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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 abnormal expression of ADAM17 on neural precursor cell migration during embryonic development. Methods: In the fifth day (E5) of chick embryo development, 2 μg / μl of pCAGGS-ADAM17 and 0.25 μg / μl of pCAGGS-GFP plasmid were digested with 1 : 8 mixture of 0.25 ~ 0.5μl accurately injected into the optic disc cap, and then electroporated gene, E12 embryos collected; embryonic development 15d (E15) intrauterine electroporation, 2μg / μl shRNA-ADAM17 and 0.25 μg / μl of pCAGGS-GFP plasmid was accurately injected into a lateral ventricle at 0.25-0.5 μl of a 1: 8 mixture and the embryos were harvested electroporated at E20. Frozen sections were obtained. The changes of histomorphology and related proteins were observed by immunofluorescence histochemistry and DAPI staining. RESULTS: In chick embryo model, overexpression of ADAM17 did not significantly change the migration of neural precursor cells and had no effect on the expression of nuclear transcription factor Pax7. In mouse embryo model, the inhibitory expression of ADAM17 could induce neuronal Precursor cell migration blocked, but did not cause changes in the structure of the various layers of the cerebral cortex. Conclusion: The inhibitory expression of ADAM17 has an inhibitory effect on the migration of neural progenitor cells, but does not change the structure of cerebral cortex.