Single-cell transcriptional profiling of porcine muscle satellite cells and myoblasts during myogenesis

Abstract

Here, we unravel the transcriptional mechanisms that govern porcine muscle myogenesis, a crucial aspect of pork production. Through single cell RNA-sequencing of longissimus dorsi (LD) muscle from 3-day-old piglets, we delineated 5 major cell types from 14,002 cells—muscle cells, fibro-adipogenic progenitors, mural cells, endothelial cells, and immune cells. Within the muscle cell clusters, we specifically identified muscle satellite cells and myoblasts, with 5,388 differentially expressed genes distinguishing the cell populations. Furthermore, our investigation led to the identification of novel cell surface markers for porcine muscle satellite cells, namely ITGA7, SDC2, and SDC4. In addition, we found 55 transcription factors, including AHCTF1, CEBPD, and MAX, which contribute to the regulation of muscle development. Through predictive receptor–ligand analysis, we uncovered 16 secretory factors with associated cellular communication channels influencing porcine skeletal muscle development. Notably, we identified the secretory factor ANGPTL4 as an inhibitor of porcine muscle satellite cell proliferation and differentiation, and confirmed its function through the in vitro treatment of cell lines. We propose that ANGTPL4 is a potential target for the treatment of myopathy. Taken together, our findings contribute to the identification and characterization of myogenic genes and regulatory networks, which enhances our understanding of porcine muscle satellite cell myogenesis and offers potential avenues for optimizing pork production.