Cloning, Characterization and Bioinformatics Analysis of the Sequences of miR-10a and miR-10b in Sheep (Hu sheep)

Abstract

Background: MicroRNAs (miRNAs) are active regulators of numerous biological and physiological processes and play an important role in the regulation of animal ovaries and other reproductive related organs. To understand the molecular mechanisms of miR-10 family, we investigated the molecular characteristics and the relative expression of sheep miR-10a and miR-10b (miR-10a/b) and conducted bioinformatics analysis.Methods: During the period 2018-2019 total of 20 samples including blood and tissues such as hypothalamus, pituitary and ovary were collected from the Hu sheep raised in the National Meat Sheep Experimental Station of China (Luoyang City, Henan Province, China). Blood was collected from jugular vein by vacuum and anticoagulation blood collection tube and stored in refrigerator at -20°C. The tissues were placed into the cryopreservation tube treated with Diethy pyrocarbonate (DEPC) water and stored in liquid nitrogen. All the samples were processed for isolation and confirmed with biochemical analysis and Polymerase chain reaction (PCR) and Real-time fluorescence quantitative PCR.Result: The target genes were predicted by three kinds of target gene predicting software. The function of target genes and their involved pathways were obtained by gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The relative expression of miR-10a/b in sheep ovary was extremely significantly higher than that in hypothalamus and pituitary gland (P less than 0.01). The relative expression of miR-10b in ovary or pituitary was extremely significantly higher than that of miR-10a (P less than 0.01) and the relative expression of miR-10b in hypothalamus was significantly higher than that of miR-10a (P less than 0.05). These results serve as a foundation for further study on the Sheep miR-10 family.