Rio Paul Del, DiMarco Sierra, Madan Pavneesh
Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, 50 Stone Road East, Guelph, ON N1G 2W1, Canada.
Animals (Basel). 2024 Aug 13;14(16):2331. doi: 10.3390/ani14162331.
In bovine embryos, the microRNA (miRNA) expression has been profiled at each stage of early development in vitro. The miRNAomic analysis of spent media has the potential to reveal characteristics of embryo health; however, applications are limited without categorizing miRNA profiles by embryo quality. Time-lapse imaging has shown the timing of embryo development in vitro may be indicative of their developmental potential. The study aimed to characterize miRNAs in the spent media of bovine embryos with different growth rates during the pre-implantation phase. Bovine cumulus-oocyte complexes were aspirated from ovaries, fertilized, and cultured to blastocyst stage of development. At the 2-cell, 8-cell, and blastocyst stage, each microdrop of 30 presumptive zygotes were classified as slow- or fast-growing based on the percentage of embryos that had reached the desired morphological stage. A comparative analysis was performed on the spent media of slow- and fast-growing embryos using the results of a GeneChip miRNA 4.0 array hybridization. In total, 34 differentially expressed miRNAs were identified between the comparison groups: 14 miRNAs were found in the 2-cell samples, 7 in the 8-cell samples, and 12 in the blastocyst samples. The results demonstrate distinct miRNAs populations can be identified between slow- and fast-growing embryos, highlighting the novel biomarkers of developmental potential at each stage of pre-implantation development.
在牛胚胎中，已对体外早期发育的每个阶段的微小RNA（miRNA）表达进行了分析。对废弃培养液的miRNA组学分析有可能揭示胚胎健康的特征；然而，在未按胚胎质量对miRNA谱进行分类的情况下，其应用受到限制。延时成像显示，体外胚胎发育的时间可能表明其发育潜力。该研究旨在表征着床前阶段不同生长速率的牛胚胎废弃培养液中的miRNA。从卵巢中吸出牛卵丘-卵母细胞复合体，进行受精，并培养至囊胚发育阶段。在2细胞、8细胞和囊胚阶段，根据达到所需形态阶段的胚胎百分比，将30个推定受精卵的每个微滴分类为生长缓慢或快速生长。使用基因芯片miRNA 4.0阵列杂交的结果，对生长缓慢和快速生长的胚胎的废弃培养液进行了比较分析。在比较组之间总共鉴定出34个差异表达的miRNA：在2细胞样本中发现14个miRNA，在8细胞样本中发现7个，在囊胚样本中发现12个。结果表明，在生长缓慢和快速生长的胚胎之间可以鉴定出不同的miRNA群体，突出了着床前发育各阶段发育潜力的新型生物标志物。
