冰冻切片染色法观察重组狂犬病病毒对乳鼠脑的感染和定位  

Virus detection and viral location in brain tissue of sulking mice infected recombinant rabies virus by frozen section

作　　者：黄莹[1] 龚锴[2] 唐青[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所脑炎室,北京100052 [2]清华大学生命科学院生物膜与膜生物工程国家重点实验室

出　　处：《中华实验和临床病毒学杂志》2010年第2期85-87,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家高技术研究发展计划863项目资助（2006AA02Z112）;2008年公益性行业科研专项经费预算执行情况表（200803014）;“十一五”重大专项（2009ZX10004-705）

摘　　要：目的冰冻切片染色法观察重组狂犬病病毒对乳鼠脑的感染和定位。方法将本实验室构建并拯救成功的人用狂犬病疫苗株重组病毒（CTN—GFP）脑内接种1日龄乳鼠和4周龄成年鼠，待感染乳鼠发病后至濒死期，通过心内灌流方法固定并制作脑组织冰冻切片，观察重组病毒在感染鼠的大脑海马组织中的绿色荧光蛋白的表达及分布。结果DAPI将所制作的冰冻切片中脑组织内细胞的胞核染成蓝色，在共聚焦显微镜下观察到受感染乳鼠的脑组织海马回位置有大量绿色荧光蛋白的表达。结论通过制作脑组织冰冻切片的方法可以清楚的观察到病毒的感染及其在海马回中的位置，同时为追踪病毒在体内的移行过程提供了一种新方法。Objective To observe the existence and location of the recombinant rabies virus in the hippocampus of the sulking mice infected recombinant rabies virus. Method A group of one-day-old sulking mice and 4-week-old mice were challenged with the CTN-GFP strain by intracerebral inoculation, frozen longitudinal transect sections of hippocampus were prepared from the suckling mice in order to observe the expression of the GFP protein and the location of the recombinant rabies virus. Results DAPI was performed to stain the cell nuclei in blue while GFP expression from CTN-GFP infected brain cells was observed under a confocal microscope. Conclusion The location of the rabies virus can be clearly observed by preparing frozen section of certain sites from the brain, and this method also provide a new tool to trace the route of spread of the rabies virus within the animal host.

关 键 词：冷冻 超薄切片术 狂犬病病毒 动物替代试验 

分 类 号：R512.99[医药卫生—内科学]