International Journal of Molecular Sciences · 2016 · 56 citations · 45 references
Life SciencesAnimal SciencesMolecular BiologyGene ExpressionLactationMetabolic RegulationFatty Acid MetabolismLipid MetabolismMammary Epithelial Cell BiologyMicrorna BiologyMicrorna-mediated RegulationMilk LipidsFatty Acid Metabolism During LactationLuciferase Reporter AssaysLuciferase ReportersMammary GlandsMicrorna In MetabolismMicrorna ScreeningMilk Fat BiosynthesisWestern Blot Analysis
MicroRNA (miRNA) regulates the expression of genes and influences a series of biological processes, including fatty acid metabolism. We screened the expression of miRNA in goat mammary glands during peak-lactation and non-lactating (“dry”) periods, and performed an in vitro study with goat mammary epithelial cells (GMEC) prior to sequencing analysis. Results illustrated that miR-30e-5p and miR-15a were highly expressed. Utilizing a luciferase reporter assay and Western blot, low-density lipoprotein receptor-related protein 6 (LRP6) and Yes associated protein 1 (YAP1) genes were demonstrated to be a target of miR-30e-5p and miR-15a in GMEC. Moreover, we demonstrated that the overexpression of miR-30e-5p and miR-15a in GMEC promoted fat metabolism while their knockdown impaired fat metabolism. These findings extend the discovery of a key role of miR-30e-5p and miR-15a in mediating adipocyte differentiation by suggesting a role in promoting milk fat synthesis. In conclusion, our findings indicate that miR-30e-5p, together with miR-15a, represses expression of LRP6 and promotes fat metabolism in GMEC. The data expanded our knowledge on the function of miRNAs in milk fat metabolism and synthesis in ruminant mammary cells.
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