ADAM17异常表达对神经前体细胞迁移的影响  

The effect of ADAM17 abnormal expression on cell migration of neural precursors

作　　者：杨慈清[1,2] 李小英[1] 王聪睿 付苏雷[1] 李晗[1] 林俊堂[1,2] 

机构地区：[1]新乡医学院生命科学技术学院 [2]河南省医用组织再生重点实验室,新乡453003

出　　处：《神经解剖学杂志》2014年第6期683-687,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31000475);河南省教育厅科学技术研究重点项目(13A320866);新乡医学院重点研究领域招标课题资助项目(ZD2011-26)

摘　　要：目的：分析在胚胎发育过程ADAM17异常表达对神经前体细胞迁移的影响。方法：采用鸡胚活体电转和小鼠子宫内电转基因技术,在鸡胚发育的第5 d（E5）,将2μg/μl的p CAGGS-ADAM17和0.25μg/μl的p CAGGS-GFP质粒以1∶8混合液0.25-0.5μl准确地注射到视顶盖内,然后电转基因,E12时收集胚胎;小鼠胚胎发育的15 d（E15）进行子宫内电转,将2μg/μl的shRNA-ADAM17和0.25μg/μl的p CAGGS-GFP质粒以1∶8混合液0.25-0.5μl准确地注射到一侧脑室,电转基因后E20时收集胚胎。冰冻切片,采用免疫荧光组织化学和DAPI染色观察组织形态结构及相关蛋白的变化。结果：在鸡胚模型中,ADAM17的超表达并不会明显的引起神经前体细胞迁移的改变,对细胞核转录因子Pax7的表达也没有影响;在鼠胚模型中,ADAM17的抑制表达能够引起神经前体细胞迁移受阻,但并没有引起大脑皮层各层的结构的变化。结论：ADAM17的抑制表达对神经前体细胞的迁移具有抑制作用,但并不会引起大脑皮层结构的改变。Objective：To analyze the effect of ADAM17 abnormal expression on cell migration of neural precursors during embryonic development.Methods：Using in vivo electroporation,2 μg / μl p CAGGS-ADAM17 and 0.25 μg / μl p CAGGS-GFP plasmid were mixed by 1∶ 8,and 0.25 - 0.5 μl of which were accurately injected into the tectum of chicken embryos on embryonic day 5（ E5）.Electroporation was then conducted and the positive embryos were collected on E12.Similarly,2 μg / μl shRNA-ADAM17 and 0.25 μg / μl p CAGGS-GFP plasmid were mixed in 1∶ 8,and 0.25 - 0.5μl of the mixture was accurately injected into the lateral ventricle of the mouse embryos on E15.After electroporation,embryos were collected on E20.For frozen sections,immunofluorescence labeling and DAPI staining were carried out to observe the changes of the morphological structure and the related proteins expression.Results：Over-expression of ADAM17 could not cause a significant change of neural precursor＇s migration and of Pax7 expression in chicken embryo.Knockdown expression of ADAM17 could cause disorder of neural precursor＇s migration in the mouse embryo model,but induced no change in the structures of cerebral cortical layers.Conclusion：Knockdown expression of ADAM17 could inhibit the migration of neural precursor cells,but did not cause changes in the structure of the cerebral cortex.

关 键 词：ADAM17 神经前体细胞 鸡胚 小鼠胚胎 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]