Laboratory of Developmental Biology, Institute of Animal Physiology and Genetics of the Czech Academy of Sciences, 27721 Liběchov, Czech Republic.
Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, 94901 Nitra, Slovakia.
Int J Mol Sci. 2021 Oct 15;22(20):11148. doi: 10.3390/ijms222011148.
The developmental potential of porcine oocytes cultured in vitro was remarkably enhanced in a medium containing FGF2, LIF and IGF1 (FLI) when compared to a medium supplemented with gonadotropins and EGF (control). We analyzed the molecular background of the enhanced oocyte quality by comparing the time course of MAPK3/1 and AKT activation, and the expression of genes controlled by these kinases in cumulus-oocyte complexes (COCs) cultured in FLI and the control medium. The pattern of MAPK3/1 activation in COCs was very similar in both media, except for a robust increase in MAPK3/1 phosphorylation during the first hour of culture in the FLI medium. The COCs cultured in the FLI medium exhibited significantly higher activity of AKT than in the control medium from the beginning up to 16 h of culture; afterwards a deregulation of AKT activity occurred in the FLI medium, which was not observed in the control medium. The expression of cumulus cell genes controlled by both kinases was also modulated in the FLI medium, and in particular the genes related to cumulus-expansion, signaling, apoptosis, antioxidants, cell-to-cell communication, proliferation, and translation were significantly overexpressed. Collectively, these data indicate that both MAPK3/1 and AKT are implicated in the enhanced quality of oocytes cultured in FLI medium.
与添加促性腺激素和表皮生长因子的培养基(对照)相比,在含有 FGF2、LIF 和 IGF1(FLI)的培养基中培养的猪卵母细胞的体外发育潜能显著提高。我们通过比较 MAPK3/1 和 AKT 激活的时间过程以及在 FLI 和对照培养基中培养的卵丘-卵母细胞复合物(COC)中受这些激酶控制的基因的表达,分析了增强卵母细胞质量的分子背景。在两种培养基中,COC 中 MAPK3/1 的激活模式非常相似,除了在 FLI 培养基中培养的第一个小时内 MAPK3/1 磷酸化的强烈增加。从培养开始到 16 小时,在 FLI 培养基中 AKT 的活性明显高于对照培养基;之后,在 FLI 培养基中观察到 AKT 活性的失调,而在对照培养基中没有观察到。两种激酶控制的卵丘细胞基因的表达也在 FLI 培养基中受到调节,特别是与卵丘扩展、信号转导、细胞凋亡、抗氧化剂、细胞间通讯、增殖和翻译相关的基因显著过表达。总的来说,这些数据表明,MAPK3/1 和 AKT 都参与了 FLI 培养基中培养的卵母细胞质量的提高。