| Vitamin have a derivative which is Retinoic acid(RA),can induce various types of cells in a concentration-dependent manner in vitro,The way that active mi RNA can inhibit stem cell differential to neural cells.What is the mechanism of microRNAs function? In previous experiments,we found that miRNA-219 play a key role in RA-induced mouse J1 embryonic stem cell differentiating to neurons,and the targeted gene are Foxj3 and Zbtb18.In this study,through real-time quantitative PCR,western blot we induced with RA,Foxj3,Zbtb18 and downstream factors Olig1 and Olig2 can inhibit stem cells differential.For example,a high RA concentration increases the rate of neural differentiation,whereas a low RA concentration induces cardiomyocyte differentiation of mESCS.RA concentration regulates dorsoventral identity,that is,high RA concentration induces a dorsal phenotype,and low RA concentrations induces a ventral phenotype during in vitro mESC differentiation.On the basis of the results of the RNA-seq performed on Foxj3-ESCs and Zbtb18-ESCs,we constructed three gene co-expression networks and identified 17 core genes that are functionally related to neurodevelopment.16 of these genes were upregulated during neural differentiation,showing that most factors that dominate neural differentiation can be induced.We then demonstrated that Foxj3 and Zbtb18 are the molecular switches in the neurodevelopment system.we identified four critical gene,Olig1、Zic5、 Erbb2 and Olig2,by constructing gene co-expression networks with dif degree.And we found that Olig1 and Olig2 might be the most essential elements in neural differentiation of ESCs.Finally,we detected Olig1/2’s function in the control,over-express and Olig1/2-knockdown ESCs treated with RA or miR-219 mimics.This finding may be attributed to other factors or mechanisms involved in the miR-219-mediated neural differentiation apart from Olig1/2.Overall,we discovered that functionally important gene expression changes mediated by miRNA contribute to RA-induced differentiation.We demonstrated that Foxj3 and Zbtb18,two target genes of miR-219,regulate downstream gene Olig1、Zic5、 Erbb2 and Olig2 are the main controls in neural differentiation of ESCs.Our findings illustrate the mechanism of miRNA-mediated neural differentiation of ESCs,as well as the plasticity and dynamic nature of the gene regulatory networks during neural differentiation. |
