目的研究BMP2在SVZa神经干细胞向γ-氨基丁酸(GABA)能神经元分化中的调控作用。方法体外分离培养P0昆明小鼠室管膜下区(SVZa)神经干细胞,纯化传代培养3代后,使用不同浓度BMP2诱导SVZa神经于细胞,采用流式细胞仪检测不同浓度BMP2作用下SVZa神经干细胞分化为GABA能神经元的比例;另外利用活体荧光GFP标记GAD67特异性启动子,动态地研究BMP2在SVZa神经干细胞向GABA能神经元分化中的作用。在此基础之上,采用RT-PCR检测不同浓度BMP2作用下Mashl的表达。结果不同浓度BMP2作用组分化为GABA能神经元的比例均高于空白对照组,10ng/ml浓度的BMP2组比例最高;10ns/ml浓度BMP2组,GAD67-GFP标记阳性细胞数目明显高于对照组;10ng/ml浓度BMP2组Mashl表达高于其他组。结论BMP2促进SVZa神经干细胞向GABA能神经元的分化;10ng/ml浓度的BMP2显著促进Mashl的表达。
Objective To investigate the role of BMP2 in the differentiation of SVZa neural stem cells (NSCs) into GABAergic neurons. Methods The neural stem cells derived from anterior subventricular zone (SVZa) of Kunming mouse ( postnatal day 0, P0) were separated and cultured in vitro. The rate of GABAergic neurons in the differentiated cells of SVZa NSCs induced by BMP2 ( 1, 5, 10, 20, 100 ng/ml) were detected by flow cytometry (FCM). The role of BMP2 in the differentiation of SVZa NSCs into GABAergic neurons was studied by the GAD67 promoter labeled with GFP. The expression of Mashl in the SVZa NSCs induced by BMP2 of different concentration was detected by RT-PCR. Results FCM and the GAD67 promoter labeled with GFP confirmed that the rates of GABAergic neurons induced by BMP2 were higher than that of blank group. The expression of Mashl was enhanced by BMP2 in SVZa NSCs, especially by 10 ng/ml BMP2. Conclusion BMP2 promotes the differentiation of SVZa NSCs into GABAergic neurons.