MicroRNA profiles of fibroblasts derived from in vivo fertilized and fat-1 transgenic cattle

Fat-1 transgenic cattle have high levels of ω-3 fatty acids, which regulate several genes in fatty acid metabolism. In the current study, fibroblasts derived from in vivo fertilized (Ferti) and fat-1 transgenic (TG) Luxi cattle (Bos taurus), a local breed in China, were cultured and their miRNA expression was characterized. Expression of 352 known miRNAs differed in cells from Ferti and TG cattle: 83 miRNAs were found to be specifically expressed in cells from Ferti cattle while 23 miRNAs were found to be specifically expressed in cells from TG cattle. Novel differences in miRNA expression were also found in cells from Ferti and TG cattle. The identity of seven differentially expressed miRNAs was verified using quantitative real-time PCR, and target genes were identified computationally. GO and KEGG analysis revealed that these miRNAs were involved in seven major biological pathways, including metabolism, MAPK signaling, calcium signaling, purine metabolism, ubiquitin mediated proteolysis, pyrimidine metabolism, and the cell cycle. Overexpression of one of these miRNAs, miR-21-5p, was found to suppress expression of fibroblast growth factor 10 (FGF10) and adipose triglyceride lipase (ATGL) in fibroblasts from TG cattle and 3T3-L1 pre-adipocytes. Conversely, knockdown of miR-21-5p stimulated expression. Together, these results suggest that miRNAs potentially play a role in expression of lipogenic and lipolytic genes as well as in synthesis of ω-3 fatty acids facilitated by fat-1.