基于RNA-Seq分析氨气对肉兔卵丘卵母细胞复合体的影响  

Effect of ammonia on cumulus oocyte complex of meat rabbit based on RNA-Seq

作　　者：崔嘉[1] 吴峰洋 杨新宇[1] 樊长林 陈宝江[1,3] CUI Jia;WU Fengyang;YANG Xinyu;FAN Changlin;CHEN Baojiang(College of Animal Science and Technology,Hebei Agricultural University,Baoding,Hebei 071000,China;Baoding Animal Quarantine Team,Baoding,Hebei 071001,China;Hebei Technology Innovation Center of Cattle and Sheep Embryo,Baoding,Hebei 071000,China)

机构地区：[1]河北农业大学动物科技学院,河北保定071000 [2]保定市动物检疫队,河北保定071001 [3]河北省牛羊胚胎技术创新中心,河北保定071000

出　　处：《中国兽医学报》2021年第11期2280-2287,共8页Chinese Journal of Veterinary Science

基　　金：国家重点研发计划资助项目(2016YFD0500501-7)。

摘　　要：本试验旨在研究氨气对肉兔卵丘卵母细胞复合体(COCs)的影响。选取100只35日龄雌性肉兔于环境控制仓内,控制氨气浓度分别为0.0 mg/m^(3)(CG组)与7.6 mg/m^(3)(LAC组),试验期28 d。使用苏木精-伊红染色法(HE)检测卵巢组织结构完整性,并采用高通量转录组测序RNA-Seq技术对肉兔COCs进行检测,对检测结果进行拼接与质量评估,筛选出差异表达基因,并进行GO和KEGG功能分析,同时用荧光定量PCR对5个与卵泡发育有关的差异表达基因进行验证。结果表明,氨气处理后肉兔卵巢组织结构完整性被破坏,与对照组(CG组)相比,LAC组原始卵泡与初级卵泡的数量极显著降低(P<0.01),闭锁卵泡增多(P>0.05)。CG与LAC组的差异表达基因(differently expression genes,DEGs)数量为3345个,其中显著上调DEGs 1093个,显著下调DEGs 2252个,其中发现多个与繁殖相关的高表达基因,如SIRT1、KIRREL、IGFBP2、Pik3r1、KDM2B和LHCGR等基因。结合GO富集分析,发现差异表达基因主要富集于49个条目,这些基因主要富集在DNA转录调控、细胞质、核、胞质溶胶成分及各种物质的结合方面。KEGG通路分析表明,差异基因主要参与卵母细胞减数分裂与孕激素介导的卵母细胞成熟通路,以及参与细胞增殖和分化、激素调节功能的PI3K-Akt、Rap1、Hippo、FoxO和TGF-beta信号通路等31个信号通路。本试验为进一步阐明氨气影响肉兔的生殖调控提供了依据。To investigate the effect of ammonia on the cumulus-oocyte complexes(COCs)of rabbits.35-day-old female rabbits were selected in the environmental control warehouse,and the ammonia concentration was controlled to 0 ppm(CG)and 10 ppm(LAC).The test period was 28 d.Hematoxylinand eosin staining(HE)was used to detect the integrity of ovarian tissue,and high-throughput transcriptome sequencing(RNA-Seq)technology was used to detect the effect of ammonia on COCs.The differentially expressed genes were screened out and the GO enrichment analysis and KEGG pathway analysis were performed.Five differentially expressed genes related to follicular development were identified by fluorescence quantitative PCR.Compared with the control group,the number of primordial follicles and primary follicles in LAC group decreased significantly(P<0.01),and atresia follicles increased(P>0.05).The number of differentially expressed genes(DEGs)in CG and LAC groups were 3345,of which 1093 were significantly up-regulated and 2252 were significantly down-regulated.Several highly expressed genes related to reproduction were found,such as SIRT1,KIRREL,IGFBP2,Pik3 r1,KDM2 B and LHCGR.Combined with GO enrichment analysis,it was found that the DEGs were mainly concentrated in 49 items,which were mainly concentrated in DNA transcription regulation,cytoplasm,nucleus,cytoplasmic lysis components and the binding of various substances.KEGG pathway analysis showed that the DEGs were mainly involved in meiosis and progesterone-mediated oocyte maturation,and 31 signaling pathways including PI3 K Akt,Rap1,Hippo,FOXO and TGF-beta signaling pathways involved in cell proliferation and differentiation and hormone regulation.This study provides a basis for elucidating the effect of ammonia on reproductive regulation of meat rabbits.

关 键 词：S814氨气 肉兔 卵丘卵母细胞复合体 转录组 

分 类 号：S856.9[农业科学—临床兽医学]