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Inhibition of PSMD 4 alters ZP 1 ubiquitination state and sperm–oocyte‐binding ability in pigs
Author(s) -
Xuan B,
Li ZC,
Wang QY,
Xu M,
Chen X,
Jin Y
Publication year - 2018
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.13158
Subject(s) - oocyte , ubiquitin , zona pellucida , andrology , human fertilization , sperm , in vitro , biology , microbiology and biotechnology , chemistry , embryo , genetics , medicine , gene
Contents The aim of this study was to determine how the duration of culture affects the ubiquitination of zona pellucida ( ZP ) proteins ( ZP 1, ZP 2 and ZP 3) during porcine oocyte maturation in vitro. We analysed the changes in ZP protein ubiquitination under three conditions: (i) during oocyte maturation from stage GV to MII ; (ii) in oocytes cultured for different periods of time; and (iii) in oocytes treated with an antibody against PSMD 4. Our results show that ZP 1 and ZP 2 are ubiquitinated at the GV stage, while ZP 1, ZP 2 and ZP 3 are ubiquitinated at the MII stage, and band intensities for these proteins were significantly different between the GV and MII stages ( p < .05). We also found that ubiquitination occurs in ZP 1, ZP 2 and ZP 3 after cultured for 46, 52, 58 and 64 hr, and that the level of ubiquitinated ZP 1 was significantly different in oocytes that were cultured for different time periods. Finally, treatment with an antibody against PSMD 4 resulted in a significant decrease in ZP 1 ubiquitination ( p < .05), without affecting ZP 2 or ZP 3. The number of attached sperms per oocyte was also significantly different between control and anti‐ PSMD 4‐treated groups. Thus, we concluded that ZP 1 and ZP 2 are ubiquitinated at the GV stage, and ZP 1, ZP 2 and ZP 3 are ubiquitinated at the MII stage. As the duration of culture increases, the ubiquitination levels of ZP proteins decrease. We also found that PSMD 4 improves ZP 1 ubiquitination during in vitro culture of porcine oocytes and effectively inhibits sperm–oocyte binding.