TUBITAK MRC Genetic Engineering and Biotechnology Institute, Gebze, Turkey.
Mol Reprod Dev. 2012 Sep;79(9):613-25. doi: 10.1002/mrd.22068. Epub 2012 Jul 26.
Vitrification is becoming a preferred method for pre-implantation embryo cryopreservation. The objective of this study was to determine the differentially expressed genes of in vivo- and in vitro-produced bovine embryos after vitrification. In vitro- (IVF) and in vivo-derived (IVV) bovine blastocysts were identified as follows: in vitro-produced fresh (IVF-F), in vitro-produced vitrified (IVF-V), in vivo-derived fresh (IVV-F), in vivo-derived vitrified (IVV-V). The microarray results showed that 53 genes were differentially regulated between IVF and IVV, and 121 genes were differentially regulated between fresh and vitrified blastocysts (P < 0.05). There were 6, 268, 962, and 17 differentially regulated genes between IVF-F × IVV-F, IVF-V × IVV-V, IVF-F × IVF-V, and IVV-F × IVV-V, respectively (P < 0.05). While gene expression was significantly different between fresh and vitrified IVF blastocysts (P < 0.05), it was similar between fresh and vitrified IVV blastocysts. Significantly up-regulated KEGG pathways included ribosome, oxidative phosphorylation, spliceosome, and oocyte meiosis in the fresh IVF blastocyst samples, while sphingolipid and purine metabolisms were up-regulated in the vitrified IVF blastocyst. The results showed that in vitro bovine blastocyst production protocols used in this study caused no major gene expression differences compared to those of in vivo-produced blastocysts. After vitrification, however, in vitro-produced blastocysts showed major gene expression differences compared to in vivo blastocysts. This study suggests that in vitro-produced embryos are of comparable quality to their in vivo counterparts. Vitrification of in vitro blastocysts, on the other hand, causes significant up-regulation of genes that are involved in stress responses.
玻璃化已成为胚胎冻存的首选方法。本研究旨在探讨玻璃化对体内和体外生产的牛胚胎差异表达基因的影响。将体外受精（IVF）和体内生产（IVV）的牛囊胚鉴定如下：体外新鲜生产（IVF-F）、体外玻璃化生产（IVF-V）、体内新鲜生产（IVV-F）、体内玻璃化生产（IVV-V）。微阵列结果显示，IVF 和 IVV 之间有 53 个基因差异表达，新鲜和玻璃化囊胚之间有 121 个基因差异表达（P<0.05）。IVF-F×IVV-F、IVF-V×IVV-V、IVF-F×IVF-V 和 IVV-F×IVV-V 之间分别有 6、268、962 和 17 个差异表达基因（P<0.05）。虽然新鲜和玻璃化 IVF 囊胚之间的基因表达差异显著（P<0.05），但新鲜和玻璃化 IVV 囊胚之间的基因表达相似。显著上调的 KEGG 通路包括核糖体、氧化磷酸化、剪接体和卵母细胞减数分裂，新鲜 IVF 囊胚中鞘脂和嘌呤代谢上调，玻璃化 IVF 囊胚中核糖体、氧化磷酸化、剪接体和卵母细胞减数分裂上调。结果表明，与体内生产的囊胚相比，本研究中使用的体外牛囊胚生产方案没有引起主要的基因表达差异。然而，玻璃化后，体外生产的囊胚与体内囊胚相比，基因表达差异较大。本研究表明，体外生产的胚胎与体内胚胎质量相当。另一方面，体外囊胚的玻璃化会导致参与应激反应的基因显著上调。
