关键词: 核受体相关因子1, 神经干细胞, 多巴胺能神经元, 分化, 细胞培养, 大鼠

Abstract: ObjectiveTo study the effect of orphan nuclear receptor related factor 1 (Nurr1) gene on inducing neural stem cells (NSCs) derived from rat midbrain to differentiate into dopaminergic neuronsEM> in vitro/EM>. MethodsThe eukaryotic expression vector pEGFP-N1-Nurr1 was constructed by gene recombination technology and transfected into P3 NSCs through electroporation. The transfection efficiency was measured by fluorescence microscope, and the expression of Nurr1 in NSCs was detected by Western blotting. After differentiated for 3 weeks in vitro, the expression of tyrosine hydroxylase (TH) and microtubule associated proteins-2 (MAP-2) were detected by double-immunofluorescence labeling. ResultsThe transfection efficiency was about 35% 48 hours after transfection by electroporation, and Nurr1 protein was over-expressed 1 week after transfection with recombinant pEGFP-N1-Nurr1, increased about 5 times compared with the control group. Immunofluorescence showed that the proportion of TH positive neurons was 15.45%, which was markedly increased as compared with the control group. But there was no difference with the number of MAP-2 positive neurons. ConclusionThe findings demonstrate that NSCs can be induced to differentiate into TH positive dopaminergic neurons through Nurr1 gene overexpression.

Key words: Nuclear receptor related factor 1, Neural stem cell, Dopaminergic neuron, Differentiation, Cell culture, Rat