China Animal Husbandry and Veterinary Medicine ›› 2021, Vol. 48 ›› Issue (9): 3387-3393.doi: 10.16431/j.cnki.1671-7236.2021.09.030

• Genetics and Breeding • Previous Articles     Next Articles

Study on the Expression and Subcellular Localization of G Protein-coupled Receptor 50 During in vitro Maturation Process of Yak Oocytes

YAO Ying1, CHEN Yan1, XIONG Xianrong1,2, MIPAM Tserang-donko3, CHAI Zhixin3, JI Wenhui1, LAN Daoliang1,2   

  1. 1. College of Animal & Veterianry Sciences, Southwest Minzu University, Chengdu 610041, China;
    2. Key Laboratory of Qinghai-Tibet Plateau Animal Genetic Resource and Utilization, Ministry of Education, Chengdu 610041, China;
    3. Institute of Qinghai-Tibetan Plateau, Southwest Minzu University, Chengdu 610041, China
  • Received:2021-01-21 Online:2021-09-20 Published:2021-09-17

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Abstract: This study was aimed to explore the site of G protein-coupled receptor 50 (GPR50) expression and localization in yak oocytes during in vitro maturation (IVM), and to provide the basis for better understanding the molecular mechanism of oocyte maturation and the specificity of yak reproduction. After yak oocytes were matured in vitro, fluorescence staining was used to monitor the changes of spindle morphology and nuclear phase at different time points (0-24 h). The time points of the four critical periods of maturity for yak oocytes, including germinal vesicle(GV), germinal vesicle break down(GVBD), metaphase Ⅰ(M Ⅰ) and metaphase Ⅱ(M Ⅱ) were determined accurately. On this basis, the dynamic expression of GPR50 gene during the maturation of yak oocytes was detected by Real-time quantitative PCR, and the subcellular dynamic site of GPR50 protein during the maturation of oocytes was detected by immunofluorescence staining. The results showed that 90% of the in vitro matured yaks oocytes were at GV stage at 0 h, 94% at GVBD stage at 6 h, 92% at MⅠ stage at 16 h, 94% at MⅡ stage at 24 h. The results of Real-time quantitative PCR showed that GPR50 gene was expressed in GV phase of yak oocytes, and it gradually increased during the maturation of GVBD, MⅠ and MⅡ stages, and reached the peak at MⅡ stage, which was extremely significantly higher than GV and GVBD stages (P<0.01). GPR50 protein was mainly expressed on the membrane at GV stage, and then dispersed to cytoplasm gradually with the development of maturation, and high brightness dispersion expression was found at MⅡ stage. The above results indicated that GPR50 gene might play an important role in the meiosis of yak oocytes, and provided for the study on the function and mechanism of GPR50 in yak oocytes.

Key words: G protein-coupled receptor 50 (GPR50); yak; oocytes; meiosis

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