应用病毒示踪技术观察大鼠两侧背根神经节的纤维联系  

Observation the fiber connections of bilateral dorsal root ganglia in rats with virus tracing technique

作　　者：刘法东 段红梅[2] 郝鹏[2] 赵文 高钰丹 杨朝阳[2,3] 李晓光 Liu Fadong;Duan Hongmei;Hao Peng;Zhao Wen;Gao Yudan;Yang Zhaoyang;Li Xiaoguang(Beijing Key Laboratory for Biomaterials and Neural Regeneration,School of Biological Science and Medical Engineering,Beihang University,Beijing 100083;Department of Neurobiology,Capital Medical University,Beijing 100069;Beijing International Cooperation Bases for Science and Technology on Biomaterials and Neural Regeneration,Beijing Advanced Innovation Center for Biomedical Engineering,Beihang University,Beijing 100083,China)

机构地区：[1]北京航空航天大学,生物与医学工程学院,生物材料与神经再生北京市重点实验室,北京100083 [2]首都医科大学神经生物学系,北京100069 [3]北京航空航天大学,生物医学工程高精尖创新中心,生物材料与神经再生北京市国际科技合作基地,北京100083

出　　处：《神经解剖学杂志》2020年第3期233-239,共7页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31730030,31971279,31670988,31771053,31650001,31320103903);国家重点研发计划(2017YFC1104001;2017Y FC1104002);北京市科技计划(Z181100001818007);北京市教育委员会2018年度科技计划重点项目(KZ201810025030);中国博士后科学基金面上项目(230210465)。

摘　　要：目的:通过伪狂犬病毒(PRV)逆行示踪技术观察Wistar大鼠两侧背根神经节(DRG)之间神经元的联系,寻找两侧DRG神经元交互支配的形态学基础.方法:选用18只正常成年雌性Wistar大鼠,分为单侧PRV组、双侧PRV组、霍乱毒素B亚单位(CTB)组(n=6).单侧PRV组将2μl滴度为1×10^10的PRV-EGFP注射到左侧坐骨神经;双侧PRV组将2μl滴度为1×10^10的PRV-EGFP注射到左侧坐骨神经上,同时2μl滴度为1×10^10的PRV-mRuby注射到右侧坐骨神经上;CTB组将2μl浓度为1μg/μl的CTB注射到左侧坐骨神经上.5d后观察病毒标记结果.结果:单侧PRV组在左侧DRG可见大量被PRV-EGFP标记的神经元,右侧DRG也观察到大量被PRV-EGFP标记的神经元,但数量明显少于左侧(P<0.01).双侧PRV组在左侧DRG可见大量被PRV-EGFP和PRV-mRuby标记的神经元,且有部分神经元被PRV-EGFP和PRV-mRuby同时标记.CTB组大鼠L4脊髓前角神经元被CTB标记,DRG中枢突大量纤维终末被CTB标记.此外,L2脊髓中背根神经节中枢突的大量纤维终末被CTB标记.单侧PRV组大鼠L4脊髓前角神经元被PRV-EGFP标记,腰骶髓后连合核(DCN)有大量神经元被PRV-EGFP标记,L2腰骶髓后连合核也有大量神经元被PRV-EGFP标记.结论:Wistar大鼠的左右侧DRG神经元之间存在交互支配的现象,两侧DRG神经元之间并没有直接的突触联系,而是通过后连合核等中间神经元跨突触联系.Objective:The retrograde tracing technique of pseudorabies virus(PRV)was used to observe the neuron connection between the dorsal root ganglia(DRG)on both sides of Wistar rats,in order to find the morphological basis of the interactive domination of DRG neurons on both sides.Methods:Eighteen normal female Wistar rats were selected and divided into unilateral PRV group,bilateral PRV group,and cholera toxin B subunit(CTB)group(n=6).The unilateral PRV group injected 2μl of PRV-EGFP with a titer of 1 ×10^10 into the left sciatic nerve;the bilateral PRV group injected 2μl of PRV-EGFP with a titer of 1 ×10^10 into the left sciatic nerve,while the 2μl titer was 1 ×10^10 PRV-mRuby was injected into the right sciatic nerve;the CTB group injected 2μl of CTB at a concentration of 1μg/μl into the left sciatic nerve.The results of virus labeling were observed after 5 days.Results:In the unilateral PRV group,a large number of neurons labeled with PRV-EGFP were observed on the left DRG,and a large number of neurons labeled with PRV-EGFP were also observed on the right DRG,but the number was significantly less than that on the left(P<0.01).In the bilateral PRV group,a large number of neurons labeled with PRV-EGFP and PRV-mRuby were found in the left DRG,and some neurons were simultaneously labeled with PRV-EGFP and PRV-mRuby.The anterior horn neurons of the L4 spinal cord in the CTB group were labeled with CTB,and a large number of fibers in the DRG central process were labeled with CTB.In addition,a large number of fibrous terminals in the central process of the dorsal root ganglion in the L2 spinal cord were labeled with CTB.In the unilateral PRV group,anterior horn neurons of L4 spinal cord were labeled with PRV-EGFP,and a large number of neurons were labeled with PRV-EGFP in the dorsal commissural nucleus(DCN).Meanwhile,a large number of neurons in the dorsal commissural nucleus of the L2 spinal cord were also labeled with PRV-EGFP.Conclusion:There was a phenomenon of cross-domination between the left and right DRG

关 键 词：伪狂犬病毒 逆行示踪技术 背根神经节 突触联系 大鼠 

分 类 号：Q42[生物学—神经生物学]