梅山猪胚胎附植期EphA1基因表达及甲基化检测

Detection of EphA1 Expression and Methylation during Embryo Implantation in Meishan Pigs

旨在研究促红细胞生成素产生肝细胞受体A1(EphA1)在某些哺乳动物胚胎发育期的细胞间侵入和黏附活动中发挥一定作用以及EphA1基因甲基化对胚胎附植期其表达的影响。本研究以梅山猪为研究对象,采用实时荧光定量PCR、Western blot和亚硫酸氢盐测序法,分析了EphA1基因的mRNA表达、蛋白表达和CpG岛甲基化。结果表明,在梅山猪胚胎附植前期(13d)、中期(18d)、后期(24d)的子宫内膜附植点,EphA1的mRNA表达趋势为先升高后降低(P<0.05),蛋白表达趋势为逐渐降低,且妊娠猪的表达量均显著高于空怀猪(P<0.05)。附植前期的EphA1甲基化程度最高,中期的次之,后期的最小,即EphA1甲基化趋势与蛋白表达趋势一致。综上表明,EphA1的CpG岛甲基化很可能影响了基因的表达,进一步影响了猪胚胎附植的调控。

Erythropoietin-producing hepatocellular receptor and ligand A1(EphA1)plays a role in cell migration and adhesion during embryonic development in various mammals.The purpose of the study is to analyze EphA1 methylation effect on its expression during embryo implantation.In this study,the mRNA expression,protein expression and CpG island methylation of EphA1 was investigated in Meishan pigs using real-time quantitative PCR,Western blot and bisulfate sequencing technology.The results showed that EphA1 mRNA expression increased from 13 to 18dand decreased from 18 to 24d(P<0.05)in endometrium implantation sites of Meishan pigs,while EphA1 protein expression decreased from 13 to 24d,and expression in pregnant sows was significantly higher than that of non-pregnant sows(P<0.05)during pre-implantation(13d),mid-implantation(18d)and post-implantation(24d)period.About CpG island methylation level of EphA1,it had a highest methylation pre-implantation period,followed by mid-implantation andpost-implantation,this indicated EphA1 CpG island methylation level was consistent with protein expression level during pre-implantation,mid-implantation and post-implantation period.These findings suggest that CpG island methylation of EphA1 might affect its expression and further affect the regulation of swine embryo implantation.