Brn-4抗体对大鼠海马神经干细胞分化为神经元的影响

为探讨Brn-4在海马神经干细胞(NSCs)向神经元分化过程中的作用,分别制备成年SD大鼠海马伞切割侧和正常侧海马提取液;将从鼠胚海马中分离、克隆和扩增的神经干细胞球分成6组:(1)切割组:加入含切割侧海马提取液的DMEM/F12培养基;(2)切割阻断组:在切割组培养基中加入Brn-4多克隆抗体;(3)正常组:加入含正常侧海马提取液的DMEM/F12培养基;(4)正常阻断组:在正常组培养基中加入Brn-4多克隆抗体;(5)空白组:加入单纯的DMEM/F12培养基;(6)空白阻断组:在空白组培养基中加入Brn-4多克隆抗体。培养后第14d对NSCs分化的神经元进行MAP-2免疫荧光检测,图像处理系统计数各组MAP-2阳性神经元数、处理胞体面积和细胞周长,STATA7.0软件进行统计分析。结果显示:各组以上三项形态学指标从优到劣的排列顺序为:切割组、正常组、切割阻断组、正常阻断组、空白组和空白阻断组。统计分析结果表明,除空白组与空白阻断组三项指标无显著性差异外,其余各组间差异均有显著性意义(P<0.05)。上述结果提示,Brn-4多克隆抗体在体外能部分地阻断切割海马伞侧海马提取液促进NSCs向神经元分化的作用,因而Brn-4在海马NSCs向神经元分化过程中可能发挥重要作用。

Effects of Brn-4 antibody on the differentiation of neural stem cells into neurons in the rat hippocampus

To explore the role of Brn-4 in the process of the neural stem cells (NSCs) differentiating into neurons, the extracts from fimbria transected and intact hippocampus of adult SD rats were respectively obtained. NSCs isolated and expanded from the hippocampus of embryonic rat were divided into 6 groups as follows: (1) the transection group: DMEM/F12 medium containing extracts of the fimbria-transected hippocampus was added; (2) the transection-interdiction group: the Brn-4 polyclonal antibody was added on the basis of transection group; (3)the normal group: DMEM/F12 medium containing extracts of the normal hippocampi was used; (4)the normal-interdiction group: the Brn-4 polyclonal antibody was added on the basis of the normal group; (5)the control group: DMEM/F12 medium without any extracts was added; (6)the control-interdiction group: the Brn-4 polyclonal antibody was added on the basis of the control group. The neurons differentiated from NSCs were detected by MAP-2 immunofluorescence on the 14th day after culturing. Image processing about the number, area and perimeter of MAP-2 positive neurons was carried out respectively. And STATA7.0 statistical software was used to analyze the results. The results showed that the sequence from above the 3 morphological marks was the transection group, the normal group, the transection-interdiction group, the normal-interdiction group, the control group and the control-interdiction group. Statistical results showed that there were significant differences between every two groups (P<0.05) except for between the control group and the control-interdiction group. The present results suggest that Brn-4 polyclonal antibody could blocked partly the role of the fimbria-transected hippocampi extracts inducing NSCs differentiating into neurons in vitro, and Brn-4 may play an important role in the process of NSCs differentiating into neurons.