Plate-forme de Chirurgie et d'Imagerie pour la Recherche et l'Enseignement (CIRE), Pôle d'Analyse et d'Imagerie des Biomolécules (PAIB), INRA, CHRU de Tours, Université de Tours, Nouzilly, France.
UMR PRC, INRA 85, CNRS 7247, University of Tours, IFCE, Nouzilly, France.
Mol Reprod Dev. 2019 Jun;86(6):661-672. doi: 10.1002/mrd.23144. Epub 2019 Apr 4.
The objective of this study was to evaluate the effect of progesterone (P4), estradiol (E2), and cortisol (CO) at intraoviductal concentrations on bovine embryo development and quality in vitro. After fertilization of in vitro matured oocytes, zygotes were cultured for 8 days in synthetic oviductal fluid, supplemented with 55 ng/ml P4, 120 pg/ml E2, 40 ng/ml CO, or their combination (ALL). Control embryos were cultured with vehicle (0.1% ethanol). Exposure to steroids did not affect the embryo developmental rate nor the mean number of cells per blastocyst. However, at 24 hr after vitrification-warming, exposure to P4 improved the proportion of embryos that re-expanded and were viable while exposure to CO decreased the proportion of viable embryos. By intact cell MALDI-TOF mass spectrometry, a total of 242 phospholipid masses of 400-1000 m/z were detected from individual fresh blastocysts. Exposure to ALL induced the highest and most specific changes in embryo phospholipids, followed by P4, E2, and CO. In particular, the m/z 546.3 and 546.4 attributed to lysophosphatidylcholines were found less abundant after exposure to P4. In conclusion, exposure of bovine embryos to intraoviductal concentrations of steroid hormones did not affect in vitro development but changed blastocyst quality in terms of cryotolerance and phospholipid profiles.
本研究旨在评估孕激素(P4)、雌二醇(E2)和皮质醇(CO)在输卵管内浓度对牛胚胎体外发育和质量的影响。体外成熟卵母细胞受精后,胚胎在含有 55ng/ml P4、120pg/ml E2、40ng/ml CO 或其组合(ALL)的合成输卵管液中培养 8 天。对照胚胎用载体(0.1%乙醇)培养。类固醇暴露不影响胚胎发育率或每个囊胚的细胞平均数。然而,在玻璃化-解冻后 24 小时,暴露于 P4 可提高胚胎重新扩张和存活的比例,而暴露于 CO 则降低存活胚胎的比例。通过完整细胞 MALDI-TOF 质谱法,从单个新鲜囊胚中检测到 400-1000m/z 的 242 种磷脂质质量。ALL 暴露诱导胚胎磷脂质发生最高和最特异的变化,其次是 P4、E2 和 CO。特别是,暴露于 P4 后,归因于溶血磷脂酰胆碱的 m/z 546.3 和 546.4 丰度降低。总之,牛胚胎暴露于输卵管内类固醇激素浓度不影响体外发育,但在冷冻耐受性和磷脂谱方面改变了囊胚质量。