Chair of Animal Breeding and Biotechnology, Institute of Veterinary Medicine and Animal Sciences, Estonian University of Life Sciences, Kreutzwaldi 62, 51014 Tartu, Estonia.
Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, 50411 Tartu, Estonia.
Genes (Basel). 2023 Feb 24;14(3):569. doi: 10.3390/genes14030569.
Recent advances in preimplantation embryo diagnostics enable a wide range of applications using single cell biopsy and molecular-based selection techniques without compromising embryo production. This study was conducted to develop a single cell embryo biopsy technique and gene expression analysis method with a very low input volume to ensure normal embryo development and to see if there are differences in gene expression profiles between day-5 biopsied bovine embryos that developed into blastocysts and embryos arrested at morula stage. Out of the 65 biopsied morulae, 32 developed to blastocysts (49.2%). Out of the 13,580 successfully annotated genes, 1204 showed a difference in mRNA expression level. Out of these, 155 genes were expressed in embryos developing to blastocysts. The pathway enrichment analysis revealed significant enrichment in "organelle biogenesis and maintenance", "mRNA splicing" and "mitochondrial translation" pathways. These findings suggest principal differences in gene expression patterns and functional networks of embryos able to reach the blastocyst stage compared to embryos arrested in development. Our preliminary data suggest that single blastomere biopsy and selected gene expression profiles at morula stage could offer additional possibilities for early preimplantation embryo selection before transfer.
胚胎植入前诊断的最新进展使得利用单细胞活检和基于分子的选择技术进行广泛的应用成为可能,而不会影响胚胎的产生。本研究旨在开发一种单细胞胚胎活检技术和基因表达分析方法,其输入量非常低,以确保胚胎的正常发育,并观察在第 5 天进行活检的牛胚胎中,是否存在发育为囊胚和停滞在桑葚胚阶段的胚胎之间基因表达谱的差异。在 65 个活检桑葚胚中,有 32 个发育为囊胚(49.2%)。在成功注释的 13580 个基因中,有 1204 个显示出 mRNA 表达水平的差异。其中,155 个基因在发育为囊胚的胚胎中表达。通路富集分析显示,“细胞器生物发生和维持”、“mRNA 剪接”和“线粒体翻译”通路显著富集。这些发现表明,与发育停滞的胚胎相比,能够到达囊胚阶段的胚胎在基因表达模式和功能网络上存在主要差异。我们的初步数据表明,在转移前的早期胚胎选择中,单个卵裂球活检和桑葚胚阶段的特定基因表达谱可能提供额外的可能性。