低氧分压对牦牛卵丘细胞转录组的影响  

Effects of Low Oxygen Tension on Transcriptome of Yak(Bos grunniens)Cumulus Cells

作　　者：李瑞哲 徐惊涛[2] 孙永刚[2] 马志杰[2] 陈生梅[2] 罗玉柱[1] LI Rui-Zhe;XU Jing-Tao;SUN Yong-Gang;MA Zhi-Jie;CHEN Sheng-Mei;LUO Yu-Zhu(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;Qinghai Academy of Animal Science and Veterinary Medicine,Xining 810016,China)

机构地区：[1]甘肃农业大学动物科学技术学院,兰州730070 [2]青海省畜牧兽医科学院,西宁810016

出　　处：《农业生物技术学报》2021年第4期688-697,共10页Journal of Agricultural Biotechnology

基　　金：青海省畜牧兽医科学院基本科研业务费(MKY-2019-07);科技部国际合作项目(2013DFA31420)。

摘　　要：卵丘细胞是卵母细胞的重要支持细胞,卵丘细胞的质量影响卵母细胞的体外成熟和发育能力。低氧分压有利于牦牛(Bos grunniens)卵母细胞的体外成熟和发育能力,但是卵丘细胞是否参与该作用尚未见报道。为明确低氧分压对牦牛卵丘细胞转录组的影响,本研究在5%和20%O2的条件下进行牦牛卵丘细胞-卵母细胞复合体(cumulus-oocyte complex,COC)成熟培养,然后收集卵丘细胞进行转录组测序、差异表达基因筛选和qRT-PCR验证。本研究共获得8个转录组文库,总数据量56.17 Gb;基因表达分析表明,两组样品中共有13584个基因得到表达,筛选获得270个差异表达基因,其中229个差异基因在5%O2处理组高表达,41个差异基因在20%O2处理组高表达;基因本体论(Gene Ontology,GO)和KEGG通路富集分析表明,差异表达基因主要富集于糖酵解、卵丘细胞扩散和卵丘细胞-卵母细胞信号传导等生物学过程;在基因组未被注释的区域寻找新基因和新转录本,发现6711个新基因,7209个新转录本,其中829个新转录本具有蛋白编码能力。qRT-PCR结果验证了转录组测序结果准确。上述研究结果提示,低氧分压可能促使牦牛卵丘细胞的转录模式趋同于体内成熟环境,进而促进牦牛卵母细胞的成熟并提高其发育能力。本研究结果为深入研究氧分压对牦牛卵母细胞体外成熟中卵丘细胞-卵母细胞互作机理的影响提供了参考依据。Cumulus cells are important supporting cells of oocytes.Quality of cumulus cells influences in vitro maturation and developmental competence of oocytes.Low oxygen tension was beneficial for in vitro maturation and developmental competence of yak(Bos grunniens)oocytes,whereas whether the beneficial effects were exerted through cumulus cells has not been reported.In order to investigate the effects of low oxygen tension on transcriptome of yak cumulus cells,maturation of yak cumulus-oocyte complex(COC)was carried out at 5%and 20%O2,respectively.Then cumulus cells were collected for RNA-seq,screening of differentially expressed genes(DEGs)and q RT-PCR verification.The results showed that 8 yak cumulus cell transcriptomic libraries were obtained and total data size was 56.17 Gb;gene expression analysis showed that a total of 13584 genes were expressed of the two groups and 270 DEGs were identified,among which 229 DEGs were up-regulated in 5%O2 and 41 DEGs were up-regulated in 20%O2;Gene Ontology(GO)and KEGG pathway analysis showed that DEGs were mostly enriched in glycolysis,cumulus cell expansion and cumulus-oocyte signaling et al.;6711 new genes and 7209 new transcripts,among which 829 new transcripts had protein coding potential,were found in unannotated region of the yak reference genome.qRT-PCR results were consistent with the sequencing data.Taken together,above results indicated that low oxygen tension might induce yak cumulus cells to have a similar transcriptomic profile to those matured in vivo and therefore promote the maturation and developmental competence of yak oocytes.The present study provides a reference for further study of the effect of oxygen tension on the mechanism of cumulus cell-oocyte interaction in the process of in vitro maturation of yak oocytes.

关 键 词：牦牛 卵丘细胞 转录组 氧分压 

分 类 号：S814[农业科学—畜牧学]