| Objective:To detect the expression changes of Mash1 after the inhibition of Hes1 gene ,whenthe embryonic neural stem cells derived from rat spinal cord in fetal rats were differentiatedinto cholinergic neurons, to explore the differentiation mechanism of NSCs.Methods:The single cell suspensions, derived from rat embryos spinal cord at E17 d, wereplated into the serum-free medium. After the neural stem cells clone spheres formed, thespinal cord derived stem cells clone spheres were plated onto cover slips coated by polylysinein 6 well culture-plates. RA, SHH, NT-3 and BDNF as inducing factors were added into theculture medium in the induction group. In RNA interference group, the interference factorsiRNA was added into the culture medium, then,RA, SHH, NT-3 and BDNF as inducingfactors were added into the culture medium 6h later. The expression changes of Hes1, Mash1and ChAT gene were analyzed by real-time quantitative PCR during neural stem cellsdifferentiated into the cholinergic neurons after first week, second week and first week,second week after the Hes1 was interferd by siRNA.Results:The expression change of Hes1, Mash1 and ChAT mRNA were observed during theprogress. The expression of Hes1 and Mash1 mRNA have decreased in a significant mannerafter first week, and increased slightly after second week. And ChAT mRNA washigh-expressed after 1w and 2w in comparison to control group. After the Hes1 was interfedby siRNA, the expression of Hes1 was decreased. The expression of Mash1 have decreasedafter first week, and increased slightly after second week. ChAT mRNA was high-expressedafter 1w and 2w in comparison to the control group and the induction group.Conclusion: The Hes1 and Mash1 gene may play an important role in the process of NSCsbeing induced to differentiate into cholinergic neurons. Hes1 can regulate the differentiationprocess of NSCs, and was negative correlation with Mash1. The expression of Hes1 wasabated and the expression of Mash1 was increased, which were significant cause of NSCs inProliferation state steering cholinergic neurons. Objective:To detect the expression changes of Aldoc, Stmn1, Ywhag and Sept-9 after theinhibition of Hes1 gene in the process of the spinal cord stem cells in fetal rats specificallybeing differentiated into cholinergic neurons, to explore the roles of Aldoc, Stmn1, Ywhagand Sept-9 in the process of NSCs being differentiated into cholinergic neurons.Methods: the neural stem cells cultured in vitro were plated into 6 well cultivate plates coatedby poly-L-lysine, then siRNA was mixed into the serum-free medium to interfer the geneHes1. They were induced to differentiate into cholinergic neurons by adding RA, SHH, NT-3and BDNF to the culture medium. The expression changes of Aldoc, Stmn1, Ywhag andSept-9 gene were analyzed by real-time quantitative PCR during neural stem cellsdifferentiated into the cholinergic neurons after first week, second week and first week,second week after the Hes1 interferd by siRNA.Results: In the process of the spinal cord stem cells specifically being differentiated intocholinergic neurons, The expression of Aldoc mRNA have decreased in a significant mannerafter first week and second week. The expression of Stmn1 and Ywhag have decreased in asignificant manner after first week, The expression of them have increased slightly aftersecond week and was more than the control group. The expression of Sept-9 mRNA havedecreased after first week and second week, but the expression of Sept-9 mRNA after secondweek was more than that of first week. In the process of the spinal cord stem cells specificallybeing differentiated into cholinergic neurons after the inhibition of Hes1 gene, the expressionof Aldoc mRNA have decreased after first week and second week, but the expression of AldocmRNA was high-expressed in comparison to 1w after 2w; And the expression of them wasmore than the induction group. Stmn1 was high-expressed in comparison to control groupafter 1w and 2w, and was more than the induction group. In comparison to control group,Ywhag was low-expressed after 1w, and was high-expressed in comparison to 1w after 2w,but they were low-expressed in comparison to the induction group. Sept-9 was low-expressedin comparison to control group after 1w and 2w, and was low-expressed in comparison to theinduction group. Conclusion: The Aldoc, Stmn1, Ywhag and Sept-9 gene may play an role in the process ofNSCs being induced to differentiate into cholinergic neurons. Aldoc and Sept-9 may play anrole in maintaining the neural stem cells, Stmn1 and Ywhag may play an role in the latematuration process of cholinergic neurons; And Aldoc and Sept-9 were positive correlationwith Hes1, Stmn1 and Ywhag were negative correlation with Hes1. |
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