离体电转染pCAGGS-EGFP对小鼠视网膜神经节细胞的标记  

Electrotransfection using pCAGGS-EGFP plasmid for labeling retina neuroganglion in mice

作　　者：施圆圆[1] 何士刚[2] 李书华[3] 

机构地区：[1]广州医科大学生物技术实验中心,广东广州510182 [2]中科院生物物理研究所,北京100101 [3]广州医科大学病理教研室,广东广州510182

出　　处：《广州医学院学报》2013年第4期10-12,共3页Academic Journal of Guangzhou Medical College

基　　金：广州市教育局科技攻关项目:08A096

摘　　要：目的：通过离体电转染将GFP对胚胎期小鼠视网膜神经节细胞（RGC）进行标记来观察其形态．方法：剥离胚胎小鼠E14的视网膜连同晶状体，放入定制的电转杯中进行电转染，以8～16V电压将pCAGGS—EGFP质粒导入RGC，体外培养视网膜1～5d，观察绿色荧光蛋白GFP的表达情况。结果：以12V，50ms，950InS的间隔，5个脉冲转染效率最高，为81．25％：20～144h期间GFP依次由胞体向轴突、树突扩散分布。结论：在体外培养基中培养1～5d．pCAGGS—EGFP经过最适电压转染后能够在小鼠视网膜神经节细胞中持续扩散表达.Objective ：To label retina neuroganglion cells （ RGC ） with green fluorescent protein （GFP） in embryonic mice via electrotransfection, which entailed subsequent morphological assessments. Methods： The retina and lens in El4 mice were extracted in a specially prepared container for subsequent electrotransfection. The pCAGGS-EGFP plasmid was introduced into RGC by using a potential of 8-16 volts. This was followed by in vitro incubation of the retina for 1 to 5 days to determine the expression of GFP. Results ： Electrotransfection with a potential of 12 volts and at 50ms and 950ms intervals for 5 pulsations was characterized by the highest efficiency （81.25 % ）. GFP was distributed to the axons and dendrites between hour 20 and 144. Conelusion： pCAGGS- EGFP has a steady and effective expression in the mouse retinal ganglion cell （ RGC ） at embryonic stage for 5 days by electroporation in vitro at an optimal voltage.

关 键 词：视网膜神经节细胞 离体电转染 pCAGGS—EGFP质粒 体外培养 

分 类 号：R774.1[医药卫生—眼科]