Roles of glycogen synthase kinase 3α and glycogen synthase kinase 3β in neural differentiation of mouse embryonic stem cell
Objective To investigate the roles of glycogen synthase kinase 3α(GSK3α)and glycogen synthase kinase 3β(GSK3β)in neural differentiation of embryonic stem cells(ESCs)in vitro.Methods Gsk3a or Gsk3b knockout ESCs lines were generated using CRISPR/Cas9 technology.These ESCs lines were then subjected to neural induction in vitro,and the differential gene expression of ESCs was analyzed using RNA sequencing(RNA-seq).Results Gsk3a-/-ESCs or Gsk3b-/-ESCs cells still maintained pluripotency and were capable of differentiating into neural stem cells(NSCs).However,Gsk3b-/-ESCs failed to form neural rosettes,and their NSCs were unable to further differentiate into neurons and glial cells.Analysis of mRNA expression of germ layer markers revealed slightly higher expression of neural ectoderm markers in Gsk3a-/-ESCs compared to the wildtype(WT)group,while the expression of endoderm markers in Gsk3b-/-ESCs was significantly higher than both the WT group and Gsk3a-/-ESCs(P＜0.001),with lower expression of neural ectoderm markers.RNA-seq results indicated changes in genes involved in the JAK-STAT,PI3K-Akt signaling pathway,as well as genes like Neurog1 and Hes1.Conclusions GSK3α inhibits neuronal differentiation while GSK3β promotes neuronal differentiation,and their mechanisms are associated with the JAK-STAT,PI3K-Akt signaling pathways,as well as Neurog1 and Hes1.
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