Saenz-de-Juano M D, Peñaranda D S, Marco-Jiménez F, Vicente J S
Instituto de Ciencia y Tecnología Animal, Universidad Politécnica de Valencia, 46022 Valencia, Spain.
Instituto de Ciencia y Tecnología Animal, Universidad Politécnica de Valencia, 46022 Valencia, Spain.
Cryobiology. 2014 Aug;69(1):178-80. doi: 10.1016/j.cryobiol.2014.06.002. Epub 2014 Jun 12.
Vitrification is replacing slow freezing as the most popular method for oocyte and embryo cryopreservation. However, very little information is available on alterations in epigenetic regulation. Previous studies reported post-implantation effects of vitrification on fetal development and gene expression. This study was conducted to determine if vitrification procedure induce alterations in OCT4 promoter methylation profile which could determine the set point of fetal losses and transcriptomic alterations observed after implantation. Rabbit morulae were recovered at Day 3 of development and vitrified and transferred, or directly transfer, to recipient till Day 6. A conserved regulation region of OCT4 promoter was examined in control and vitrified embryos by bisulfite sequencing and quantitative PCR was used to measure the gene expression. No significant differences were observed in methylation levels or gene expression of OCT4. This work was the first approach in rabbit to the study of possible epigenetic alterations associated with vitrification procedure.
玻璃化冷冻正取代慢速冷冻，成为卵母细胞和胚胎冷冻保存最常用的方法。然而，关于表观遗传调控改变的信息却非常少。先前的研究报道了玻璃化冷冻对植入后胎儿发育和基因表达的影响。本研究旨在确定玻璃化冷冻过程是否会诱导OCT4启动子甲基化谱的改变，而这种改变可能决定植入后观察到的胎儿丢失和转录组改变的设定点。在发育第3天收集兔桑葚胚，进行玻璃化冷冻后移植，或直接移植到受体直至第6天。通过亚硫酸氢盐测序检测对照胚胎和玻璃化冷冻胚胎中OCT4启动子的保守调控区域，并使用定量PCR测量基因表达。未观察到OCT4甲基化水平或基因表达的显著差异。这项工作是在兔中首次研究与玻璃化冷冻过程相关的可能表观遗传改变。
