Ypel3基因在小鼠胚胎发育中的表达与调控

Expression and Regulation of Ypel3 Gene During Mouse Embryo Development

目的:探讨小鼠胚胎发育过程中Ypel3基因的时空特异性表达与调控,为后续功能研究奠定基础。方法:选取胎龄(E)10.5、12.5、14.5、16.5和18.5 d的小鼠胚胎,利用荧光定量RT-PCR技术研究Ypel3基因mRNA的时序性动态表达谱;采用原位杂交技术观察Ypel3基因mRNA在胚胎发育E11.5和E15.5的空间表达谱;应用定量RT-PCR技术检测表观遗传学修饰对Ypel3基因mRNA表达丰度的影响。结果:定量RT-PCR表明该基因从胚胎发育的早中期开始表达,到出生前表达量呈逐渐升高趋势;原位杂交显示E11.5信号出现在脑和心脏中,E15.5信号在脑、舌、心、肺、胸腺、肝、肾等主要脏器中均有表达;甲基化转移酶抑制剂5-氮胞苷(5-Aza)处理的Neuro-2a(N2a)细胞中,Ypel3的表达水平未产生显著变化,而去乙酰化酶抑制剂4-苯丁酸(4-PBA)处理后该基因表达显著升高,5-Aza和4-PBA联合处理后表达水平进一步升高。结论:Ypel3基因在小鼠胚胎发育各阶段有广泛的表达,提示其具有重要作用,且该基因的表达可能受到组蛋白乙酰化的调控。

Objective： To investigate the developmental-specific expression and regulation of Ypel3 gene.Meth-ods： The temporal expression pattern of Ypel3 at embryo day（E） 10.5,12.5,14.5,16.5,18.5 were studied by flu-orescent quantitative PCR,as well as the spatial expression at E11.5 and E15.5 were studied by in situ hy-bridization.We employed the RT-PCR to investigate the influence of epigenetic modification to the expression level of Ypel3.Results： Real-time quantitative PCR showed that Ypel3 gene was expressed at the early mid-stage of the embryo and gradually increased during embryo development.In situ hybridization analysis verified that Ypel3 gene expression in brain and heart at E11.5,and in brain,tongue,heart,lung,thymus,liver and kidney at E15.5.Ypel3 gene expression abundance remained the same in Neuro-2a cell lines treated by DNA methyltransferase in-hibitor 5-azacytidine（5-Aza）,over-expressed in Neuro-2a treated by histone deacetylase inhibitor 4-phenylbutyric acid（4-PBA） and further over-expressed in Neuro-2a co-treated by 5-Aza and 4-PBA.Conclusion： Ypel3 gene is widely expressed during mouse embryogenesis and may be regulated by histone acetylation.