A potential regulatory network underlying distinct fate commitment of myogenic and adipogenic cells in skeletal muscle

Abstract

Mechanism controlling myo-adipogenic balance in skeletal muscle is of great significance for human skeletal muscle dysfunction and myopathies as well as livestock meat quality. In the present study, two cell subpopulations with particular potency of adipogenic or myogenic differentiation were isolated from neonatal porcine longissimus dorsi using the preplate method to detect mechanisms underlying distinct fate commitment of myogenic and adipogenic cells in skeletal muscle. Both cells share a common surface expression profile of CD29+CD31−CD34−CD90+CD105+, verifying their mesenchymal origin. A total of 448 differentially expressed genes (DEGs) (FDR < 0.05 and |log2 FC| ≥ 1) between two distinct cells were identified via RNA-seq, including 358 up-regulated and 90 down-regulated genes in myogenic cells compared with adipogenic cells. The results of functional annotation and enrichment showed that 42 DEGs were implicated in cell differentiation, among them PDGFRα, ITGA3, ITGB6, MLCK and MLC acted as hubs between environment information processing and cellular process, indicating that the interaction of the two categories exerts an important role in distinct fate commitment of myogenic and adipogenic cells. Particularly, we are first to show that up-regulation of intracellular Ca2+-MLCK and Rho-DMPK, and subsequently elevated MLC, may contribute to the distinct commitment of myogenic and adipogenic lineages via mediating cytoskeleton dynamics.

Association Analysis of Single-Cell RNA Sequencing and Proteomics Reveals a Vital Role of Ca2+ Signaling in the Determination of Skeletal Muscle Development Potential

“…Ca 2+ , serving as the second messenger, regulates numerous cellular processes at the center of cell signaling [54] and has been shown to participate in the regulation of myogenic differentiation [35,55]. In this study, Ca 2+ concentration in both cytoplasmic and endoplasmic reticulum was lower in Myo-lineage cells of lean-type pigs relative to obese-type pigs.…”

“…Besides, the global proteomic profiles of Myo-lineage cells revealed that major differences between obese-and lean-type pigs belong to categories that include muscle development, ion homeostasis, apoptosis, and the MAPK signaling pathway. In a previous study, we showed that the MAPK signaling pathway participates in the mediation of myogenic differentiation [34,35]. Thereby, in the present study, we mainly focused on the proteomic landscape among cell proliferation and differentiation, cellular ion homeostasis, and muscle development.…”

“…PALLD can enhance the migration activity of cells by changing the state of the actin cytoskeleton and promote cell differentiation and skeletal muscle maturation [41]. FN1 regulated cell differentiation fate by affecting cytoskeletal status [35]. Both of PALLD and FN1 were upregulated in the Myo-lineage cells of lean-type pigs, while GPCPD1, catalyzing the phosphorylation of choline and thus promoting lipid metabolism [42], was downregulated.…”

Dynamic membrane proteome of adipogenic and myogenic precursors in skeletal muscle highlights EPHA2 may promote myogenic differentiation through ERK signaling

“…For pigs, cell surface marker of muscle-derived cells has not been established because of the limited knowledge of cell surface markers and available commercialized antibodies. Therefore, we had successfully established a preplate technique, a cell surface marker–-independent method, to isolate adipogenic and myogenic precursors from skeletal muscle of pigs ( 24 , 25 ) by adapting the previous protocol ( 26 ) with some modifications. In this study, we proved again the purity of adipogenic and myogenic precursors by their distinct differentiation capacity upon adipogenic and myogenic induction (as shown in Fig.…”

“…Adipogenic and myogenic precursors were isolated from the longissimus dorsi muscle of 3-d-old newborn Yorkshire pigs according to the protocol described in our previous study ( 24 , 25 ) by adapting the previous protocol ( 26 ) with some modifications. Briefly, skeletal muscles were minced and digested in 0.17% protease (P8811; MilliporeSigma, Burlington, MA, USA) and 0.15% collagenase-type XI (C9407; MilliporeSigma) solution for 1 h, respectively.…”