Abstract
Our previous studies showed that microRNA-15a (miR-15a) was closely related to intramuscular fat (IMF) deposition in chickens; however, its regulatory mechanism remains unclear. Here, we evaluated the expression characteristics of miR-15a and its relationship with the expression of acetyl-CoA acyltransferase 1 (ACAA1), acyl-CoA oxidase 1 (ACOX1) and sterol carrier protein 2 (SCP2) by qPCR analysis in Gushi chicken breast muscle at 6, 14, 22, and 30 weeks old, where we performed transfection tests of miR-15a mimics in intramuscular preadipocytes and verified the target gene of miR-15a in chicken fibroblasts (DF1). The miR-15a expression level at 30 weeks increased 13.5, 4.5, and 2.7-fold compared with the expression levels at 6, 14, and 22 weeks, respectively. After 6 days of induction, miR-15a over-expression significantly promoted intramuscular adipogenic differentiation and increased cholesterol and triglyceride accumulation in adipocytes. Meanwhile, 48 h after transfection with miR-15a mimics, the expression levels of ACAA1, ACOX1 and SCP2 genes decreased by 56.52%, 31.18% and 37.14% at the mRNA level in intramuscular preadipocytes. In addition, the co-transfection of miR-15a mimics and ACAA1, ACOX1 and SCP2 3′UTR (untranslated region) dual-luciferase vector significantly inhibited dual-luciferase activity in DF1 cells. Taken together, our data demonstrate that miR-15a can reduce fatty acid oxidation by targeting ACAA1, ACOX1, and SCP2, which subsequently indirectly promotes the differentiation of chicken intramuscular preadipocytes.
Highlights
The chicken (Gallus gallus) is an economically important agricultural animal
The expression profile of miR-15a was determined in Gushi chicken breast muscle tissues at 6, 14, 22, and 30 weeks old, and its expression level was significantly upregulated with the development of breast muscle (Figure 1)
The role of miR-15a was further determined in chicken intramuscular preadipocytes
Summary
The chicken (Gallus gallus) is an economically important agricultural animal. Intramuscular fat (IMF) is one of the major factors affecting the quality of poultry meat. Previous studies have found that miR-15a regulates cell size and proliferation by fine-tuning delta-like 1 homolog (DLK1) in 3T3-L1 preadipocytes [19] and promotes adipogenesis via repressing forkhead box protein O1 (FoxO1) in pigs [20] This suggested that miR-15a may affect IMF deposition by regulating various adipocyte events such as cell proliferation and differentiation in chickens. Based on the association analysis of miRNA and mRNA transcriptome profiles, we found that the dynamic expression trend of genes including acetyl-CoA acyltransferase 1 (ACAA1), sterol carrier protein 2 (SCP2) and acyl-CoA oxidase 1 (ACOX1) has a significant negative correlation with the dynamic changes of miR-15a in the breast muscle of Gushi chicken [24]. These results will contribute to a better understanding of the molecular mechanisms of IMF deposition in the chicken
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