Kim Jin-Woo, Park Hyo-Jin, Yang Seul-Gi, Kim Min-Ji, Kim In-Su, Jegal Ho-Geun, Wee Gabbine, Yang Hee-Young, Park Joung Jun, Choo Young-Kug, Koo Deog-Bon
Department of Biotechnology, College of Engineering, Daegu University, 201 Daegudae-ro, Jillyang, Gyeongsan, Gyeongbuk, 38453, Republic of Korea.
Laboratory Animal Center, Daegu-Gyeongbuk Medical Innovation Foundation (DGMIF), 80 Cheombok-ro, Dong-gu, Daegu, 41061, Republic of Korea.
Reprod Sci. 2020 Jan;27(1):278-289. doi: 10.1007/s43032-019-00004-9. Epub 2020 Jan 1.
Ganglioside GT1b is well-known for its role in cytokine production and in activating epidermal growth factor receptor (EGFR)-mediated signaling pathways in cancer cells. However, there are no reports that clearly elucidate the role of GT1b in EGFR-mediated signaling pathways in porcine oocytes during the process of in vitro maturation (IVM). In this study, we investigated the role of GT1b in EGFR-mediated activation of the ERK1/2 pathway in porcine cumulus-oocyte complexes (COCs) at 44 h of IVM. Our data show that expression of the ST3GAL2 protein significantly increased in porcine COCs at 44 h irrespective of treatment with EGF. Meiotic maturation and mRNA levels of factors (HAS2, TNFAIP6, and PTX3) related to cumulus cell expansion significantly increased in COCs treated with 2 μM GT1b during IVM in the absence of EGF. They also increased in COCs treated with EGF/GT1b as compared to that in the other groups. Interestingly, protein levels of EGFR, phospho-EGFR, ERK1/2, and phospho-ERK1/2 dramatically increased in COCs treated with EGF/GT1b. Moreover, the rate of fertilization and the developmental competence of blastocyst were significantly higher in EGF/GT1b-treated COCs. Taken together, these results suggest that exogenous GT1b improves meiotic maturation and cumulus cell expansion in porcine COCs via activation of EGFR-mediated ERK1/2 signaling.
神经节苷脂GT1b因其在细胞因子产生以及激活癌细胞中表皮生长因子受体(EGFR)介导的信号通路中的作用而闻名。然而,尚无报道明确阐明GT1b在猪卵母细胞体外成熟(IVM)过程中EGFR介导的信号通路中的作用。在本研究中,我们研究了GT1b在IVM 44小时时对猪卵丘-卵母细胞复合体(COCs)中EGFR介导的ERK1/2通路激活的作用。我们的数据表明,无论是否用表皮生长因子(EGF)处理,ST3GAL2蛋白的表达在IVM 44小时的猪COCs中均显著增加。在无EGF的IVM期间,用2 μM GT1b处理的COCs中,与卵丘细胞扩张相关的因子(HAS2、TNFAIP6和PTX3)的减数分裂成熟和mRNA水平显著增加。与其他组相比,用EGF/GT1b处理的COCs中这些指标也有所增加。有趣的是,用EGF/GT1b处理的COCs中EGFR、磷酸化EGFR、ERK1/2和磷酸化ERK1/2的蛋白水平显著增加。此外,用EGF/GT1b处理的COCs的受精率和囊胚发育能力显著更高。综上所述,这些结果表明,外源性GT1b通过激活EGFR介导的ERK1/2信号通路改善猪COCs的减数分裂成熟和卵丘细胞扩张。