Wnt-1对室管膜前下区神经干细胞体外分化的影响  

Effects of Wnt-1 on the differentiation of anterior subventricular zone neural stem cells in vitro

作　　者：邱克军[1] 杨辉[1] 蔡文琴[2] 姚忠祥[3] 何家全[1] 刘仕勇[1] 吕胜青[1] 

机构地区：[1]第三军医大学新桥医院神经外科,重庆400037 [2]第三军医大学新桥医院神经生物教研室,重庆400037 [3]第三军医大学新桥医院组胚教研室,重庆400037

出　　处：《中华神经外科杂志》2005年第7期403-407,共5页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金重点资助项目(30130110)

摘　　要：目的探讨室管膜前下区(anteriorsubventricularzone,SVZa)神经干细胞体外分化过程中,Nestin和Wnt-1表达的变化,以及Wnt-1对SVZa神经干细胞体外分化的影响。方法以免疫荧光染色方法观察在SVZa神经干细胞体外分化过程中,Nestin和Wnt-1在不同时间点的表达水平,同时以基因转染方法增强SVZa神经干细胞Wnt-1表达,观察Nestin、Wnt-1和Mash-1的表达以及神经元分化比例的改变。结果⑴Nestin主要表达于SVZa神经干细胞分化的早期阶段,随着细胞的分化和成熟,Nestin的表达迅速减弱。⑵SVZa神经干细胞贴壁分化2h即有Wnt-1表达,其表达高峰在细胞分化开始后12h,以后随着细胞分化的进行而迅速降低。⑶SVZa神经干细胞转染Wnt-1基因后,Nestin的表达减弱,Mash-1的表达增强,神经元的分化比例增高,Mash-1的增强与神经元分化之间有很高的一致性。结论⑴Wnt-1表达于SVZa神经干细胞分化过程之中,在细胞分化成熟后则不表达,Wnt-1可能参与调节SVZa神经干细胞的体外分化,并可作为神经干细胞处于分化状态的标志。⑵SVZa神经干细胞转染Wnt-1后表达Mash-1增强,并更多分化为神经元,Wnt-1可能通过Mash-1途径促使SVZa神经干细胞向神经元方向分化。Objective To explore the expression pattern of Nestin and Wnt-1 during anterior subventricular zone (SVZa) neural stem cells (NSCs) differentiating in vitro, also to study the effects of Wnt-1 on the neuronal differentiation of SVZa NSCs. Methods The expression levels of Nestin and Wnt-1 were observed on different time points by immunofluorescence during SVZa NSCs differentiating in vitro. The expressions of Nestin, Wnt-1, Mash-1 and the neuronal differentiation of SVZa NSCs were also studied while Wnt-1 eukaryocyte expression plasmid transfected. Results ⑴ Nestin expressed mainly in the early stage of SVZa NSCs differentiation, and decreased rapidly with SVZa NSCs differentiating and maturating. ⑵ Wnt-1 expressed throughout the course of SVZa NSCs differentiation. The highest expression of Wnt-1 was at 12th hour after SVZa NSCs differentiation, then it decreased with SVZa NSCs differentiating and maturating. ⑶ Transfected with Wnt-1, the expression of Wnt-1 and Mash-1, an important factor involved in neuron differentiation, enhanced, but the expression of Nestin, a marker of NSCs, reduced significantly. In the same time, the number of neurons differentiated from SVZa NSCs was increased. Conclusions It is possible to conclude that Wnt-1 is essential for the differentiation of SVZa NSCs in vitro and may be a marker that NSCs are differentiating. Enhancing the expression of Wnt-1 may suppress the stem cell properties of SVZa NSCs and promote SVZa NSCs to differentiate into neurons through Mash-1 signal pathway.

关 键 词：WNT-1 细胞体外分化 室管膜 Nestin SVZA 神经干细胞分化 神经元分化 分化过程 细胞转染 染色方法 免疫荧光 不同时间 基因转染 早期阶段 细胞贴壁 表达增强 1基因 一致性 多分化 比例 成熟 减弱 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]