鼠胚大脑皮层子宫内电穿孔法基因转移体系的建立  

The establishment of genetic transformation system in lateral cortex of mouse embryo via in utero electroporation

作　　者：俞华莉[1] 巨兴达[1] 王俊杰[1] 高倩倩[1] 伏秀青[1] 王兴智[1] 朱筱娟[1] 

机构地区：[1]东北师范大学遗传与细胞研究所,分子表观遗传学教育部重点实验室,长春130024

出　　处：《神经解剖学杂志》2009年第1期25-29,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(30670689);教育部新世纪优秀人才支持计划(NCET-07-0173);教育部博士点基金(20060200008);教育部回国人员启动基金资助项目

摘　　要：体内电穿孔法转基因技术作为一种新型有效的转基因方法,被广泛应用于发育生物学的研究,特别是中枢神经系统发育。本研究旨在通过子宫内电穿孔法,建立鼠胚大脑皮层体内基因转移体系:将带有GFP的表达载体pCAGGS-IRES-EGFP显微注入胚胎期14.5d的鼠胚侧脑室中,电极沿与脑中缝平行的方向夹持鼠脑,在电压30V,脉冲时间50ms,脉冲数5的条件下方波电击,将外源基因转移到室管膜层细胞。胚胎被重新放回母体内,继续发育3d。分离被转染的鼠胚脑组织,沿冠状面冰冻切片,在荧光显微镜下观察到外源蛋白GFP在鼠胚大脑皮层中的瞬时表达。鼠胚大脑皮层子宫内电穿孔法基因转移体系的建立将为进一步研究皮层发育、神经元迁移、轴突导向等以及神经发生过程中相关基因功能奠定基础。Recently, electroporation,as a new and effective gcne transfer method,has been applied to the study of mouse embryo developmental biology, especially to central nervous system development. In this study, in utero transfection plasmid DNA method was set up to embryonic mouse. The GFP expression vector pCAGGS-IRES-EGFP was microinjected into the lateral cerebral ventricle of E14.5 mouse embryos through the uterine wall. Then, the 30 V square-wave pulse was delivered across the head for 5 times with interval 50 ms by electrodes in parallel with the midian rapbes. Embryos were then allowed to develop for three days. The slices of transfected brain were checked under a fluorescence dissection microscope. The results showed that GFP was expressed in the lateral cerebral cortex. This system is of great significance to the further studies of cerebral cortex development, neuronal migration, axonal guidance and gain-of-function and loss-of-function induction.

关 键 词：子宫内电穿孔法 显微注射 大脑皮层 鼠胚 

分 类 号：Q789[生物学—分子生物学]