利用子宫内胚胎电转技术研究Frizzled 10在大脑皮质发育中的功能  

Preliminary study of the function of Frizzled 10 in the development of cerebral cortex by in utero electroporation

作　　者：刘斌[1] 顾小春[1] 

机构地区：[1]东南大学生命科学研究院发育与疾病相关基因教育部重点实验室,江苏南京210009

出　　处：《东南大学学报（医学版）》2011年第3期401-406,共6页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(81000485);东南大学新进博士基金资助项目(9231000101)

摘　　要：子宫内胚胎电转技术目前被广泛应用于神经发育的相关研究。本研究利用子宫内胚胎电转的方法对Frizzled 10基因在小鼠大脑皮层发育过程中的作用作了初步的探讨。首先利用PCR扩增获得Frizzled 10的全长cDNA,将cDNA连接到电击载体pCAGIG,构建异位表达载体pCAGIG-Frizzled 10。建立子宫内电击技术:在胚鼠E 14.5 d时,将异位表达载体pCAGIG-Frizzled 10在电压35 V、脉冲时间50 ms、脉冲数5的条件下将质粒电击到背侧皮质的脑室区,4 d后收取胚鼠脑组织,进行冠状切片后在荧光显微镜下检测外源蛋白GFP在鼠胚背侧皮质的表达情况。结果显示,与对照相比,Frizzled 10的异位表达参与了皮质发育的调控。该研究为我们进一步研究Frizzled 10的功能提供了新的线索。In utero electroporation has been applied to studies of the cortical development.To elucidate the role of Frizzled 10 in the cortical development,Frizzled 10 was ectopicly expressed by means of in utero electroporation.In order to get the ectopic expression vector pCAGIG-Frizzled 10,we first amplified Frizzled 10 full length cDNA by PCR,and then inserted the cDNA fragment into the pCAGIG vector.pCAGIG-Frizzled 10 DNA was then microinjected into the lateral cerebral ventrical of the dorsal cortex at the developmental stage of E 14.5,35 V pulse was delivered for 5 times with interval 50 ms by regulate the direction of the electrodes to make the plasmid DNA get into the lateral ventricular zone.Mouse embryoes were allowed to develop for four days.The coronal slices of the electroporation brain were examined by fluorescene microscope.The reporter gene GFP was expressed in the dorsal cortex.Our results show the neuron migration was affected after Frizzled 10 ectopic expression,indicating Frizzled 10 is important for the cortical development.

关 键 词：FRIZZLED 10 大脑皮质 子宫内胚胎电转 

分 类 号：R-332[医药卫生] Q786[生物学—分子生物学]