Abstract
BackgroundIntramuscular fat (IMF) content is an important factor in porcine meat quality. Previously, we showed that miR-34a was less abundant in liver tissue from pigs with higher backfat thickness, compared to pigs with lower backfat thickness. The purpose of this present study was to explore the role of miR-34a in adipogenesis.ResultBioinformatics analysis identified Acyl-CoA synthetase long chain family member 4 (ACSL4) as a putative target of miR-34a. Using a luciferase reporter assay, we verified that miR-34a binds the ACSL4 mRNA at the 3’UTR. To examine the role of the miR-34a-ACSL4 interaction in IMF deposition in the pig, mRNA and protein expression of the ACSL4 gene was measured in primary intramuscular preadipocytes transfected with miR-34a mimic and inhibitor. Our results showed that ACSL4 is expressed throughout the entire differentiation process in pig preadipocytes, similar to the lipogenesis-associated genes PPARγ and aP2. Transfection with miR-34a mimic reduced lipid droplet formation during adipogenesis, while miR-34a inhibitor increased lipid droplet accumulation. Transfection with miR-34a mimic also reduced the mRNA and protein expression of ACSL4 and lipogenesis genes, including PPARγ, aP2, and SREBP-1C, but increased the expression of steatolysis genes such as ATGL and Sirt1. In contrast, the miR-34a inhibitor had the opposite effect on gene expression. Further, knockdown of ACSL4 decreased lipid droplet accumulation.ConclusionsOur results support the hypothesis that miR-34a regulates intramuscular fat deposition in porcine adipocytes by targeting ACSL4.
Highlights
Intramuscular fat (IMF) content is an important factor in porcine meat quality
Two hundred ninety-eight genes overlapped with the targets (Supplementary Table S3), and were examined for potential biological roles using Gene Ontology (GO) term enrichment and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analyses
We investigated the mechanism by which miR-34a affects lipid metabolism in pigs
Summary
Intramuscular fat (IMF) content is an important factor in porcine meat quality. We showed that miR-34a was less abundant in liver tissue from pigs with higher backfat thickness, compared to pigs with lower backfat thickness. The purpose of this present study was to explore the role of miR-34a in adipogenesis. Intramuscular fat (IMF) content is a primary indicator of porcine meat quality [1]. An increase in IMF content can improve meat flavor [2]. Substantial efforts have been made to improve production efficiency and select for lean growth, both of which impact IMF negatively. Selection for enhanced IMF has become an important focus in pork production. MicroRNAs are small non-coding RNA molecules that regulate gene expression by targeting mRNA transcripts for cleavage or translational inhibition [10, 11]. miRNAs play important roles in various biological processes, including cell differentiation, proliferation and apoptosis [12], organ development [13], lipid metabolism [14], and
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