Primary bovine skeletal muscle cells enters apoptosis rapidly via the intrinsic pathway when available oxygen is removed

PLoS One. 2017 Aug 8;12(8):e0182928. doi: 10.1371/journal.pone.0182928. eCollection 2017.

Abstract

Muscle cells undergo changes post-mortem during the process of converting muscle into meat, and this complex process is far from revealed. Recent reports have suggested programmed cell death (apoptosis) to be important in the very early period of converting muscle into meat. The dynamic balance that occurs between anti-apoptotic members, such as Bcl-2, and pro-apoptotic members (Bid, Bim) helps determine whether the cell initiates apoptosis. In this study, we used primary bovine skeletal muscle cells, cultured in monolayers in vitro, to investigate if apoptosis is induced when oxygen is removed from the growth medium. Primary bovine muscle cells were differentiated to form myotubes, and anoxia was induced for 6h. The anoxic conditions significantly increased (P<0.05) the relative gene expression of anti- and pro-apoptotic markers (Aif, Bcl-2, Bid and Bim), and the PARK7 (P<0.05) and Grp75 (Hsp70) protein expressions were transiently increased. The anoxic conditions also led to a loss of mitochondrial membrane potential, which is an early apoptotic event, as well as cytochrome c release from the mitochondria. Finally, reorganization and degradation of cytoskeletal filaments occurred. These results suggest that muscle cells enters apoptosis via the intrinsic pathway rapidly when available oxygen in the muscle diminishes post-mortem.

MeSH terms

  • Animals
  • Apoptosis / physiology*
  • Blotting, Western
  • Cattle / metabolism*
  • Cell Hypoxia / physiology*
  • Cell Survival
  • Cells, Cultured
  • Cytochromes c / metabolism
  • Fluorescent Antibody Technique
  • Gene Expression
  • Meat Products
  • Membrane Potential, Mitochondrial / physiology
  • Microscopy, Fluorescence
  • Muscle Fibers, Skeletal / metabolism*
  • Muscle Fibers, Skeletal / pathology
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction
  • Tissue Culture Techniques

Substances

  • Cytochromes c

Grants and funding

This work was supported by grants from the Fund for Research Levy on Agricultural Products in Norway and the Norwegian Research Council (203697/O99). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.