Labrecque Rémi, Sirard Marc-André
Centre de Recherche en Biologie de la Reproduction, Université Laval, Québec, QC, Canada.
Reprod Fertil Dev. 2011;23(4):591-602. doi: 10.1071/RD10243.
The processes underlying the very first moments of embryonic development are still not well characterised in mammals. To better define the kinetics of events taking place following fertilisation, it would be best to have perfect synchronisation of sperm entry. With fertilisation occurring during a time interval of 6 to 12h in the same group of fertilised oocytes, this causes a major variation in the time of activation of embryonic development. Bovine parthenogenesis could potentially result in better synchronisation and, if so, would offer a better model for studying developmental competence. In the present study, bovine oocytes were either parthenogenetically activated or fertilised and cultured in vitro for 7 days. Gene expression analysis for those two groups of embryos at early and expanded stages was performed with BlueChip, a customised 2000-cDNA array developed in our laboratory and enriched in clones from various stages of bovine embryo development. The microarray data analysis revealed that only a few genes were differentially expressed, showing the relative similarity between those two kinds of embryos. Nevertheless, the fact that we obtained a similar diversity of developmental stages with parthenotes suggests that synchronisation is more oocyte-specific than sperm entry-time related. We then analysed our data with Ingenuity pathway analysis. Networks of genes involved in blastocyst implantation but also previous stages of embryo development, like maternal-to-embryonic transition, were identified. This new information allows us to better understand the regulatory mechanisms of embryonic development associated with embryo status.
哺乳动物胚胎发育最初阶段的潜在过程仍未得到很好的描述。为了更好地确定受精后发生事件的动力学,最好使精子进入完全同步。由于同一组受精卵在6至12小时的时间间隔内发生受精,这导致胚胎发育激活时间的重大差异。牛孤雌生殖可能会实现更好的同步,如果是这样,将为研究发育能力提供更好的模型。在本研究中,牛卵母细胞要么进行孤雌激活,要么进行受精,并在体外培养7天。使用BlueChip对这两组早期和扩张期胚胎进行基因表达分析,BlueChip是我们实验室开发的定制2000-cDNA阵列,富含来自牛胚胎发育各个阶段的克隆。微阵列数据分析显示,只有少数基因存在差异表达,表明这两种胚胎之间具有相对相似性。然而,我们从孤雌胚胎获得了相似的发育阶段多样性这一事实表明,同步性更多地是卵母细胞特异性的,而与精子进入时间无关。然后,我们用Ingenuity通路分析来分析数据。识别出了参与囊胚着床以及胚胎发育先前阶段(如母源-胚胎转变)的基因网络。这些新信息使我们能够更好地理解与胚胎状态相关的胚胎发育调控机制。