ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2019, Vol. 50 ›› Issue (3): 544-550.doi: 10.11843/j.issn.0366-6964.2019.03.009

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

XU Da, WANG Yang, XUAN Biao, WANG Qiuyue, XU Man, CHEN Xuan, JIN Yi*   

  1. Agricultural College of Yanbian University, Yanji 133002, China
  • Received:2018-05-18 Online:2019-03-23 Published:2019-03-23

Abstract:

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.

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