神经细胞膜NMDA受体蛋白单分子胶体金标记扫描电镜定位研究  

A study of localization of NMDA receptor protein labelling with colloid-gold on membrane by Scanning Electron Microscopy

作　　者：郁毅刚[1] 徐如祥[1] 姜晓丹[1] 柯以铨[1] 

机构地区：[1]第一军医大学珠江医院,广州510282

出　　处：《解放军医学杂志》2004年第8期722-724,共3页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金 (编号 30 1 70 961 );军队"十五"重点科研基金 (编号0 1Z0 54)资助课题

摘　　要：目的　探讨神经元膜NMDA受体蛋白分子在神经元膜表面的分布规律 ,对单个膜NMDA受体蛋白进行定位。方法　应用免疫胶体金技术、场发射枪扫描电镜方法 ,对原代培养大鼠皮层神经元膜表面NMDA受体进行定位标记、观测。结果　NMDA受体蛋白分子标记的胶体金颗粒主要分布于神经元胞体两端近树突起始部分的胞膜脂筏小窝内 ,呈明显的颗粒点状分布。结论　NMDA受体蛋白在细胞膜上主要与突触后膜分布一致 ,位于树突起始部及树突干膜脂筏微区内 ,免疫胶体金标记、场发射枪扫描电镜技术可以精确地定位标记后单个NMDA受体膜蛋白分子。Objective To investigate the distribution rule of NMDA receptor protein localizated on neuron membrane surface. Methods After the primary culture model of forebrain cortex neuron of rat had been set up,the NR1 subunit protein molecule of NMDA receptor has been combined with primary antibody IgG,and marked by Staphylococcus protein A-colloid gold particles (SPA-G). Then the neuron was observed with Field-emission SEM,and the content of AU element was calcalated by software. Results The cluster of NMDA receptor combined with SPA-G particulates was clearly observed as some globular being distributed mainly under some niches of lipid rafts,mainly localized at originating section of dendritic protuberance of neuron membrane surface. Conclusion NMDA receptor molecule protein was distributed on the membrane surface lipid rafts of originating section of dendritic protuberance of neuron. It was in accord with the distribution rule of postsynaptic membrane on neuron. The Field-emission SEM could distinguish single NMDA receptor melocule cluster after being marked with SPA-G molecule in neuron.

关 键 词：神经细胞 受体 NMDA 免疫胶体金技术 场发射枪扫描电镜 脂筏 

分 类 号：Q73[生物学—分子生物学]