Effect of Ovarian Extract on Oocyte Maturation and Early Embryonic Development in Pigs

S. Yang, Jae-Hun Choi, Young-Seo Jo, Ye-Won Kim, Dong-Mok Lee, Hyo-Jin Park, D. Koo
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Abstract

Various factors in the ovary are known to regulate oocyte maturation and hormone secretory functions; however, the effect of ovarian extract (OE) on oocyte maturation and embryonic development in pigs remains unknown. In this study, we first evaluated whether OE supplementation in the in vitro maturation (IVM) medium alters the oocyte maturation capacity by affecting glucose/amino acid metabolites, meiotic maturation, cumulus cell (CC) expansion, and antioxidants. Various OE concentrations (50, 100, 200, 500, and 5000 μg/mL) were included in the IVM medium. Only the oocytes treated with 100 μg/mL OE exhibited an improved meiotic maturation rate when compared with that of the other groups (non-treated group, 78.6 ± 3.0% vs. 100 μg/mL OE-treated group, 81.6 ± 4.3%); however, the difference was not significant. To observe the changes in glucose and amino acid metabolism in the OE-treated oocytes, we measured the amounts of diverse constituents (glucose, lactate, glutamine, and ammonia) in the IVM medium containing OE. Lactate and ammonia levels in the OE-treated group after 44 h of IVM were higher (p < 0.01) than those in the non-treated group. In addition, the expression of the CC expansion factors (Has2 and Tnfaip6) significantly increased (p < 0.05), whereas the mRNA expression levels of antioxidative enzymes (Sod1, Cat, and Gpx1) significantly diminished (p < 0.05) in the OE-treated group. Moreover, mature oocytes treated with 100 μg/mL OE demonstrated increased subsequent embryonic development rates after 144 h of IVM. Thus, the addition of OE in IVM mediums may improve oocyte maturation capacity which could enhance antioxidative enzyme activation, energy metabolism, and expression of the CC expansion factors in porcine oocytes.
卵巢提取物对猪卵母细胞成熟和早期胚胎发育的影响
卵巢中有多种因素调节卵母细胞成熟和激素分泌功能;然而,卵巢提取物(OE)对猪卵母细胞成熟和胚胎发育的影响尚不清楚。在这项研究中,我们首先评估了在体外成熟(IVM)培养基中添加OE是否会通过影响葡萄糖/氨基酸代谢物、减数分裂成熟、积云细胞(CC)扩张和抗氧化剂来改变卵母细胞的成熟能力。在IVM培养基中加入不同浓度的OE(50、100、200、500、5000 μg/mL)。与其他各组相比,只有100 μg/mL OE处理的卵母细胞减数分裂成熟率有所提高(未处理组为78.6±3.0%,100 μg/mL OE处理组为81.6±4.3%);然而,差异并不显著。为了观察OE处理的卵母细胞中葡萄糖和氨基酸代谢的变化,我们测量了含有OE的IVM培养基中不同成分(葡萄糖、乳酸、谷氨酰胺和氨)的含量。IVM 44 h后,oe处理组乳酸和氨水平显著高于未处理组(p < 0.01)。此外,oe组CC扩张因子(Has2、Tnfaip6)表达量显著升高(p < 0.05),而抗氧化酶(Sod1、Cat、Gpx1) mRNA表达量显著降低(p < 0.05)。此外,100 μg/mL OE处理的成熟卵母细胞在IVM 144 h后的胚胎发育率增加。因此,在IVM培养基中添加OE可以提高猪卵母细胞的成熟能力,从而增强猪卵母细胞抗氧化酶的激活、能量代谢和CC扩增因子的表达。
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