CX43 and oxidative stress are the targets of BCB staining to predict the developmental potential of buffalo oocytes

Author:

Li MengQi12ORCID,Yang ChunYan13,Duan AnQin13,Xiao Peng124,Lu XingRong13,Ma XiaoYa13,Xu YuanYuan1,Zheng Wei13,Feng Chao13,Mo Xia13,Huang ChenQian13,Huang LiQing1,Shang JiangHua13,Zheng HaiYing13

Affiliation:

1. Guangxi Key Laboratory of Buffalo Genetics, Reproduction and Breeding Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences Nanning China

2. State Key Laboratory for Conservation and Utilization of Subtropical Agro‐Bioresources College of Animal Science and Technology, Guangxi University Nanning Guangxi China

3. Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology Ministry of Agriculture and Rural Affairs Nanning China

4. College of Animal Science and Technology Guangxi Vocational University of Agriculture Nanning China

Abstract

AbstractThis study used the brilliant cresyl blue (BCB) staining method to group buffalo oocytes (BCB+ and BCB−) and perform in vitro maturation, in vitro fertilization and embryo culture. At the same time, molecular biology techniques were used to detect gap junction protein expression and oxidative stress‐related indicators to explore the molecular mechanism of BCB staining to predict oocyte developmental potential. The techniques of buffalo oocytes to analyse their developmental potential and used immunofluorescence staining to detect the expression level of CX43 protein, DCFH‐DA probe staining to detect ROS levels and qPCR to detect the expression levels of the antioxidant‐related genes SOD2 and GPX1. Our results showed that the in vitro maturation rate, embryo cleavage rate and blastocyst rate of buffalo oocytes in the BCB+ group were significantly higher than those in the BCB− group and the control group (p < .05). The expression level of CX43 protein in the BCB+ group was higher than that in the BCB− group both before and after maturation (p < .05). The intensity of ROS in the BCB+ group was significantly lower than that in the BCB− group (p < .05), and the expression levels of the antioxidant‐related genes SOD2 and GPX1 in the BCB+ group were significantly higher than those in the BCB− group (p < .05). Brilliant cresyl blue staining could effectively predict the developmental potential of buffalo oocytes. The results of BCB staining were positively correlated with the expression of gap junction protein and antioxidant‐related genes and negatively correlated with the reactive oxygen species level, suggesting that the mechanism of BCB staining in predicting the developmental potential of buffalo oocytes might be closely related to antioxidant activity.

Funder

Natural Science Foundation of Guangxi Zhuang Autonomous Region

Publisher

Wiley

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