神经细胞膜NMDA受体蛋白激光共聚焦显微镜亚细胞定位研究

Subcellular localization of NMDA receptor protein on neuron membrane by confocal laser scanning microscopy

目的 明确神经元膜NMDA受体蛋白分子在神经元表面的分布规律,对单个NMDA受体簇进行定位。方法 应用免疫荧光标记激光共聚焦显微镜方法,对原代培养大鼠皮层神经元进行观测。结果 与细胞核标记的碘化丙啶红色荧光对比,NMDA受体蛋白分子标记的FITC绿色荧光主要分布于神经元树突干膜和胞体近树突起始部分的胞膜上,呈明显的颗粒点状分布。结论 NMDA受体簇在单细胞上主要与突触后膜分布一致,位于树突起始部及树突干膜上,激光共聚焦技术可以精确地定位标记后单个膜蛋白分子。

Objective To identify the distribution of NMDA receptor protein on neuron membrane surface and the localization of single NMDA receptor cluster. Methods The primary culture of forebrain cortex neuron of rat was set up. Then the NR1 subunit protein molecule of NMDA receptor was combined with primary antibody and marked by FITC-IgG. Contrasted with red fluorescence marked by PI in nucleolus, the green fluorescence of NR1 membrane protein were inspired and observed by confocal laser scaning microscopy. Results NMDA receptor cluster combined with FITC molecule was observed obviously as some green dots congregating together and distinguished approximately. The green dots distributed mainly on the membrane surface of dendrite and origination section of dendritic stem. Conclusion NMDA receptor protein distributes on the membrane surface of dendrite and origination section of dendritic stem. It is consistent with the distributing rule of postsynaptic membrane. The confocal laser scanning microscopy could distinguish single NMDA receptor cluster marked by immunofluorescence.