Differential chromatin accessibility between pre- and post-natal stages highlights putative causal regulatory variants in pig skeletal muscle

Abstract

Deciphering how chromatin dynamics shape the effects of regulatory variants on complex traits across development remains largely unexplored in farmed animals. Focusing on porcine skeletal muscle, we integrated chromatin accessibility (ATAC-seq), molecular QTL (molQTL), and GWAS data to identify putative stage-specific regulatory variants influencing agronomically important traits. We aggregated and analyzed 202 ATAC-seq libraries spanning prenatal (fetal) and postnatal (piglet) stages. We intersected consensus and differential peaks with cis-molQTLs from the Porcine Genotype-by-Tissue-by-Expression Atlas (PigGTEx) followed by functional enrichment analysis, promoter-focused functional annotation, integration with gene-trait pairs, and intersection with eGenes (i.e., genes whose expression is regulated by at least one cis-eQTL) to link variants previously associated with QTLs for production traits. We identified 132,275 differentially accessible peaks (DAPs) distinguishing fetal and piglet muscle. Fetal DAPs were enriched in promoter and intergenic regions, whereas piglet DAPs were enriched in intronic regions, indicating a shift from transcriptional priming to postnatal regulatory refinement. Overlaps between accessible regions and molQTLs were highly significant (p < 0.001), with the strongest eQTLs predominating in the fetal stage (14 fetal vs. 4 piglet). Gene Ontology analysis of promoter-accessible eGenes revealed enrichment for RNA metabolism and chromatin organization in fetuses, and for muscle contraction and lipid metabolism in piglets. The intersection with colocalized complex traits loci identified 107 fetal- and 30 piglet-stage regulatory elements, with only three eGenes shared between stages. These findings provide insights into developmental chromatin dynamics in skeletal muscle and an effective framework for prioritising putative regulatory variants affecting pig traits at prenatal stages.