大鼠脑室管膜下区包埋前免疫电镜双标技术的探索  

Exploration of pre-embedding double labeling technique under immune electron microscopy in subventricular zone of rats

作　　者：于红丽[1] 李志宁[2] 董富兴[1] 张伟[2] 董红燕[1] 

机构地区：[1]徐州医学院神经生物学研究中心,江苏省221002 [2]徐州医学院第二附属医院神经内科

出　　处：《江苏医药》2013年第14期1627-1629,共3页Jiangsu Medical Journal

基　　金：江苏省高校自然科学基础研究计划项目(07KJB310119);徐州医学院科研课题(2010KJ18)

摘　　要：目的观察脑室管膜下区(SVZ)神经干细胞上N-甲基-D-天冬氨酸受体亚单位1(NR1)的超微分布。方法成年大鼠灌注固定,冠状面振动切片,片厚150μm。取含SVZ区域的脑片,行巢蛋白(nestin)和NR1双标免疫组织化学染色。其中,巢蛋白为辣根过氧化物酶标记,NR1为免疫金-银法标记。取1.5mm×1.0mm大小的SVZ双标阳性组织,常规电镜制样、观察、拍照。结果电镜下观察到标记NR1的纳米金-银标颗粒电子密度高,呈颗粒状,分布于细胞膜上,偶见于胞质中;巢蛋白免疫阳性产物呈絮状,分布于胞质中。结论包埋前免疫电镜双重标记技术对组织的抗原性保存较好,免疫金颗粒对抗原定位较精确,有利于研究两种递质在同一个细胞或终末内的共存或分析递质与其相应受体之间的联系,特别是对于中枢神经系统的研究有独到之处。Objective To observe the distribution of N-methyl-D-aspartate receptor 1（NR1） in the neural stem cells of subventricular zone（SVZ）.Methods Three adult rats were perfused and fixed,and the cerebral slices with the thickness of 150 μm were obtained by vibratome.The slices containing SVZ were taken and stained by nestin（horseradish peroxidase labelled） and NR1（immunogold-silver labelled） double immunohistochemistry.Then the positive tissues of SVZ with the size of 1.5 mm 1.0 mm were observed and photographed under electron microscopy.Results Under immune electron microscopy,high electron density of immunogold-silver labelled NR1 presented particle was distributed mainly on the cell membrane and rarely in the cytoplasm,while positive product of nestin presented floccus located in the cytoplasm.Conclusion Pre-embedding double labeling technique under immune electron microscopy and immunogold particle have greater preservation and localization of the antigen in many tissues,respectively,which is helpful for studying the co-existence of two transmitters in single cell or axon terminal and analyzing the relationship between a transmitter and its relevant receptor,especially in the central nervous system.

关 键 词：纳米金-银加强法 免疫电镜 神经干细胞 N-甲基-D-天冬氨酸受体 

分 类 号：R331[医药卫生—人体生理学]