实时荧光RT-PCR检测DLX2基因在小鼠前脑中的表达

Expression assay of DLX2 gene in mouse forebrain by real-time fluorescence RT-PCR

目的探讨DLX2基因在小鼠前脑神经干细胞的增殖、迁移及向多巴胺能神经元分化过程中的作用及意义。方法在小鼠胚胎16d(E16)、生后1d(P1),7d(P7)、21d(P21)4个时间点,分别取室管膜前下区(SVZa)、嘴侧迁移流(RMS)及嗅脑(O B)3个部位,应用SYBR Green I实时相对定量荧光逆转录聚合酶链反应(RT-PCR)技术检测DLX2基因在小鼠前脑中的表达情况,以DLX2基因相对表达值(Ratio)值大于1.4为差异表达。结果从部位看,OB区E16的表达较其他3个时间点低,RMS区P21的表达较其他3个时间点低,SVZa区E16及P1的表达较P7及P21高。从时间点看,在E16时间点3个部位的表达无差异,另3个时间点中OB较RMS及SVZa高表达。结论DLX2参与了小鼠前脑中神经干细胞的增殖、迁移及分化过程,其关系具有时间及部位特异性。

Objective To study the role of DLX2 gene in the proliferation, migration and dopaminergic differentiation of neural stem cells in mouse forebrain. Methods Expression of DLX2 in mouse forebrain was assayed at the 4 time points of embryonic day 16 (E16) to postnatal day 21 (P1, P7, P21), and the 3 sites including the anterior subventricular zone (SVZa), the rostral migratory stream (RMS),and the olfactory bulb (OB) by SYBR Green I real-time fluorescence RT-PCR, The relative expression value, ratio >1.4, of DLX2 was considered to be differential expression. Results DLX2 gene at E16 and P21 in OB and RMS respectively showed the lowest expression, while DLX2 gene at E16 and P1 in SVZa showed higher expression than that at P7 and P21. The DLX2 gene at E16 in the 3 regions showed equal expression, while at the other 3 time points, OB owned higher expression of DLX2 gene compared with that of RMS and SVZa. Conclusion These findings indicate that the DLX2 plays a time and region-specific role in the proliferation, migration and dopaminergic differentiation of neural stem cells in mouse forebrain.