Algarra Blanca, Maillo Verónica, Avilés Manuel, Gutiérrez-Adán Alfonso, Rizos Dimitrios, Jiménez-Movilla María
Department of Cell Biology and Histology, Faculty of Medicine, Instituto Murciano de Investigación Biosanitaria (IMIB-Arrixaca-UMU), University of Murcia, Murcia 30100, Spain.
Department of Animal Reproduction, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid 28040, Spain.
J Reprod Dev. 2018 Oct 12;64(5):433-443. doi: 10.1262/jrd.2018-058. Epub 2018 Aug 6.
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.
此前,我们的研究小组证明,重组猪输卵管蛋白(pOVGP1)可与体外成熟(IVM)猪卵母细胞的透明带(ZP)结合,对体外受精(IVF)有积极影响。在IVM卵母细胞内检测到pOVGP1这一事实表明,该蛋白在胚胎发育过程中具有生物学作用。本研究的目的是评估pOVGP1对牛体外胚胎发育的影响。我们在IVF、胚胎体外培养(IVC)或两者过程中应用10或50 µg/ml的pOVGP1,以评估卵裂和胚胎发育情况。通过分析重要发育基因的表达和玻璃化/复温后的存活率来评估囊胚质量。在囊胚的ZP、卵周隙和质膜中检测到了pOVGP1。当在IVF或IVC过程中使用10或50 µg/ml的pOVGP1时,在卵裂或囊胚产量方面未发现显著差异(P>0.05)。然而,当在IVF + IVC过程中使用50 µg/ml pOVGP1时,获得的囊胚数量是对照组和10 µg/ml pOVGP1组的一半。用pOVGP1培养的扩张囊胚在玻璃化/复温后的存活率在各组之间没有显著差异(P>0.05)。在IVF、IVC或两者过程中使用pOVGP1可增加胚胎质量标记基因DSC2、ATF4、AQP3和DNMT3A的mRNA相对丰度。总之,在牛胚胎体外培养过程中使用pOVGP1不会影响胚胎发育率,但就特定基因的相对丰度而言,可产生质量更好的胚胎。