背根神经节电穿孔转染神经元技术的改进  

Improving neuronal transfection by electroporation of dorsal root ganglion

作　　者：齐士斌 马艳霞[2] 张浩楠 马进进 徐金辉 赛吉拉夫 Qi Shibin;Ma Yanxia;Zhang Haonan;Ma Jinjin;Xu Jinhui;Saijilafu(Department of Orthopedics,the First Affiliated Hospital,Soochow University,Suzhou 215006,Jiangsu Province,China;Orthopedics Institute of Soochow University,Suzhou 215007,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院骨科,江苏省苏州市215006 [2]苏州大学骨科研究所,江苏省苏州市215007

出　　处：《中国组织工程研究》2020年第19期3042-3047,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81772353,81571189)，项目负责人：赛吉拉夫~~

摘　　要：背景:背根神经节电穿孔技术是一种研究神经再生的高效基因转染方法,以往背根神经节电穿孔的电压条件使得标记的神经元及轴突数目较少,统计误差较高。目的:提高神经元及其轴突的标记率,为周围神经再生的研究提供理论依据。方法:以增强型绿色荧光蛋白为观察指标优化背根神经节电穿孔技术在神经元轴突再生方面的应用。将ICR小鼠随机分为2组,分别行背根神经节电穿孔手术,检测在35 V和60 V电压干预下神经元及其轴突的标记率。结果与结论:①35 V和60 V电压未造成神经元明显死亡;②与35 V电压相比,60 V电压条件下标记的神经元及其轴突数量显著增加,且通过损伤部位的轴突数量明显增多(P<0.05);③60 V电压对实验动物未造成行为功能损害;④结果表明,60 V电压能够使神经元及其轴突的标记率增高,其结果能够为周围神经元轴突再生研究提供依据。BACKGROUND:Electroporation of dorsal root ganglion is a high-efficiency gene transfection method to study nerve regeneration.In the past,the voltage condition of dorsal root ganglion electroporation resulted in a reduction in the number of labeled neurons and axons,with a high statistical error.OBJECTIVE:To improve the marker rate of neurons and their axons,and to provide theoretical basis for the study of peripheral nerve regeneration.METHODS:The enhanced green fluorescent protein was as an outcome measure to optimize dorsal root ganglion electroporation in axonal regeneration.ICR mice were randomly divided into two groups,respectively,and underwent dorsal root ganglion electroperforation surgery to detect the labeling rates of neurons and their axons under the intervention of 35 and 60 V voltages.RESULTS AND CONCLUSION:Voltages at 35 and 60 V did not cause significant neuronal death.Compared with 35 V voltage,60 V voltage significantly increased the labeling rate of neurons and their axons as well as the number of axons passing through the injured site(P<0.05).The 60 V voltage did not damage the behavioral function of the experimental animals.These results suggest that 60 V voltage can increase the labeling rate of neurons and their axons,providing a basis for the study of axonal regeneration in peripheral nerves.

关 键 词：背根神经节 轴突 电穿孔 电压 周围神经 标记率 

分 类 号：R459.9[医药卫生—治疗学] R741.05[医药卫生—临床医学]