梅山猪胚胎附植期EphB2的组织表达及RNA-seq分析

doi:10.3724/SP.J.1005.2014.1243

遗传 ›› 2014, Vol. 36 ›› Issue (12): 1243-1248.doi: 10.3724/SP.J.1005.2014.1243

梅山猪胚胎附植期EphB2的组织表达及RNA-seq分析

付言峰¹, 周艳红^{1, 2}, 王爱国³, 李兰^{4, 5}, 刘红林², 李碧侠¹, 任守文¹

1. 江苏省农业科学院畜牧研究所, 动物品种改良和繁育重点实验室, 南京 210014;
2. 南京农业大学动物科技学院, 南京 210095;
3. 中国农业大学动物科学技术学院, 北京 100193;
4. 中国农业大学动物医学院, 北京 100193;
5. 国家兽用生物制品工程技术研究中心, 南京 210014

收稿日期:2014-03-22 出版日期:2014-12-20 发布日期:2014-12-20
通讯作者: 任守文, 研究员, 硕士生导师, 研究方向：动物遗传育种与生产。E-mail: shouwenren@163.com E-mail:fuyanfeng@jaas.ac.cn;954268869@qq.com
作者简介:付言峰, 博士, 副研究员, 专业方向：动物遗传育种与繁殖。Tel: 025-84391941;周艳红, 在读硕士研究生, 专业方向：动物遗传育种与繁殖。付言峰和周艳红同为第一作者。
基金资助:
国家自然科学基金项目(编号：31201767), 江苏省农业科技自主创新资金项目(编号：CX(13)5042), 国家生猪现代产业技术体系-南京综合试验站项目(编号：nycytx-009)和江苏省科技支撑计划项目(编号：BE2012333)资助

Tissue expression of EphB2 and RNA-seq analysis during embryo im-plantation in Meishan pigs

Yanfeng Fu¹, Yanhong Zhou^{1, 2}, Aiguo Wang³, Lan Li^{4, 5}, Honglin Liu², Bixia Li¹, Shouwen Ren¹

1. The Key Laboratory of Animal Breed Improvement and Reproduction, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;
2. College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China3. College of Animal Science and Technology, China Agricultural University, Beijing 100193, China;
4. College of Veterinary Medicine, China Agricultural University, Beijing 100193, China;
5. National Research Center of Veterinary Biologicals Engineering and Technology, Nanjing 210014, China

Received:2014-03-22 Online:2014-12-20 Published:2014-12-20

摘要/Abstract

摘要： 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。

关键词: EphB2, 胚胎附植, 表达, 转录组测序, 梅山猪

Abstract: Embryo implantation is a key step affecting swine litter size, which is an important economic and reproduction 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 expression 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 implantation 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 followed 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.

Key words: EphB2, embryo implantation, expression, RNA-seq, Meishan pig

付言峰, 周艳红, 王爱国, 李兰, 刘红林, 李碧侠, 任守文. 梅山猪胚胎附植期EphB2的组织表达及RNA-seq分析[J]. 遗传, 2014, 36(12): 1243-1248.

Yanfeng Fu, Yanhong Zhou, Aiguo Wang, Lan Li, Honglin Liu, Bixia Li, Shouwen Ren. Tissue expression of EphB2 and RNA-seq analysis during embryo im-plantation in Meishan pigs[J]. HEREDITAS(Beijing), 2014, 36(12): 1243-1248.

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梅山猪胚胎附植期EphB2的组织表达及RNA-seq分析

Tissue expression of EphB2 and RNA-seq analysis during embryo im-plantation in Meishan pigs

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