转录组测序分析猪胚胎附植的中期和后期卵巢差异表达基因

doi:10.11843/j.issn.0366-6964.2018.09.009

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (9): 1879-1888.doi: 10.11843/j.issn.0366-6964.2018.09.009

转录组测序分析猪胚胎附植的中期和后期卵巢差异表达基因

付言峰^1*, 李兰², 赵为民¹, 方晓敏¹, 李碧侠¹, 王学敏¹, 周李生¹, 任守文^1*

1. 江苏省农业科学院畜牧研究所/农业部种养结合重点实验室/江苏省农业种质资源保护与利用平台, 南京 210014;
2. 江苏省农业科学院动物免疫工程研究所, 南京 210014

收稿日期:2017-12-14 出版日期:2018-09-23 发布日期:2018-09-23
通讯作者: 任守文,研究员,E-mail:shouwenren@163.com;付言峰,副研究员,E-mail:fuyanfeng2013@foxmail.com
作者简介:付言峰(1982-),男,山东聊城人,副研究员,博士,主要从事猪遗传育种与繁殖研究,E-mail:fuyanfeng2013@foxmail.com,Tel:025-84391941;李兰(1983-),女,山东高密人,助理研究员,博士,主要从事猪免疫制剂制备技术研究,E-mail:lanli99@foxmail.com
基金资助:
国家科技支撑计划项目（2015BAD03B01-08）；国家生猪现代产业技术体系南京综合试验站（CARS-35）；江苏省农业重大新品种创制项目（PZCZ201733）；江苏省农业科技自主创新资金项目（CX（17）2002）；江苏省科技计划重点项目（BE2017324）；国家自然科学基金项目（31201767）

Differentially Expressed Genes Analysis between Mid-implantation and Post-implantation in Porcine Ovary by RNA Sequencing

FU Yan-feng^1*, LI Lan², ZHAO Wei-min¹, FANG Xiao-min¹, LI Bi-xia¹, WANG Xue-min¹, ZHOU Li-sheng¹, REN Shou-wen^1*

1. Key Laboratory of Crop and Livestock Integrated Farming of Ministry of Agriculture/Jiangsu Germplasm Resources Protection and Utilization Platform, Institute of Animal Science/Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;
2. Institute of Animal Immune Engineering, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China

Received:2017-12-14 Online:2018-09-23 Published:2018-09-23

摘要/Abstract

摘要：

胚胎附植是影响母猪产仔数的一个重要因素，本研究旨在找到猪胚胎附植的关键调控基因。选用5～7胎次梅山猪母猪4头，在其妊娠第18和24天分别屠宰2头，采集其卵巢组织进行转录组测序（RNA-seq）分析，GO功能富集分析及Pathway分析。结果表明：1）猪胚胎附植中期卵巢（卵_18 d）和后期卵巢（卵_24 d）两个组织样检测到的表达基因中最多的序列（reads）均为外显子序列；卵_18 d测得reads最多的染色体依次为6、14、1和7号，卵_24 d依次为1、7、14和M号；卵_24 d相较于卵_18 d有更多的基因表达，且表达量更高；两个时期卵巢组织中表达量前50位的基因中，线粒体基因占60%以上。2）卵_24 d相较于卵_18 d，有581个基因上调和103个基因下调，其中二者之间表达差异最大的基因为EN19684，位于线粒体中；表达差异最大的染色体基因为EN11278，位于14号染色体中。3）差异表达基因显著富集于内皮细胞活化等38个生物学过程，宿主细胞质等19个细胞学分区和FAD-AMP裂解酶活性等16个分子功能；差异表达基因在185条生物通路上存在差异，其中前3位极显著差异的生物通路依次为PI3K-Akt信号通路、Hippo信号通路和昼夜周期。综上表明，在卵巢组织表达的基因，重点参与了胚胎附植后期的调控，且高表达量基因多集中在细胞线粒体中，差异表达基因功能以内皮细胞活化为主，生物通路以PI3K-Akt信号通路为主。

Abstract:

Embryo implantation is one of the important factors affecting litter size in sows. In order to find the key genes regulating porcine embryo implantation. 2 Meishan sows selected and slaughtered on 18^th day and 24^th day of pregnancy,respectively followed by ovary tissues collection. And these tissues would be used in RNA sequencing (RNA-seq) analysis, GO analysis and pathway analysis.The results showed that:1) the most of sequences detected (reads) were gene exons in both tissues (ovary_18 d and ovary_24 d) during embryo implantation in pigs.The most reads were located in chromosome 6 (Chr 6) for ovary_18 d, followed by Chr 14, Chr 1 and Chr 7. For ovary_24 d, the most reads were located in Chr 1, followed by Chr 7, Chr14 and Chr M. In addition, there were more genes expressed and higher expression level in ovary_24 d than that in ovary_18 d. More than 60% genes of top 50 were located in mitochondria according to gene expression level, suggesting that mitochondria genes might play an important role in the regulation of porcine embryo implantation.2)581 genes were up-regulated and 103 genes were down-regulated in ovary_24 d compared with that in ovary_18 d. Among these genes, the expression difference of EN19684 was was highest, which was located in mitochondria, the expression difference of EN11278 was highest, which was located in Chr 14. 3)GO analysis results showed that these differentially expressed genes were significantly enriched in 38 biological processes such as endothelial cell activation, 19 cellular compartments such as host cytoplasm, and 16 molecular functions such as FAD-AMP lyase activity. Pathway analysis results indicated that these differentially expressed genes differed among 185 biological pathways, and the top 3 pathways were PI3K-Akt signaling pathway, Hippo signaling pathway and circadian cycle. In conclusion, genes expressed in ovary are mainly involved in the regulation of post-implantation, and high-expression genes are mostly located in mitochondria. The main function of differentially expressed genes is endothelial cell activation, and the main pathway of differentially expressed genes is PI3K-Akt signaling pathway.

中图分类号:

S828.2

付言峰, 李兰, 赵为民, 方晓敏, 李碧侠, 王学敏, 周李生, 任守文. 转录组测序分析猪胚胎附植的中期和后期卵巢差异表达基因[J]. 畜牧兽医学报, 2018, 49(9): 1879-1888.

FU Yan-feng, LI Lan, ZHAO Wei-min, FANG Xiao-min, LI Bi-xia, WANG Xue-min, ZHOU Li-sheng, REN Shou-wen. Differentially Expressed Genes Analysis between Mid-implantation and Post-implantation in Porcine Ovary by RNA Sequencing[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 1879-1888.

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转录组测序分析猪胚胎附植的中期和后期卵巢差异表达基因

Differentially Expressed Genes Analysis between Mid-implantation and Post-implantation in Porcine Ovary by RNA Sequencing

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