nesprins基因在胚胎干细胞分化为心肌细胞过程中的表达  

Study on expression of nesprins gene during mouse embryonic stem cells differentiation into cardiomyocytes in vitro

作　　者：王芳洁[1] 易岂建[1] 郭鹏飞[1] 刘晓燕[1] 张秋萍[2] Shanahan C 

机构地区：[1]重庆医科大学附属儿童医院心脏中心,重庆400014 [2]剑桥大学心血管实验室,英国剑桥

出　　处：《重庆医科大学学报》2010年第5期652-656,共5页Journal of Chongqing Medical University

基　　金：高等学校博士学科点专项科研基金(教技发中心函[2005]216号文20050631005);留学回国人员科研启动基金(教外司留[2007]24号文)

摘　　要：目的:探讨nesprins基因在小鼠胚胎干细胞(Embryonic stem cell,ESC)分化为心肌细胞过程中的表达及作用。方法:体外培养小鼠ESC,采用5-氮杂胞苷为诱导剂诱导ESC分化为心肌细胞,并进行心肌细胞鉴定;RT-PCR检测分化过程中各时间点(d7、d7+3、d7+6、d7+9、d7+20)nesprin-1、nesprin-2基因mRNA的表达;免疫荧光检测各时间点nesprin-1蛋白的细胞定位。结果:nesprin-1、nesprin-2基因mRNA在ESC分化为心肌细胞过程中均有表达;nesprin-1基因mRNA在d7、d7+3、d7+6均表达较高,无显著性差异,随后表达呈下降趋势,在d7+20时表达最低(P<0.05);nesprin-2基因mRNA在d7时表达最高,与d7+3、d7+6组间均有显著性差异(P<0.05),且随着分化时间的延长表达逐渐下降,在d7+20时表达最低;nesprin-1蛋白在ESC分化过程中均有表达,定位于核被膜及核周间隙。结论:nesprin-1、nesprin-2基因表达高峰在生心细胞的诱导和特化时期,推测nesprins对维持心肌细胞发育,使ESC更完全的向心肌细胞分化起重要作用。Objective：To investigate expression and role of nesprins gene during mouse embryonic stem cell（ESC）differentiation into cardiomyocyte in vitro.Methods：Mouse ESC cultrued in combining 5-azacytidine as inducer with hanging drop cultures and the differentiated cells were identified with cardiac special genes and proteins;The mRNA expression of nesprin-1 and nesprin-2 genes were detected by reverse transcription-polymerase chain reaction（RT-PCR）,and sub-cellular location of nesprin-1 protein in nucleus was detected by immunofluorescence in d7,d7＋3,d7＋6,d7＋9 and d7＋20.Results：The expression of nesprin-1 and nesprin-2 genes mRNA can be found in the period of mouse ESC differentiation into cardiomyocyte.There is expression peak of nesprin-1 gene mRNA in d7,d7＋3,d7＋6,then declined gradually to the lowest in d7＋20;expression peak of nesprin-2 gene mRNA in d7,then also declined gradually to the lowest in d7＋20;The protein of nesprin-1 can be detected in the period of mouse ESC differentiation into cardiomyocyte,and mainly located in nuclear envelope and space around nuclei.Conclusion：The expression peak of nesprin-1 and nesprin-2 gene mRNA is interrelated with the phase of inducing and specialization of ESC differentiation into cardiac muscle cell,it can be speculated that nesprin genes has an important role in maintaining ESC differentiation into cardiac muscle cell.

关 键 词：NESPRINS 胚胎干细胞 心肌细胞 分化 小鼠 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学] R394-33[医药卫生—基础医学]