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Imaging analysis of effect of anesthetics on intracellular calcium ion dynamics of dorsal root ganglia examined by a real-time confocal microscope

实时共聚焦显微镜观察麻醉药物对背根神经节细胞内钙离子动态影响的成像分析

基本信息

批准号：
15592127
负责人：
SATO Masahito
金额：
$ 1.92万
依托单位：
Iwate Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

Because adenosine trisphosphate (ATP) plays a pivotal role in causing inflammatory processes and pain, effects of ATP on dorsal root ganglion (DRG) has been extensively studied by electrophysiological methods. However, in the previous studies, satellite cells surrounding ganglion cells were ignored. We are skeptical about results based on isolated ganglion cells. The present study aimed to reveal the effects of ATP on ganglion and satellite cells, to this end we observed intracellular Ca^<2+> ion concentration ([Ca^<2+>]_i) change in DRG which kept structural integrities. DRG were taken out from immature (E20D) and mature (8W) rats. The [Ca^<2+>]_i changes during ATP stimulation were analysed by a confocal microscope. In immature and mature DRG, both ganglion and satellite cells responded to ATP. The ATP-induced [Ca^<2+>]_i increase of ganglion cells was rapid, and the decline was slow, while the satellite cell responses were spike-like. Only 1-3% of ganglion cells showed ATP-induced [Ca^<2+>]I changes, and about 40% of satellite cells responded. Based on the dependence of the response on presence of extracellular Ca^<2+> and the agonist, stimulations, ganglion and satellite cells possessed P2X and/or P2Y. There were no significant differences on ganglion cell responses between immature and mature DRG. In immature satellite cells, the response was single-shot, while mature satellite cells showed oscillatory fluctuation. Present results indicate that population of ganglion cells responding to ATP was low. Previous studies might confuse responses of satellite cells and those of ganglion cells. Oscillatory fluctuations appeared in mature satellite cells indicated a change of intracellular signaling mechanism during postnatal development. And this study showed that lidocaine inhibits the Ca^<2+> response of ganglion and satellite cells

由于三磷酸腺苷（ATP）在引起炎症反应和疼痛中起着关键作用，ATP对背根神经节（DRG）的作用已被广泛采用电生理方法研究。然而，在以往的研究中，卫星细胞周围的神经节细胞被忽视。我们对基于分离的神经节细胞的结果持怀疑态度。本研究观察了ATP对保持结构完整的DRG细胞内Ca^<2+>浓度（[Ca^<2+] i）的影响，旨在揭示ATP对神经节细胞和卫星细胞的作用。取未成熟（E20 D）和成熟（8 W）大鼠背根神经节。用共聚焦显微镜观察ATP刺激时细胞内[Ca^<2+>] i的变化。在未成熟和成熟的DRG中，神经节细胞和卫星细胞都对ATP有反应。ATP诱导的神经节细胞[Ca^<2+>] i增加迅速，下降缓慢，而卫星细胞反应呈尖峰状。只有1-3%的神经节细胞表现出ATP诱导的[Ca^<2+>]I变化，约40%的卫星细胞有反应。基于这种反应对细胞外Ca^<2+>和激动剂存在的依赖性，刺激、神经节细胞和卫星细胞具有P2 X和/或P2 Y。未成熟与成熟背根节神经节细胞反应无显著差异。在未成熟的卫星细胞，反应是单次射击，而成熟的卫星细胞表现出振荡波动。目前的结果表明，群体的神经节细胞响应ATP是低的。以前的研究可能会混淆卫星细胞和神经节细胞的反应。成熟的卫星细胞出现的振荡波动表明了出生后发育过程中细胞内信号机制的变化。这项研究表明利多卡因抑制神经节细胞和卫星细胞的Ca^<2+>反应