| Summary: | Intramuscular fat (IMF) is one of the crucial factors determining meat quality. IMF deposition depends on the hyperplasia and hypertrophy of intramuscular preadipocytes, in which genes and noncoding RNAs play an important regulatory role. According to previous transcriptome analysis, <i>ANXA6</i> and miR-24-3p were identified as involved in lipid metabolism in breast muscle. In this study, we further investigated their function in the proliferation and differentiation of chicken intramuscular preadipocytes. The results indicated that overexpression of <i>ANXA6</i> inhibited proliferation and promoted differentiation of intramuscular preadipocytes, while knockdown of <i>ANXA6</i> promoted cell proliferation and inhibited adipogenic differentiation. miR-24-3p was proved to directly bind to the 3′ untranslated region (3′UTR) of <i>ANXA6</i> by dual-luciferase reporter assay. The regulatory effect of miR-24-3p on the proliferation and differentiation of intramuscular preadipocytes was opposite to that of <i>ANXA6</i>. Besides, the overexpression vector of <i>ANXA6</i> eliminated the impact of miR-24-3p mimics on intramuscular preadipocytes. In brief, we revealed that miR-24-3p promoted proliferation but inhibited differentiation of intramuscular preadipocytes by blocking <i>ANXA6</i> expression, thus dominating IMF deposition in broilers. These findings may provide a novel target for improving chicken meat quality.
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