miR-17-5p Regulates Differential Expression of NCOA3 in Pig Intramuscular and Subcutaneous Adipose Tissue

Abstract

Fat distribution affects economic value in pork production. Intramuscular adipose tissue (IMAT) improves meat quality, whereas subcutaneous adipose tissue (SCAT) is usually regarded as waste. In the present study, we analyzed IMAT/SCAT (I/S) ratios in each pig. Individuals selected from a population of 1200 Suhuai pigs were divided into two cohorts; those with high I/S ratios and those with low I/S ratios, and correlations between nuclear Receptor Co-activator 3 (NCOA3), a critical gene involved in regulating fat accumulation, and fat distribution were investigated. The ratio of IMAT NCOA3 to SCAT NCOA3 expression levels (NCOA3_I/NCOA3_S) was higher in the high I/S group compared with the low I/S group. The NCOA3 expression level in fat tissue was positively correlated with fat deposition. miR-17-5p was identified as a putative regulator of NCOA3 based on bioinformatics prediction analysis followed by gene expression analysis. The miR-17-5p_I/miR-17-5p_S ratio was negatively correlated with the NCOA3_I/NCOA3_S ratio. The predicted relationship between miR-17-5p and NCOA3 was further verified by dual luciferase activity assays, qPCR, and western blots. Overexpression of miR-17-5p in intramuscular preadipocytes inhibited NCOA3 expression and reduced preadipocyte differentiation. FABP4 and PPARG expression were also significantly decreased, as was triglyceride content. Meanwhile, knockdown of miR-17-5p significantly increased NCOA3 expression and promoted intramuscular preadipocyte differentiation. Based on these results, we propose that differential expression of NCOA3 in pig intramuscular and subcutaneous adipose tissue is regulated by miR-17-5p.

Keywords: Fat differentiation; IMAT; NCOA3; SCAT; miR-17-5p.