CAJ | 학술논문

猪产仔数是一个重要的经济和繁殖性状，而胚胎附植是影响猪产仔数的重要因素。为了研究促红细胞生成素产生肝细胞受体B2(EphB2)对猪胚胎附植过程中子宫内膜迁移和粘附活动的影响，文章以太湖流域梅山猪为研究对象，利用实时荧光定量PCR (qRT-PCR)和蛋白免疫印迹(Western blot)方法，检测EphB2基因在梅山猪胚胎附植前、中、后期子宫内膜附植点/非附植点和卵巢组织的mRNA和蛋白表达谱，并用转录组测序(RNA-seq)方法分析了不同附植时期子宫内膜附植点和卵巢组织的差异表达基因。qRT-PCR 和 Western blot 结果表明， EphB2在胚胎附植前、中、后期子宫内膜附植点和非附植点的mRNA和蛋白均呈现先升高后降低的表达趋势，且附植中期的表达量显著高于前期和后期(P<0.01)；EphB2在附植前、中、后期卵巢中的mRNA和蛋白表达趋势则相反，为先降低后升高，且不同时期间表达差异显著(P<0.05)。RNA-seq 结果表明，EphB2在子宫内膜附植点的 mRNA 表达，附植中期极显著高于附植前期(P<0.01)；EphB2在卵巢的 mRNA 表达，附植中期显著高于附植后期(P<0.05)。综上所述，EphB2很可能在猪胚胎附植过程中发挥着重要的调控作用，为潜在的猪产仔数性状候选基因。
저산자수시일개중요적경제화번식성상，이배태부식시영향저산자수적중요인소。위료연구촉홍세포생성소산생간세포수체B2(EphB2)대저배태부식과정중자궁내막천이화점부활동적영향，문장이태호류역매산저위연구대상，이용실시형광정량PCR (qRT-PCR)화단백면역인적(Western blot)방법，검측EphB2기인재매산저배태부식전、중、후기자궁내막부식점/비부식점화란소조직적mRNA화단백표체보，병용전록조측서(RNA-seq)방법분석료불동부식시기자궁내막부식점화란소조직적차이표체기인。qRT-PCR 화 Western blot 결과표명， EphB2재배태부식전、중、후기자궁내막부식점화비부식점적mRNA화단백균정현선승고후강저적표체추세，차부식중기적표체량현저고우전기화후기(P<0.01)；EphB2재부식전、중、후기란소중적mRNA화단백표체추세칙상반，위선강저후승고，차불동시기간표체차이현저(P<0.05)。RNA-seq 결과표명，EphB2재자궁내막부식점적 mRNA 표체，부식중기겁현저고우부식전기(P<0.01)；EphB2재란소적 mRNA 표체，부식중기현저고우부식후기(P<0.05)。종상소술，EphB2흔가능재저배태부식과정중발휘착중요적조공작용，위잠재적저산자수성상후선기인。
Embryo implantation is a key step affecting swine litter size, which is an important economic and re-production trait in pigs. In order to investigate the effect of erythropoietin-producing hepatocellular receptor B2 (EphB2) on endometrium migration and attachment during swine embryo implantation, the mRNA and protein ex-pression levels of EphB2 in endometrium implantation sites, endometrium non-implantation sites and ovary were detected in Meishan sows during pre-implantation, mid-implantation and post-implantation period using real-time quantitative PCR and Western blot. Differential expression genes were also analyzed in endometrium implantation sites and ovary during different implantation periods by RNA sequencing (RNA-seq) technology. The qRT-PCR and Western blot results showed that EphB2 mRNA and protein expression curve was the same in endomtrium implanta-tion sites and endometrium non-implantation sites during pre-implantation, mid-implantation and post-implantation period, with a first increase followed by a decrease, and its expression level during mid-implantation was significantly higher than pre-implantation and post-implantation (P<0.01). In contrast, EphB2 mRNA and protein expression curve in ovary during pre-implantation, mid-implantation and post-implantation period showed a first decrease fol-lowed by an increase, and the expression levels were significantly different among different implantation periods (P<0.05). RNA-seq results indicated that EphB2 mRNA expression during mid-implantation was higher than that of pre-implantation extremely significantly in endometrium implantation sites (P<0.01), and was significantly higher than that of post-implantation in ovary (P<0.05). By and large, EphB2 might play an important role in swine embryo implantation, and it’s a potential candidate gene for litter size in pigs.