抑制UCHL1对猪卵母细胞体外成熟、透明带泛素化及多精入卵的影响

doi:10.16431/j.cnki.1671-7236.2017.12.024

Abstract

Abstract:

This study was aimed to examine the effects of UCHL1 inhibition on porcine oocyte maturation in vitro, zona pellucida (ZP) ubiquitination and polyspermy. DAPI staining, Hoechst staining and SDS-PAGE methods were used to detect the matuation rate of porcine oocytes in vitro, the level of ubiquitination of zona pellucida (ZP) and polyspermy. The results showed that after different concentrations UCHL1 inhibitor (10, 20, 25 and 30 μmol/L, DMSO and control group) were added into maturation medium for culturing 46 h in vitro, the mature rate of control group was 86.22%, while the maturation rate of 30 μmol/L group was 15.30%, and the maturation rate of every treatment group had significant difference (P<0.05). Western blotting result showed that every group generated ubiquitin markers of ZP were about 61, 80 and 106 ku in different degree. According to the analysis of gray value, the result had significant difference (P<0.05). Conducting fertilization in vitro, the number of sperm adhered on oocyte ZP in control group was the most, the number of sperm running into oocyte was fewer, the number of sperm adhered on oocyte ZP with 30 μmol/L UCHL1 inhibitor was the fewest, and there was almost no sperm running into the oocyte. The results showed that UCHL1 inhibitor had an impact on maturation of porcine oocytes in vitro. With the higher concentration of UCHL1, the lower degree of ZP protein ubiquitinated, UCHL1 could regulate sperm attachment and polyspermy.

Key words: porcine oocyte; UCHL1 inhibitor; IVM; ZP ubiquitination; polyspermy

CLC Number:

S814

SHI Wen-shu, JIN Yi. Effects of UCHL1 Inhibition on Porcine Oocyte Matuation in vitro, Zona Pellucida Ubiquitination and Polyspermy[J]. , 2017, 44(12): 3563-3569.

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Effects of UCHL1 Inhibition on Porcine Oocyte Matuation in vitro, Zona Pellucida Ubiquitination and Polyspermy

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