Hitit Mustafa, Kose Mehmet, Kaya Mehmet Salih, Kırbas Mesut, Dursun Sukru, Alak Ilyas, Atli Mehmet Osman
Department of Genetics, Faculty of Veterinary Medicine, Kastamonu University, Kastamonu, Turkey.
Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Dicle University, Diyarbakir, Turkey.
Front Genet. 2022 Aug 17;13:929477. doi: 10.3389/fgene.2022.929477. eCollection 2022.
MicroRNA (miRNA) plays an important role in the control of gene expression and is implied in many biological functions, including embryo implantation and development. The aim was to assess plasma miRNA profiles during the peri-implantation and ascertain potential candidate miRNA markers for early pregnancy diagnosis in ovine plasma. The plasma samples were obtained from a total of 24 ewes on days 12 (pre-implantation; P12, = 4), 16 (implantation; P16, = 4) and 22 (post-implantation; P22, = 4) after mating, and on their corresponding days of 12 (Pre-C; C12, = 4), 16 (Imp-C; C16, = 4) and 22 (Post-C; C22, = 4) of the estrous cycle. The miRNA profiles in plasma were assessed by microarray technology. We detected the presence of 60 ovine-specific miRNAs in plasma samples. Of these miRNAs, 22 demonstrated a differential expression pattern, especially between the estrous cycle and early pregnancy, and targeted 521 genes. Two miRNAs (oar-miR-218a and oar-miR-1185-3p) were confirmed using RT-qPCR in the ovine plasma samples. Protein-protein interaction (PPI) network of target genes established six functional modules, of which modules 1 and 3 were enriched in the common GO terms, such as inflammatory response, defense response, and regulation of immune response. In contrast, module 2 was enriched in the developmental process involved in reproduction, embryo development, embryonic morphogenesis, and regulation of the developmental process. The results indicate that miRNAs profiles of plasma seemed to be modulated during the peri-implantation stage of pregnancy in ewes. Circulating miRNAs could be promising candidates for diagnosis in early ovine pregnancy.
微小RNA（miRNA）在基因表达调控中发挥着重要作用，并参与许多生物学功能，包括胚胎着床和发育。本研究旨在评估绵羊着床前后血浆中的miRNA谱，并确定用于早期妊娠诊断的潜在候选miRNA标志物。在交配后第12天（着床前；P12，n = 4）、第16天（着床期；P16，n = 4）和第22天（着床后；P22，n = 4），以及发情周期相应的第12天（对照12；C12，n = 4）、第16天（对照16；C16，n = 4）和第22天（对照22；C22，n = 4），从总共24只母羊采集血浆样本。采用微阵列技术评估血浆中的miRNA谱。我们在血浆样本中检测到60种绵羊特异性miRNA。其中22种表现出差异表达模式，尤其是在发情周期和早期妊娠之间，并且靶向521个基因。使用RT-qPCR在绵羊血浆样本中验证了两种miRNA（oar-miR-218a和oar-miR-1185-3p）。靶基因的蛋白质-蛋白质相互作用（PPI）网络建立了六个功能模块，其中模块1和3富含常见的基因本体（GO）术语，如炎症反应、防御反应和免疫反应调节。相比之下，模块2富含与生殖、胚胎发育、胚胎形态发生和发育过程调节相关的发育过程。结果表明，母羊妊娠着床前后阶段血浆中的miRNA谱似乎受到调控。循环miRNA可能是早期绵羊妊娠诊断的有前景的候选标志物。
