Complexin蛋白对神经母瘤细胞分化的影响及机制研究  

The Effects and Mechanisms of Complexin on the Differentiation of Murine Neuroblastoma

作　　者：龚吉红 蒋伟[1] 李小红[1] 林显光[1] 阳小飞[1] 

机构地区：[1]中南民族大学生物医学工程学院脑认知国家民委重点实验室、膜离子通道与药物研发实验室,生命科学学院,湖北武汉430074

出　　处：《现代生物医学进展》2016年第16期3032-3035,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(31300892);湖北省自然科学基金项目(2014CFA027,2014CFB455)

摘　　要：目的:探讨Complexin蛋白对神经母瘤细胞分化的影响及机制。方法:采用神经母瘤细胞(N2a)作为实验材料,在其中过表达Complexin蛋白以及其突变体后,利用激光共聚焦显微镜拍照并利用Image J软件对N2a细胞的分化比率、突起数量以及突起生长长度进行统计学分析。结果:在N2a细胞中过表达Cpx蛋白后,细胞分化率比对照组(即转染空白对照质粒)增加约2倍。Cpx1-86和Cpxpoorclamp突变体可促进N2a细胞的分化,而Cpx27-134突变体对N2a细胞分化无明显影响。随着时间的延长,过表达野生型Cpx蛋白和其N端缺失突变体都不能显著增加细胞突起的数量;但在转染4天后,过表达野生型Cpx蛋白能显著增加分化细胞的突起长度,而其N端缺失突变体不能引起突起长度的增加。结论:Complexin蛋白主要通过其N端序列促进神经母瘤细胞(N2a)的分化,增加分化后突起的长度,但对突起数量没有明显影响。Objective： To investigate the effects and mechanisms of Complexin on the differentiation of murine neuroblastoma. Methods： Complexin and its mutants were overexpressed by using murine neuroblastoma （N2a cells） as experiment materials. The pictures of N2a cells were taken by laser scanning confocal microscope, and analyzed by using Image J to determine the cell differentiation ratio, branch number and neurite length. Results： The differential rate of N2a cells was increased two times after Cpx overexpression compared to the control group （transfected with blank vector） approximately. The N2a cells differentiation could be promoted by the mutants of Cpx^1-86 and Cpx^poorclamp but not by the mutant of Cpx^27-134. No obvious increase in the number of neurite was observed in either the wild-type Cpx or its N-terminal deletion mutant over-expressed N2a cells, regardless of the experimental period. The length of neurite was significantly increased by over-expressing the wild-type but not the N-terminal deletion mutant of Cpx after 4 days transfection. Conclusion： Complexin increased the rate of N2a cells differentiation and the length of neurite, but not the number of neurite. Moreover, these phenotypes were mainly depended on the N-terminal sequence of complexin.

关 键 词：神经细胞分化 COMPLEXIN N2A细胞 神经递质释放 

分 类 号：R-33[医药卫生] Q189[生物学—神经生物学]