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Effects of recombinant OVGP1 protein on in vitro bovine embryo development

机译:重组OVGP1蛋白对体外牛胚胎发育的影响

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摘要

Previously, our group demonstrated that recombinant porcine oviductin (pOVGP1) binds to the zona pellucida (ZP) of in vitro-matured (IVM) porcine oocytes with a positive effect on in vitro fertilization (IVF). The fact that pOVGP1 was detected inside IVM oocytes suggested that this protein had a biological role during embryo development. The aim of this study was to evaluate the effects of pOVGP1 on bovine in vitro embryo development. We applied 10 or 50 µg/ml of pOVGP1 during IVF, embryonic in vitro culture (IVC), or both, to evaluate cleavage and embryo development. Blastocyst quality was assessed by analyzing the expression of important developmental genes and the survival rates after vitrification/warming. pOVGP1 was detected in the ZP, perivitelline space, and plasma membrane of blastocysts. No significant differences (P > 0.05) were found in cleavage or blastocyst yield when 10 or 50 µg/ml of pOVGP1 was used during IVF or IVC. However, when 50 µg/ml pOVGP1 was used during IVF + IVC, the number of blastocysts obtained was half that obtained with the control and 10 µg/ml pOVGP1 groups. The survival rates after vitrification/warming of expanded blastocysts cultured with pOVGP1 showed no significant differences between groups (P > 0.05). The use of pOVGP1 during IVF, IVC, or both, increased the relative abundance of mRNA of DSC2, ATF4, AQP3, and DNMT3A, the marker-genes of embryo quality. In conclusion, the use of pOVGP1 during bovine embryo in vitro culture does not affect embryo developmental rates but produces embryos of better quality in terms of the relative abundance of specific genes.
机译:先前,我们的小组证明重组猪输卵管蛋白(pOVGP1)与体外成熟(IVM)猪卵母细胞的透明带(ZP)结合,对体外受精(IVF)具有积极作用。在IVM卵母细胞中检测到pOVGP1的事实表明,该蛋白在胚胎发育过程中具有生物学作用。这项研究的目的是评估pOVGP1对牛体外胚胎发育的影响。我们在体外受精,胚胎体外培养(IVC)或两者中应用10或50 µg / ml的pOVGP1来评估卵裂和胚胎发育。通过分析重要发育基因的表达和玻璃化/变暖后的存活率来评估胚泡质量。在囊胚的ZP,玻璃体周间隙和质膜中检测到pOVGP1。在IVF或IVC期间使用10或50μg/ ml的pOVGP1时,卵裂或囊胚产率没有显着差异(P> 0.05)。但是,在IVF + IVC期间使用50 µg / ml pOVGP1时,获得的胚泡数是对照组和10 µg / ml pOVGP1组的胚泡数的一半。在玻璃化/加热后,用pOVGP1培养的膨胀胚泡的存活率在各组之间无显着差异(P> 0.05)。在IVF,IVC或两者同时使用pOVGP1可以增加DSC2,ATF4,AQP3和DNMT3A(胚胎品质的标记基因)mRNA的相对丰度。总之,在牛胚胎体外培养过程中使用pOVGP1不会影响胚胎的发育速度,但是就特定基因的相对丰度而言,可以生产出质量更高的胚胎。

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