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Regulation of glutamatergic signalings by niacrophage/microglia

噬菌体/小胶质细胞对谷氨酸信号的调节

基本信息

批准号：
14571764
负责人：
NAKANISHI Hiroshi
金额：
$ 1.86万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571764/
关键词：
macrophage microglia NMDA receptor electrophysiology conditioned medium 電気生理

项目摘要

To elucidate the influence of macrophage/microglia on glutamatergic synaptic transmission in the brain and bone tissues, we first examined the effects of primary cultured microglia transferred onto the organotypic cortical slice cultures. In these microglia-transferred cortical slice cultures, stimulation of the subcortlcal white matter induced fast excitatory postsynaptic potentials followed by N-merhyl-D-aspartate (NMDA) receptor-mediated plateau-like potentials that were never observed in control slice cultures. A similar potentiation of NMDA receptor-mediated postsynaptic responses was also observed by an application of a microglial-conditioned medium (MCMI 10% v/v) in acute cortical slices. These effects of MCM disappeared after boiling or incubation with proteinase K. After fractionation of MCM by anion-exchange chromatography, the enhancing activity of each fraction was quantitated electrophysiologically. When each fraction was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the fraction 24 which showed the most potent enhancing activity on NMDA receptor-mediated responses contained a relatively strong protein band with a molecular mass of approximately 70 kDa. MCM also enhanced both glutamate-and NMDA-induced inward currents recorded from acutely isolated cortical neurons. It was also noted that glutamate and NMDA induced transient large inward currents during an application of MCMI which were never observed in the control condition. These observations strongly suggest that NMDA receptor-mediated responses were potentiated by both heat-and protease-labile (presumably 70 kD-protelns) molecules released from microglia.

为了阐明巨噬细胞/小胶质细胞对脑和骨组织中神经元能突触传递的影响，我们首先研究了原代培养的小胶质细胞转移到器官型皮质切片培养物上的影响。在这些小胶质细胞转移的皮层切片培养物中，刺激皮层下白色物质诱导快速兴奋性突触后电位，随后是在对照切片培养物中从未观察到的N-甲基-D-天冬氨酸（NMDA）受体介导的平台样电位。NMDA受体介导的突触后反应也观察到类似的增强的应用程序的小胶质细胞条件培养基（MCMI 10% v/v）在急性皮质切片。这些作用在煮沸或与蛋白酶K孵育后消失。MCM经阴离子交换层析分离后，对各组分的电生理活性增强作用进行定量。当通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析每个级分时，对NMDA受体介导的反应显示出最强增强活性的级分24含有相对强的蛋白质条带，其分子量约为70 kDa。MCM还增强了谷氨酸和NMDA诱导的内向电流记录从急性分离的皮层神经元。还注意到，谷氨酸和NMDA在应用MCMI期间诱导瞬时大内向电流，这在对照条件下从未观察到。这些观察结果强烈表明，NMDA受体介导的反应是由热和蛋白酶不稳定（大概70 kD-protolns）从小胶质细胞释放的分子增强。