Single-Cell RNA Sequencing Reveals Dynamic Intercellular Communication Networks During Chicken Skeletal Muscle Development

Intercellular communication is crucial for the coordination of skeletal muscle development. However, the intricate signaling networks that regulate chicken myogenesis are not yet fully elucidated. In this study, we utilized CellChat analysis on single-cell and single-nucleus RNA sequencing data to systematically delineate cell–cell communication patterns across five critical developmental stages of chicken skeletal muscle: embryonic day 4 (E4), day 6 (E6), day 12 (E12), day 18 (E18), and post-hatch day 30 (P30). Our findings indicate that communication architectures are highly stage-specific, with mesenchymal cells acting as the predominant signaling hub during the early embryonic stages (E4–E6), whereas fibro-adipogenic progenitors become the principal communicators during mid-to-late embryogenesis (E12–E18). At E4, the communication network was relatively simple, comprising 51 ligand–receptor pairs primarily involving the neural cell adhesion molecule, slit guidance ligand, and midkine (MK) signaling pathways between myogenic progenitors and mesenchymal cells. By E6, the network had expanded significantly, encompassing 6237 ligand–receptor pairs across 51 signaling pathways, which coincided with the emergence of multiple myogenic lineages. Peak communication complexity was observed at E12, characterized by 11,675 ligand–receptor pairs and 61 signaling pathways, reflecting the secondary wave of myogenesis. Comparative analysis across developmental stages revealed key signaling transitions: the pleiotrophin and MK pathways were predominantly active during the early phase of myogenic commitment (E4–E6), whereas the collagen, laminin, and adhesion G protein-coupled receptor L pathways were more prominent during the secondary myogenesis phase (E6–E12). Notably, a significant shift in communication patterns was observed from E12 to E18, marked by a reduction in developmental pathway signaling and an increase in immune-related communications. By P30, the communication network had stabilized into a homeostatic state, centered on interactions among myofibers, stromal cells, and the vascular system. This comprehensive atlas of intercellular communication offers novel insights into the signaling dynamics underpinning chicken skeletal muscle development.