激光共聚焦扫描显微镜和透射电镜观察大鼠纹状体内谷氨酸能突触连接的对比观察  

Comparative study on glutamatergic synaptic connections in rat striatum with laser scanning confocal microscopy and electron microscopy

作　　者：蔡青[1] 姬曼[1] 张进禄[1] 赵君朋[1] 

机构地区：[1]首都医科大学医学实验与测试中心,北京100069

出　　处：《中国组织化学与细胞化学杂志》2011年第3期236-240,共5页Chinese Journal of Histochemistry and Cytochemistry

基　　金：北京市自然基金资助项目资助(7062008)

摘　　要：目的探讨使用激光共聚焦扫描显微镜(Laser scanning confocal microscope,LSCM)观察大鼠纹状体内谷氨酸能突触连接的方法的可行性。方法 12只正常大鼠分为两组,6只大鼠进行纹状体中等棘刺神经元的CM-DiI单细胞标记,然后Ⅰ型囊泡膜谷氨酸转运体(vesicular glutamate transporter 1,VGluT1)免疫荧光标记,LSCM层扫后三维重建,观察VGluT1阳性位点在中等棘刺神经元树突上的分布。另外6只大鼠用TEM观察不对称性突触在纹状体神经元树突上的分布。对两种方法的结果进行比较。结果用LSCM和TEM方法观察到的纹状体神经元上谷氨酸能突触连接分布情况一致,没有统计学差异。但LSCM更具优越性的是,可以对图像进行三维重构,从而有利于对神经元之间突触连接的空间分布观察和定量分析。结论神经细胞荧光标记技术结合LSCM观察是考察纹状体神经元上谷氨酸能突触连接的有效方法。Objective To evaluate the advantage of the use of laser scanning confocal microscopy（LSCM） for observing the glutamatergic striatal synaptic connections in comparison with the observation with transmission electron microscopy（TEM）.Methods 12 normal adult rats were divided into two groups： 6 rats for LSCM or TEM each.With CM-DiI and VGluT1 immunofluorescence labeling,the distribution of the asymmetrical glutamatergic synaptic connections on the dendrites of striatal neurons was observed,and 3-D reconstruction was done.The 6 rats were studied with TEM.Results There is no significant difference in the distribution characters of the glutamatergic striatal synaptic connections obtained with either LSCM or TEM.However,the whole view of the synapses and dendrites and the three-dimensional reconstruction of synaptic connections between neurons can be obtained with LSCM but not TEM.Conclusion LSCM is an effective and quantitative technique to investigate the glutamate synaptic connections of striatal neurons.

关 键 词：激光共聚焦显微镜 透射电镜 免疫荧光染色 突触 

分 类 号：R322.811[医药卫生—人体解剖和组织胚胎学]