Abnormal DNA methylation of EBF1 regulates adipogenesis in chicken.

Background: DNA methylation influences gene expression and is involved in numerous biological processes, including fat production. It is involved in lipid generation in numerous animal species, including poultry. However, the effect of DNA methylation on adipogenesis in chickens remains unclear.

Results: A total of 12 100-day-old chickens were divided into high and low-fat groups based on their abdominal fat ratios. Subsequently, genome-wide bisulfite sequencing (WGBS) was performed on their abdominal fat, and 1877 differentially methylated region (DMR) genes were identified, among which SLC45A3, EBF1, PLA2G15, and ACAD9 were associated with lipid metabolism. Interestingly, EBF1 showed a lower level of DNA methylation and higher mRNA expression in the low-fat group, as determined by comprehensive RNA-seq analysis. Cellular verification showed that EBF1 expression was upregulated by 5-azacytidine (5-Aza) and downregulated by betaine. EBF1 facilitated the differentiation of immortalized chicken preadipocyte 1 (ICP-1) through the PPAR-γ pathway, thereby affecting chicken adipogenesis.

Conclusion: A combination of WGBS and RNA-seq analyses revealed 48 DMGs in the abdominal fat tissue of chickens. Notably, the DNA methylation status of EBF1 was inversely related to its mRNA expression. Mechanistically, DNA methylation regulates EBF1 expression, which in turn mediates the differentiation of ICP-1 through the PPARγ pathway. This study provides a theoretical framework for investigating the effects of DNA methylation on adipogenesis in chickens.