DAT1基因在大鼠神经干细胞分化中作用的初步研究  

Effects of DAT1 gene on differentiation of neural stem cells in rats

作　　者：惠玲[1] 刘建军[1] 姚忠祥[1] 蔡文琴[2] 

机构地区：[1]第三军医大学基础医学部组织学与胚胎学教研室,重庆400038 [2]第三军医大学基础医学部神经生物学教研室,重庆市神经科学研究所,重庆400038

出　　处：《第三军医大学学报》2005年第7期588-591,共4页Journal of Third Military Medical University

基　　金：国家自然科学青年基金资助项目 ( 30 4 0 0 12 5)~~

摘　　要：目的　研究DAT1转染后不同诱导条件下DAT1在神经干细胞的表达以及DAT1过表达对神经干细胞分化的影响。方法　构建真核表达载体pEGFP C1 DAT1、pcDNA4/hisA DAT1,采用lipofectamine2 0 0 0脂质体介导法转染培养的胚胎大鼠皮层神经干细胞(neuralstemcells ,NSCs) ,免疫组化染色,荧光显微镜观察。结果　DAT1瞬时转染神经干细胞后的诱导分化实验表明,DAT1转染可使NSCs分化为Mash 1阳性的细胞增加,DAT1的高表达有促进NSCs向神经元方向分化的作用;血清诱导分化及自然分化后DAT1在分化细胞中的表达定位发生了变化,同时这种变化也反映在NSCs分化的不同阶段。结论　不同的外界诱导因素或在NSCs分化的不同时期,DAT1可能通过与不同的转录因子的结合,从而在不同的信号通路中影响NSCs的分化。Objective To observe DAT1 expression in neural stem cells (NSCs) and the effects of DAT1 on the differentiation of NSCs. Methods Eukaryotic expression vectors pEGFP-C1-DAT1 and pcDNA4/hisA-DAT1 were constructed and transfected into NSCs of rat cerebellum by lipofectamine2000. The transfected NSCs were observed by immunohistochemistry under fluorescent microscope. Results The overexpression of DAT1 could increase the number of Mash-1 staining cells and promote the NSCs to differentiate into neuron progenitors, and the high levels of DAT1 in NSCs facilitated the differentiation of neurons. The localizations of DAT1 protein in Mash-1 staining cells and NF 200 staining cells changed. This shift may result from the two distinct inducing factors, FBS and nature differentiation, or distinct stages in differentiation of NSCs. Conclusion DAT1 functions in differentiation of NSCs as a multiprotein combined with distinct transcription factors by virtue of different inducer or varied stage of differentiation.

关 键 词：DAT1 神经干细胞 分化 大鼠 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]