The expression of Mash1 and Hes1 during induce neural stem cells differentiating into dopaminergic neurons

Objective To build the in vitro modeld of NSCs differentiate into dopaminegic neurons,and further study the expression of Mash1 and Hes1 in this process.Methods Separated the telencephalon of rat embryos in sterile condition and cultured in vitro,take NSCs stained by Nestin immunofluorescence assay,divided into con=group and exp-group(GDNF+IL-1α) and induced into dopaminegic neurons,after 2 weeks stained by β-tubulinⅢ and TH,and counted the rate of positive cells of each group.q-PCR checked the expression of Mash1 and Hes1 in NSCs and during differentiated into dopaminegic neurons.Results IF showed that the rate of Nestin positive cells was(93.04±3.55)%,There were no statistical difference between the two comparison(P0.05).After differentiated 2 weeks,the rate of β-tubulinⅢ positive cells were:P4,con-group [(12.65±1.58)%],exp-group[(33.95±2.97)%],respectively,There were statistical difference between the two comparison(P0.05); the rate of TH positive cell were:P4,con-group [(1.46±0.41)% ],exp-group [(15.51±1.94)% ],There were statistical difference between the two comparison(P0.05).q-PCR result showed that after differentiation in P4 NSCs,Mash1 expression was higher than NSCs,there were statistical difference between each group,exp-group higher than con-group(P0.05),2 weeks higher than 1 week(P0.01); Hes1 expression was lower than NSCs,there were statistical difference compared with each group(P0.05).Conclusion The combined effect of GDNF and IL-1 was able to induce NSCs differentiating into dopaminergic neurons; Mash1 and Hes1 gene maybe involve in NSCs differentiated into dopaminergic neurons.