RFRP-3对猪卵母细胞体外成熟的影响

doi:10.11843/j.issn.0366-6964.2022.04.013

Abstract

Abstract: The aim of this study was to investigate the effect of RFRP-3 on in vitro maturation of porcine oocyte. In this study, GV stage oocytes from the ovaries of healthy sows collected from slaughterhouses were randomly divided into 3 groups. The maturation rate of porcine oocytes in each group was calculated after incubating with 0, 10^-6 and 10^-8 mol·L^-1 RFRP-3 for 44 h, respectively. In the subsequent experiment, the collected oocytes were randomly divided into 2 groups. Porcine oocytes were cultured in medium with 0 and 10^-8 mol·L^-1 RFRP-3 for 44 h, respectively. The cumulus expansion of porcine oocytes was observed and the cumulus expansion index and maturation rate of oocytes in each group were calculated. The expression of cumulus expansion factors (PTGS2, HAS2, PTX3), oocyte secretory factors (GDF9, BMP15) and cyclin related genes (CCNB1 and CDK1) were detected by qRT-PCR. The contents of MPF and cAMP in oocytes were detected by ELISA kit. The concentrations of progesterone and estrogen were measured by radioimmunoassay. Oocytes in each group were no less than 100, and each test was repeated 3 times. Compared with the control group, the maturation rate of porcine oocytes cultured with 10^-8mol·L^-1 RFRP-3 was extremely significantly reduced (P<0.01). No significant change in cumulus expansion rate and cumulus expansion index were observed(P>0.05), but the expression of cumulus expansion factors (PTGS2, HAS2, PTX3) was extremely significantly decreased (P<0.01). RFRP-3 could extremely significantly reduce MPF content in porcine oocytes (P<0.01), and had no significant effect on cAMP content (P>0.05). After the addition of RFRP-3, the expression levels of GDF9 (P<0.01) and BMP15 (P<0.05) were promoted, while the expression levels of CCNB1 (P<0.05) and CDK1(P<0.05) were inhibited; meanwhile, the concentrations of progesterone and estrogen in medium of experimental groups were extremely significantly decreased (P<0.01). In conclusion, RFRP-3 inhibits in vitro maturation of porcine oocytes by regulating the expression of maturation related factors and cumulus expansion factors and the secretion of steroid hormones. This study will lay a theoretical foundation for elucidating the regulatory effect of RFRP-3 on mammalian oocytes.

Key words: RFRP-3, porcine, oocyte, IVM

CLC Number:

S828.3

JIA Jinmei, FENG Jianhao, YANG Meixia, ZHANG Yufang, ZHANG Hongliang, WANG Shuilian. Effect of RFRP-3 on in Vitro Maturation of Porcine Oocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1133-1142.

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Effect of RFRP-3 on in Vitro Maturation of Porcine Oocytes

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