猪MII期卵母细胞冷冻前后全基因组表达谱分析  

Whole genome expression profile analysis of vitrified porcine MII stage oocytes

作　　者：戴建军[1,2,3] 吴彩凤 张树山[1,2,3] 孙玲伟[1,2] 陈亚宁 张德福 DAI Jian-jun;WU Cai-feng;ZHANG Shu-shan;SUN Ling-wei;CHEN Ya-ning;ZHANG De-fu(Institute of Animal Science and Veterinary Medicine,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China;Division of Animal Genetic Engineering,Shanghai Municipal Key Laboratoryof Agri-genetics and Breeding,Shanghai 201106,China;Shanghai Engineering Research Center ofBreeding Pig,Shanghai 201106,China)

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海201106 [2]上海市农业遗传育种重点实验室动物遗传工程研究室,上海201106 [3]上海种猪工程技术研究中心,上海201302

出　　处：《上海农业学报》2019年第2期67-74,共8页Acta Agriculturae Shanghai

基　　金：国家自然科学基金(31372315);国家转基因生物新品种培育科技重大专项(2014ZX08006-005);上海市科委工程技术研究中心建设专项(16DZ2250700);上海市农业科学院攀高计划(PG171)

摘　　要：为探究猪MII期卵母细胞冷冻前后基因表达的变化,采用猪全基因组表达谱芯片对其冷冻前后的基因进行表达谱分析,筛选出2倍以上差异表达基因,并利用生物信息学对这些基因进行GO和KEGG Pathway分析。结果表明:(1)猪卵母细胞玻璃化冷冻前后共获得2倍以上差异表达基因171个,其中冻后上调表达基因103个,下调表达基因68个;选取的6个差异表达基因的RT-PCR验证结果与芯片分析结果一致。(2)损伤应答、细胞凋亡、程序性细胞死亡、细胞坏死及蛋白激酶调节、RNA拼接、核mRNA拼接和mRNA代谢等参与了卵母细胞冻后基因差异表达的生物学过程;(3)胞外区、基底质膜、细胞外间隙、质膜、肌动蛋白细胞骨架、线粒体脊、线粒体、细胞器膜和线粒体包膜等参与了卵母细胞冻后基因差异表达的细胞组分调控;(4)类固醇结合、酶抑制剂的活性、激素结合、核苷酸结合、辅酶结合和ATP酶活等参与了卵母细胞冻后差异表达基因的分子功能调控。(5) KEGG Pathway分析表明,Toll样受体信号通路、NOD样受体信号通路、黏着连接和MAPK信号通路等参与了冻后卵母细胞的基因表达调控。综上所述,冷冻能造成猪MII期卵母细胞多系统的损伤,如线粒体损伤、凋亡、坏死、质膜损伤和RNA拼接与代谢异常等。In order to find out the main changes of gene expression levels of porcine oocytes before and after vitrification,this study conducted the whole genome expression profiling with pig whole-genome microarrays,screened over-2-fold differentially expressed genes,and analyzed those genes by GO and KEGG pathway.The results showed that:1)compared with non-vitrified oocytes,there were 171 over-2-fold differentially expressed genes,including 103 up-regulated genes and 68 down-regulated genes;2)responses to wounding,regulation of apoptosis,regulation of programmed cell death,regulation of cell death,regulation of protein kinase,RNA splicing,nuclear mRNA splicing and mRNA metabolism were involved in the biological process of differentially expressed genes after GO analysis;3)extracellular region part,basolateral plasma membrane,extracellular space,plasma membrane part,actin cytoskeleton,mitochondrial part,mitochondrion,organelle membrane and mitochondrial envelope were involved in the cellular component of differentially expressed genes after GO analysis;4)steroid binding,enzyme inhibitor activity,hormone binding,coenzyme binding and ATPase activity were involved in the molecular function of differentially expressed genes after GO analysis;5)after KEGG pathway analysis,Toll-like receptor signaling pathway,NOD-like receptor signaling,adhering junction and MAPK signaling pathway were involved in regulation of gene expression of porcine vitrified oocytes.In conclusion,vitrification could cause the multiple system damages of porcine MII stage oocytes,such as mitochondrial damage,apoptosis,necrosis,plasma membrane damage and abnormal RNA splicing and abnormal metabolism.

关 键 词：猪 卵母细胞 玻璃化冷冻 表达谱 基因芯片 

分 类 号：S828[农业科学—畜牧学]