Abstract
Intramuscular fat (i.m.) is an adipose tissue that is deposited between muscle bundles. An important type of post-transcriptional regulatory factor, miRNAs, has been observed as an important regulator that can regulate gene expression and cell differentiation through specific binding with target genes, which is the pivotal way determining intramuscular fat deposition. Thus, this study intends to use RT-PCR, cell culture, liposome transfection, real-time fluorescent quantitative PCR (qPCR), dual luciferase reporter systems, and other biological methods clarifying the possible mechanisms on goat intramuscular preadipocyte differentiation that is regulated by miR-214-5p. Ultimately, our results showed that the expression level of miR-214-5p peaked at 48 h after the goat intramuscular preadipocytes were induced for adipogenesis. Furthermore, after inhibition of the expression of miR-214-5p, the accumulation of lipid droplets and adipocyte differentiation in goat intramuscular adipocytes were promoted by the way of up-regulation of the expression level of lipoprotein lipase (LPL) (p < 0.05) and peroxisome proliferator-activated receptor gamma (PPARγ) (p < 0.01) but inhibited the expression of hormone-sensitive lipase (HSL) (p < 0.01). Subsequently, our study confirmed that Krüppel-like factor 12 (KLF12) was the target gene of miR-214-5p. Inhibition of the expression of KLF12 promoted adipocyte differentiation and lipid accumulation by upregulation of the expression of LPL and CCAAT/enhancer binding protein (C/EBPα) (p < 0.01). Overall, these results indicated that miR-214-5p and its target gene KLF12 were negative regulators in progression of goat preadipocyte differentiation. Our research results provided an experimental basis for finally revealing the mechanism of miR-214-5p in adipocytes.
Highlights
Intramuscular fat (i.m.) content is an extremely important indicator that affects the tenderness, flavor, and juiciness of goat meat; the intramuscular fat deposition mainly depends on the differentiation of intramuscular preadipocytes and the accumulation of triglycerides
The qRT-polymerase chain reaction (PCR) technique was used for detecting the expression level of miR-214-5p after induced adipogenesis for 0–96 h (Figure 1B), and our results showed that the expression level of miR-214-5p peaked at 48 h, which was significantly higher than that at 0 h (p < 0.01)
Our results showed that compared with NC, the expression levels of lipoprotein lipase (LPL), ACC, and peroxisome proliferator-activated receptor gamma (PPARc) were significantly upregulated after inhibiting miR-214-5p, while the expression of hormone-sensitive lipase (HSL) was significantly downregulated
Summary
Intramuscular fat (i.m.) content is an extremely important indicator that affects the tenderness, flavor, and juiciness of goat meat; the intramuscular fat deposition mainly depends on the differentiation of intramuscular preadipocytes and the accumulation of triglycerides. With the general application of genome sequencing, researchers found that the complexity of biology is the difference in the proportion of non-protein-coding genomes, and most of the long or small nonmiR-214-5p Regulating Preadipocytes Differentiation coding RNAs coordinated protein expression at transcription or translation levels (Taft et al, 2007; Vienberg et al, 2017). Each miRNA can regulate hundreds of target genes’ expression by the way of induced translational inhibition or degradation of transcription products of the target gene via binding to the complementary sites of the 3′-untranslated region (3′UTR) (Hammond, 2015; Colamatteo et al, 2019; Roberts, 2015; Hammond, 2015; Roberts, 2015; Colamatteo et al, 2019). Elucidating the key genes and molecular regulatory networks during differentiation of adipocytes is essential for understanding the physiological process of adipogenesis
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