WNT/β-catenin信号调节神经干细胞向少突胶质-GABA能神经元分化的体外研究  

Regulation of oligodendrocyto-GABAergic neuron lineage differentiation from neural stem cells by Wnt/beta-catenin signaling in vitro

作　　者：田士来 赵宝成[2] 王甲义 宁伟东[1] 汪仁聪 陈伟[1] 王亚周[3] 屈延[1] 

机构地区：[1]第四军医大学西京医院神经外科,西安710032 [2]解放军总政治部门诊部,北京100034 [3]第四军医大学基础部神经生物学教研室,西安710032

出　　处：《神经解剖学杂志》2015年第2期165-170,共6页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31271583);国家优秀青年科学基金(81222015);教育部新世纪优秀人才支持计划(NCET-12-1004)

摘　　要：目的:在体外研究Wnt/β-catenin信号对于少突胶质-GABA能神经元前体细胞分化的影响。方法:培养胎鼠中脑神经干细胞,在Wnt3a刺激或过表达β-catenin的条件下进行诱导分化,采用免疫细胞化学染色和Western Blot检测Wnt/β-catenin信号激活时,少突胶质-GABA能神经元共同前体细胞标记蛋白DLX2,少突胶质细胞标记蛋白NG2,GABA能神经元标记蛋白GAD67的表达。结果:Western Blot显示Wnt-3a处理上调β-catenin和Wnt/β-catenin信号下游靶基因cyclin D1和Axin2的表达;Wnt3a处理和过表达β-catenin的神经干细胞可显著上调Dlx2,NG2和GAD67阳性细胞的百分率(P<0.05)。Wnt3a刺激和β-catenin过表达均抑制神经干细胞向星型胶质细胞方向的分化。结论:Wnt/β-catenin信号可在体外促进神经干细胞向GABA能神经元和少突胶质细胞方向分化。Objective： To explore the effects of Wnt/β-catenin signaling on the differentiation of oligodendrocyteGABAergic neuron lineage from neural stem cells in vitro. Methods： Neural stem cells were primarily cultured from embryonic midbrain. Under the conditions of Wnt3 a stimulation and β-catenin over-expression,neural stem cells were induced to differentiate. Immunocytochemistry and Western Blot for Dlx2（ a marker for the common progenitor of oligodendrocyte and GABAergic neurons）,NG2（ a marker of oligodendrocyte precusors） and GAD67（ a marker of GABAergic neurons） were performed to examine the differentiation of oligodendrocyte and GABAergic neurons. Results： Western Blot showed that Wnt3 a significantly increased the expression of β-catenin,and Cyclin D1 and Axin2,two Wnt / β-catenin signaling downstream genes. Both Wnt3 a stimulation and beta-catenin over expression significantly increased the percentages of Dlx2,NG2 and GAD67-positive cells（ P 〈 0. 05）,and showed significantly inhibitory effects on the differentiationof neural stem cells into astrocyte. Conclusion： Wnt / β-catenin signaling promotes the generation of oligodendrocyte and GABAergic neurons from neural stem cells in vitro.

关 键 词：神经干细胞 Wnt/β-catenin信号 GABA能神经元 少突胶质细胞 

分 类 号：R741[医药卫生—神经病学与精神病学]