活体小鼠坐骨神经的形态学观察  

In vivo observation of mouse sciatic nerve morphology

作　　者：王天兵[1] 张丞[1] 寇玉辉[1] 王瑾[1] 王静[1] 王延华[2] 姜保国[1] 

机构地区：[1]北京大学人民医院创伤骨科 100044 [2]云南昆明医学院

出　　处：《中华手外科杂志》2009年第5期301-303,共3页Chinese Journal of Hand Surgery

基　　金：国家重点基础研究发展计划-973计划基金资助项目（2003CB515306）;国家自然科学基金杰出青年基金资助项目（30625036）

摘　　要：目的使用表达绿色荧光蛋白的转基因小鼠，建立用激光共聚焦显微镜活体、实时观察周围神经形态的方法。方法将转基因小鼠麻醉后，分离显露坐骨神经，并将小鼠固定于激光共聚焦显微镜操作台，使显露的坐骨神经与载玻片贴合，从而观察活体小鼠坐骨神经的形态学特点。测量神经干、神经束及神经纤维的直径，并计算神经束及神经纤维的数量；将小鼠坐骨神经切断或钳夹，造成神经损伤模型，在损伤即刻及损伤后2h，观察损伤神经近端、损伤处及损伤远端形态的改变。结果正常小鼠坐骨神经神经干直径为800．0μm，神经束直径为70．0-100．0舯，神经束数量为2个，神经纤维直径为7．5μm，神经纤维数量为70～200个。神经损伤即刻及损伤后2h，损伤近端处可见神经纤维逐渐出现空泡样变性，神经纤维走向逐渐紊乱，部分纤维中断。损伤段神经纤维完全扭曲，空泡样变性明显，并出现神经纤维崩解，神经连续性完全中断。损伤远端250．0μm及500．0μm处，神经纤维荧光强度逐渐增强，结构恢复，排列逐渐整齐。结论绿色荧光蛋白转基因小鼠结合激光共聚焦显微镜在体成像技术为实现活体实时观测周围神经提供了良好的技术平台，可用于观察小鼠正常及损伤后周围神经结构的形态和变化。Objective To observe the uhrastructure of sciatic nerve of GFP transgenie mice under normal and damaged conditions as well as to explore the applications of laser scanning eonfocal microscope for in vivo and real time observation of peripheral nerve morphology. Methods The GFP mice were anesthetized and the sciatic nerve was exposed. The mouse was then mounted on the platform of a laser scanning eonfoeal microscope in a way that the nerve was in direct contact with the slide for real time morphological observation in vivo. The diameters of the nerve trunk, nerve fascicles and nerve fibers were measured and the number and morphology of nerve fascicles and nerve fibers were recorded. Then the sciatic nerve was subjected to crash or transection to create a nerve injury model. Morphological changes of the nerve proximal to, at and distal to the injury site were observed immediately after the injury and 2 hours later by laser scanning confocal microscope in vivo. Results The diameter of normal sciatic nerve trunk of mice was about 800μm, the diameter of nerve fascicle and nerve fiber were 70 to 100 tan and 7.5 μm respectively. Each normal sciatic nerve trunk contained about 2 fascicles and 70 to 200 nerve fibers. Immediately after and 2 hours after nerve injury vacuolization was seen at the nerve proximal to the injury site. The nerve fibers were disoriented and disconnection of the fibers was also seen. Nerve fibers of the nerve at the injury site were distorted, with vaeuolization and breaking down of the fibers. Nerve continuity was lost. At the nerve 250μm and 500 μm distal to the injury site, the nerve fiber fluorescence intensity gradually increased and the nerve fiber structure restored gradually. Orientation of the fibers tended to be organized. Conclusion The use of GFP transgenic mice and laser scanning confocal microscope provides a technical platform for in vivo real time observation of peripheral nerves. It can be applied to observe the morphological features and changes of mouse peripheral nerves.

关 键 词：小鼠 活体解剖 坐骨神经 绿色荧光蛋白 

分 类 号：R686[医药卫生—骨科学]