敲除Pofut1不影响胚胎干细胞向神经元分化  

Silencing Pofut1 Gene Expression does not Affect Differentiation of Embryonic Stem Cells into Neurons

作　　者：来丽丽[1,2] 王玉华 姚雪莲[1,2] 王磊[2] 詹轶群[2] 葛常辉[1,2] 

机构地区：[1]安徽医科大学,合肥230032 [2]军事医学科学院放射与辐射医学研究所,北京100850 [3]美国爱因斯坦医学院内分泌系,纽约NY10461

出　　处：《中国生物化学与分子生物学报》2014年第6期591-596,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.30871416);北京市自然科学基金(No.5092021)资助~~

摘　　要：蛋白O-连接岩藻糖基转移酶1(Pofut1)基因缺失可导致Notch分子无法与配体结合并启动信号传递.为研究Pofut1基因对哺乳动物胚胎干细胞(ESC)向神经分化的影响,利用Pofut1基因敲除的胚胎干细胞与野生型胚胎干细胞,经体外培养诱导拟胚体(EB)分化为神经细胞,计数分化为神经细胞的比例,采用细胞免疫组化染色和real-time PCR等方法,分析神经细胞特异性标志分子的表达.结果显示,Pofut1基因缺失后,对EBC生长没有明显影响,分化过程中形成的拟胚体数量明显增多,分化的神经样细胞以及神经标志物分子的表达也明显多于对照组;Notch信号缺失对小鼠胚胎干细胞生长无明显影响,但可以促进ES细胞向神经细胞分化.Protein O-fucosyltransferase 1 （Pofut1）gene deletion disables Notch to combine its ligand for signal transmission. To study the gene function of Pofut1 in mammal embryonic stem cell （ESC） during differentiation, Pofut1 eliminated ESCs were evaluated by embryoid bodies （EB） counting when induced in vitro culture. Immunohistochemistry staining and real-time PCR were used to analyze the expression of nerve-specific molecules. The result showed that no significant difference between normal and Pofut1 eliminated ESCs. Meanwhile, the numbers of formed EB were significantly increased during the process of differentiation. The expression of neuronal and nerve cell markers were also elevated compared to the control group. No significant reduction of Notch signal upon the growth of mouse ESCs; and the ESCs were attenuated to differentiate into nerve cells.

关 键 词：NOTCH信号 蛋白O-连接岩藻糖转移酶1 胚胎干细胞 神经细胞分化 

分 类 号：Q28[生物学—细胞生物学]