抑制泛素结合酶改变猪卵母细胞透明带泛素化状态及精-卵结合能力  

Inhibiton of Ubiquitin-conjugating Enzyme Alters the Ubiquitination Status of Porcine Oocyte Zona Pellucidas and Sperm-oocyte Binding Ability

作　　者：徐妲[1] 王洋[1] 玄彪 汪秋月 徐曼[1] 陈璇[1] 金一[1] XU Da;WANG Yang;XUAN Biao;WANG Qiuyue;XU Man;CHEN Xuan;JIN Yi(Agricultural College of Yanbian University,Yanji 133002,China)

机构地区：[1]延边大学农学院,延吉133002

出　　处：《畜牧兽医学报》2019年第3期544-550,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31260529);吉林省科技厅重点科技攻关项目(20150204079NY)

摘　　要：本研究旨在探究猪卵母细胞体外成熟过程中添加泛素结合酶(E2)抑制剂对卵母细胞透明带蛋白泛素化水平及精-卵结合能力的影响。试验分为6组:对照组、DMSO组、5、10、15和20μmol·L^(-1) NSC697923处理组。采用Western blot方法分析体外成熟培养液中添加不同浓度泛素结合酶抑制剂NSC697923对猪卵母细胞透明带泛素化水平表达的影响。通过Hoechst染色检测不同组别的精卵结合能力。结果表明:1)猪卵母细胞体外成熟培养液中,添加10、15和20μmol·L^(-1) NSC697923显著降低卵母细胞成熟率(P<0.05),透明带硬化时间(P<0.05)以及精卵结合率(P<0.05)。2)对照组和各处理组的透明带蛋白在61、81、106 ku处发生不同程度的泛素化标记,而添加15和20μmol·L^(-1) NSC697923显著地降低透明带蛋白泛素化水平(P<0.05)。综上表明,猪卵母细胞体外成熟过程中泛素结合酶(E2)改变成熟卵母细胞透明带泛素化水平以及精卵结合能力。The main purpose of this study was to investigate the influence of the adding of ubiquitin-conjugating enzyme(E2) inhibitors to oocyte zona pellucida(ZP)ubiquitination and sperm- oocyte binding ability in vitro maturation of porcine oocytes. There was 6 groups in this study:control group,DMSO,5,10,15 and 20 μmol·L -1 NSC697923 treatment groups. We analyzed the effect of different concentrations of ubiquitin-conjugating enzyme inhibitor NSC697923 on the expression of ubiquitination in porcine oocytes ZP with Western blot in vitro maturation. The difference of sperm-oocyte binding ability among different groups was examined by Hoechst staining. The result showed that:1) In vitro maturation of porcine oocytes, the treatment groups with 10,15 and 20 μmol·L -1 of NSC697923C significantly reduced oocyte maturation rate ( P <0.05), ZP hardening time ( P <0.05) and the sperms-oocyte rate ( P <0.05). 2) The ZP proteins in the control group and each treatment groups showed different degrees of ubiquitination at 61, 81 and 106 ku, while the treatment group with 15 and 20 μmol·L -1 of NSC697923 significantly reduced the ZP ubiquitination protein level ( P <0.05).In conclusion, ubiquitin-conjugating enzyme changes the level of ubiquitination of mature oocytes ZP and the ability of sperm-oocyte binding in vitro maturation of porcine oocytes.

关 键 词：猪 卵母细胞 泛素结合酶 透明带泛素化 精子-卵子结合 

分 类 号：S828.2[农业科学—畜牧学]