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Melatonin Improves the Quality of In Vitro Produced (IVP) Bovine Embryos: Implications for Blastocyst Development Cryotolerance and Modifications of Relevant Gene Expression

机译:褪黑素提高了体外生产的(IVP)牛胚胎的质量:胚泡发育低温耐受性和相关基因表达的修改的含义。

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摘要

To evaluate the potential effects of melatonin on the kinetics of embryo development and quality of blastocyst during the process of in vitro bovine embryo culture. Bovine cumulus–oocyte complexes (COCs) were fertilized after in vitro maturation. The presumed zygotes were cultured in in vitro culture medium supplemented with or without 10−7 M melatonin. The cleavage rate, 8-cell rate and blastocyst rate were examined to identify the kinetics of embryo development. The hatched blastocyst rate, mortality rate after thawing and the relevant transcript abundance were measured to evaluate the quality of blastocyst. The results showed that melatonin significantly promoted the cleavage rate and 8-cell embryo yield of in vitro produced bovine embryo. In addition, significantly more blastocysts were observed by Day 7 of embryo culture at the presence of melatonin. These results indicated that melatonin accelerated the development of in vitro produced bovine embryos. Following vitrification at Day 7 of embryo culture, melatonin (10−7 M) significantly increased the hatched blastocyst rate from 24 h to 72 h and decreased the mortality rate from 48 h to 72 h after thawing. The presence of melatonin during the embryo culture resulted in a significant increase in the gene expressions of DNMT3A, OCC, CDH1 and decrease in that of AQP3 after thawing. In conclusion, melatonin not only promoted blastocyst yield and accelerated in vitro bovine embryo development, but also improved the quality of blastocysts which was indexed by an elevated cryotolerance and the up-regulated expressions of developmentally important genes.
机译:评估褪黑素对体外牛胚胎培养过程中胚胎发育动力学和胚泡质量的潜在影响。牛卵丘-卵母细胞复合物(COC)在体外成熟后受精。将假定的合子在补充有或没有10 -7 M褪黑激素的体外培养基中进行培养。检查卵裂率,8细胞率和囊胚率,以鉴定胚胎发育的动力学。测量孵化的胚泡率,解冻后的死亡率和相关的转录本丰度,以评价胚泡的质量。结果表明,褪黑激素显着​​提高了体外产生的牛胚胎的裂解率和8细胞胚胎的产量。另外,在褪黑激素存在下,在胚胎培养的第7天观察到明显更多的胚泡。这些结果表明褪黑激素加速了体外产生的牛胚胎的发育。在胚胎培养的第7天进行玻璃化后,褪黑激素(10 −7 M)可使孵化后的胚泡率从24 h显着提高到72 h,融化后的死亡率从48 h降低到72 h。褪黑素在胚胎培养过程中的存在导致解冻后DNMT3A,OCC,CDH1的基因表达显着增加,而AQP3的基因表达下降。总之,褪黑激素不仅能提高胚泡产量并加速体外牛胚胎的发育,而且还能提高胚泡的质量,而胚泡的质量则由耐高温性的提高和发育重要基因的表达上调所指示。

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