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.