糖原合成激酶3α和糖原合成激酶3β在小鼠胚胎干细胞神经分化中的作用  

Roles of glycogen synthase kinase 3α and glycogen synthase kinase 3β in neural differentiation of mouseembryonic stem cell

作　　者：邓卫岭 张旭辉[1] 陈子睿 黄伊华 胡亚芳[1] 吴永明[1] Deng Weiling;Zhang Xuhui;Chen Zirui;Huang Yihua;Hu Yafang;Wu Yongming(Nanfang Hospital,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学南方医院,广东广州510515

出　　处：《中国临床解剖学杂志》2024年第6期660-665,共6页Chinese Journal of Clinical Anatomy

基　　金：国家自然科学基金(8207053359);广东省自然科学基金(2019A1515011760)。

摘　　要：目的 探讨GSK3α和GSK3β在胚胎干细胞体外神经分化中的作用。方法 利用CRISPR/Cas9技术构建Gsk3a或Gsk3b敲除的胚胎干细胞,在体外对其进行神经诱导分化,并通过转录组测序(RNA-seq)分析胚胎干细胞的基因表达差异。结果 Gsk3a^(-/-)或Gsk3b^(-/-)胚胎干细胞仍然具有多能性,能够分化成神经干细胞。然而,Gsk3b^(-/-)无法形成神经花环,其神经干细胞也不能进一步分化成神经元和胶质细胞。检测三胚层标志物的mRNA表达发现,Gsk3a^(-/-)神经外胚层标志物的表达略高于野生(WT)组,而Gsk3b^(-/-)内胚层标志物的表达远高于WT组和Gsk3a^(-/-)(P<0.001),神经外胚层标志物的表达较低。RNA-seq结果显示,涉及JAK-STAT、PI3K-Akt信号通路以及Neurog1、Hes1等基因的变化。结论 GSK3α抑制神经分化而GSK3β促进神经分化,其机制与JAK-STAT、PI3K-Akt信号通路以及Neurog1、Hes1有关。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 GSK3αinhibits neuronal differentiation while GSK3βpromotes neuronal differentiation,and their mechanisms are associated with the JAK-STAT,PI3K-Akt signaling pathways,as well as Neurog1 and Hes1.

关 键 词：糖原合成激酶3α 糖原合成激酶3β 胚胎干细胞 神经分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]