- and gamma- PPT mRNA in the DRG cells. IL-1beta is one of the cytokines which are synthesized and released from immune cells and considered to be important mediators during inflammation and hyperalgesia. When recombinant mouse IL-1beta was added to the DRG cells in the presence of NGF, IL-1beta evoked the SP release after 3 hours. The effect of IL-1beta ft on the SP release was Ca^<2+> dependent and significantly inhibited by a IL-1 receptor antagonist and cyclooxygenase inhibitors. Furthermore IL-1BETA increased inducible cyclooxygenase (COX)-2 mRNA without any effects on constitutive COX-1 in the incubation of 1 hour.Thus, it is suggested that IL-1beta ft evoked the release of this nociceptive neuropeptide in the DRG cells via specific IL-1 receptors, the mechanisms of which might be involved in prostanoid systems. It could be responsible for the hyperalgesic action with reference to inflammatory pain in primary afferent neuron to spinal cord pathway.

P物质(SP)是一种具有生物活性的十一肽，是一种神经递质，通过初级传入神经元将伤害性信息从外周组织传递到中枢神经系统。背根神经节(DRG)是初级传入神经元的神经元胞体，在伤害性刺激的作用下从脊髓背角的神经末梢释放出SP。为了阐明神经营养因子和白细胞介素1β(IL-1β)对SP合成和释放的影响，研究了神经营养素和IL-1β对原代培养的大鼠背根神经节细胞合成和释放SP的影响。神经生长因子(NGF)通过增加DRG细胞中β-和γ-PPT mRNA的转录活性来增加SP含量。IL-1β是由免疫细胞合成和释放的一种细胞因子，被认为是炎症和痛觉过敏过程中的重要介质。当重组小鼠IL-1β加入含有NGF的DRG细胞中时，IL-1β在3小时后可引起SP的释放。IL-1β对SP释放的影响是钙依赖性的，并可被IL-1受体拮抗剂和环氧合酶抑制剂显著抑制。此外，IL-1β作用1h可增加诱导型环氧合酶(COX)-2mRNA的表达，但对COX-1的组成成分COX-1无影响，提示IL-1β可通过特异性的IL-1受体诱导DRG细胞释放这种伤害性神经肽，其机制可能与前列腺素系统有关。它可能与初级传入神经元至脊髓通路的炎性痛有关的痛敏作用有关。

项目成果

A Inoue et al.: "Effects of neurotrophins or interleukin-1 beta on substance P synthesis in cultured rat dorsal root ganglia" Neurochemical Research. 24(1). 153 (1999)

A Inoue 等人：“神经营养素或白细胞介素 1β 对培养大鼠背根神经节 P 物质合成的影响”神经化学研究。

A Inoue et al.: "Effects of neurotrophins or interleukin-1β on substance P synthesis in cultured rat dorsal root ganglia" Neurochemical Research. 24(1). 153 (1999)

A Inoue 等人：“神经营养素或白细胞介素 1β 对培养大鼠背根神经节 P 物质合成的影响”《神经化学研究》24(1)。