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目的检测小鼠心脏发育过程中nesprin-1基因mRNA和蛋白的表达及亚细胞定位。方法取小鼠胚胎12.5 d(embryo 12.5day,E12.5)、E14.5、E16.5、E18.5以及新生鼠、成年小鼠心脏,应用RT-PCR、Western blot、免疫荧光技术检测胚胎各时间点、新生鼠、成年鼠心脏nesprin-1基因mRNA和蛋白表达及细胞定位。结果在小鼠胚胎E12.5、E14.5、E16.5、E18.5以及新生鼠、成年小鼠心脏组织中,nesprin-1基因mRNA及蛋白均有表达,其表达量随着心脏的发育而逐渐增加,E16.5和E18.5表达最高,成年鼠表达最低。免疫荧光显示,nesprin-1的分布在各个时期均位于核被膜及核周间隙中。结论nesprin-1在小鼠心脏发育过程中表达呈动态变化,在心脏传导系统和心肌细胞发育成熟时期呈高表达,因此推测nesprin-1可能在心脏传导系统和心肌细胞的发育中有重要作用。
Objective To detect the expression and subcellular localization of nesprin-1 mRNA and protein during cardiac development in mice. Methods The embryos were harvested at embryonic day 12.5, embryonic 12.5 days, E14.5, E16.5 and E18.5, and from neonatal and adult mouse hearts. The embryos were detected by RT-PCR, Western blot and immunofluorescence At each time point, nesprin-1 mRNA and protein expression and cellular localization were detected in neonatal and adult rat hearts. Results The mRNA and protein of nesprin-1 were expressed in mouse embryos E12.5, E14.5, E16.5, E18.5 and neonatal rat and adult mouse hearts respectively. But gradually increased, the expression of E16.5 and E18.5 was the highest, and the expression of E16.5 and E18.5 was the lowest. Immunofluorescence showed that the distribution of nesprin-1 was located in the nuclear envelope and perinuclear space at all stages. Conclusion The expression of nesprin-1 changes dynamically during heart development in mice and is highly expressed during the development of cardiac conduction and cardiomyocyte. Therefore, nesprin-1 may play an important role in the development of cardiac conduction and cardiomyocyte.