Acta Veterinaria et Zootechnica Sinica ›› 2022, Vol. 53 ›› Issue (11): 3856-3865.doi: 10.11843/j.issn.0366-6964.2022.11.012

• ANIMAL BIOTECHNOLOGY AND REPRODUCTION • Previous Articles     Next Articles

Effect of Altered O-GlcNAc Modification on in vitro Maturation of Bovine Oocytes

WANG Tengfei, FENG Zhiqiang, SUN Yawen, ZHAO Shanjiang, HAO Haisheng, ZOU Huiying, DU Weihua, ZHAO Xueming, ZHU Huabin, PANG Yunwei*   

  1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Received:2022-03-28 Online:2022-11-23 Published:2022-11-25

Abstract: The aim of this study was to explore the effect of altered O-GlcNAc modification on in vitro maturation (IVM) of bovine oocytes. Bovine in vitro-matured(IVM) oocytes were used to detect the distribution of OGT, OGA and O-GlcNAc proteins. Then cumulus-oocyte complexes were matured in IVM medium supplemented with 4 mmol·L-1 OGT inhibitor BADGP or 100 μmol·L-1 OGA inhibitor PUGNAc, respectively. The none supplementation group was used as control. The first polar body extrusion rate and subsequent embryonic development in vitro were detected, the O-GlcNAc level as well as the mRNA and protein expression levels of OGT, OGA, GFAT and TXNIP were detected by RT-qPCR and Western blot. Results showed that OGT and O-GlcNAc proteins were colocalized in the cytoplasm and nucleus of oocytes, while OGA and O-GlcNAc proteins were colocalized in the cytoplasm and primarily enriched at the cortex. Compared with control group, the first polar body extrusion rate ((52.8±5.1)% & (60.9±1.9)% vs. (70.8±5.4)%) and the blastocyst formation rate ((9.6±4.9)% & (10.0±5.8)% vs. (21.5±4.3)%) significantly reduced in BADGP and PUGNAc groups (P<0.05). BADGP treatment significantly reduced the expression of OGT, the level of O-GlcNAc modification in oocytes was down-regulated, and the expression of OGA was decreased; In the PUGNAc-treated group, the expression of OGA was significantly down-regulated, the modification level of O-GlcNAc was increased, and the protein expression of OGT was significantly decreased. Inhibition of OGT significantly up-regulated the expression of GFAT mRNA, a key rate-limiting enzyme in the hexosamine biosynthesis pathway, whereas inhibition of OGA significantly down-regulated the expression of GFAT mRNA. In addition, the altered level of O-GlcNAc modification significantly up-regulated the expression of TXNIP, a key factor in glucose regulation. The results indicated that changes in the level of O-GlcNAc modification would reduce the maturation and developmental ability of bovine oocytes in vitro, and oocytes responded to the fluctuation of O-GlcNAc modification levels by feedback-regulating the expressions of OGT, OGA, GFAT and TXNIP.

Key words: bovine, oocyte, O-GlcNAc modification, OGT, OGA

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