Effects Of Different Treatments On The Zona Pellucida Protein Ubiquitination Of Porcine Oocytes

The purpose of the research is to explore the level of zona pellucida protein ubiquitination in the process of vitro maturation, in the period of porcine oocytes’GV and MⅡ stage, the research is to explore how the PSMD4 affects zona pellucida protein ubiquitination and the function of zona pellucida protein ubiquitination in sperm-oocyte binding. In order to reduce polysperm and provide a theoretical basis and reference on improving sperm fertility. Thus, by understanding the biological characteristics of porcine oocytes, optimize the vitro maturation of porcine oocytes, in vitro fertilization, and the other assisted reproductive technology. SDS-PAGE and Western Blot were used to analyze the zona pellucida protein samples of porcine oocytes with different treatment, the result is as follows:1、The results of zona pellucida protein Western Blot in GV and M Ⅱ stage shows that the 65 kDa、85 kDa and 120 kDa has different ubiqutination level, the grey value of ubiqutination strip is different significantly. The result indicates that zona pellucida protein has different degrees of ubiquitination in vitro maturation of porcine oocytes.2、The Western blot result of adding PSMD4 antibodies in maturation medium aiming at porcine zona pellucida protein shows that ZP1 has ubiqutination on the 120 kDa with the group without adding PSMD4 antibodies and ZP1 has’t ubiqutination with the group adding PSMD4 antibodies accordingly. There are different degrees of ubiquitination with the two group on 65 kDa and 85 kDa. The grey value of ubiqutination strip is different of two groups. The result shows that ZP1 has’t ubiquitination, and the ubiquitintion level decrease of ZP2 and ZP3.3、The results of Western blot shows that the 65kDa,85kDa and 120kDa zona pellucida protein has different ubiqutination in different culticating time and all the zona pellucida ubiquitination level decreased after 52 h. The results indicates that the oocyte ages and the free ubiquitin decreases in the process of vitro maturation, resulting the level decrease of ubiquitintion.