Study on expression of Hes1 mRNA during the differentiation of neural stem cells toward neurons
10.3969/j.issn.1671-8348.2013.33.029
- VernacularTitle:Hes1 mRNA 在神经干细胞向神经细胞分化过程中表达变化的研究
- Author:
Ruiying BAI
;
Na LU
- Publication Type:Journal Article
- Keywords:
hippocampus;
Hes1;
neuronal differentiation;
neural stem cells;
RT-PCR
- From:
Chongqing Medicine
2013;(33):4048-4050
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the expression of Hes1 mRNA during neural stem cells(NSC) differentiation toward neurons . Methods To establish the model of cultivation NSC in the hippocampal of newborn (24 h) SD rats ,and then to observe the mor-phology of NSC in the course of proliferation and differentiation .Before and after cellular induction ,the expression of Nestin and NSE were respectively measured to detect cell types by immunochemistry method .And flow cytometry was used to determine cell cycle phases ,so as to detect proliferative activity of these cells .Meanwhile ,the expression of Hes1 mRNA in NSC was determined by reverse transcription-PCR(RT-PCR) .Results The results demonstrated that NSC isolated from hippocampal showed vigorously clonal proliferation in vitro ,and positive Nestin expression .In addition ,the differentiated cells demonstrated positive NSE expres-sion .Flow cytometry analysis showed that the percentage of NSC in S phase was obviously higher than that of induced differentia-tion of all time(P<0 .01) ,which indicated that NSC were actively dividing induction before .Compared to NSC ,the percentages of cells in G0 G1 phases were increased significantly after neuronal differentiation (P<0 .01) ,which indicated that differented cells have arrested in G0 G1 phases .Meanwhile ,the results from RT-PCR showed that :Hes1 mRNA was expressed in NSC both before and af-ter induced differention .Compared to induction before ,the level of Hes1 mRNA expression in NSC during different stages of differ-entiation after induction were significantly decreased (P<0 .05) ,and Hes1 mRNA did not show any obvious changes among these stages of differentiation(P>0 .05) .Conclusion The high level of Hes1 mRNA was probably involved proliferation of NSC .How-ever ,low level of Hes1 mRNA might contribute to neuronal differentiation .