Cessation of biomechanical stretch model of C2C12 cells models myocyte atrophy and anaplerotic changes in metabolism using non-targeted metabolomics analysis

Amro Ilaiwy, Megan T. Quintana, James R. Bain, Michael J. Muehlbauer, David I. Brown, William E. Stansfield, Monte Willis

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Studies of skeletal muscle disuse, either in patients on bed rest or experimentally in animals (immobilization), have demonstrated that decreased protein synthesis is common, with transient parallel increases in protein degradation. Muscle disuse atrophy involves a process of transition from slow to fast myosin fiber types. A shift toward glycolysis, decreased capacity for fat oxidation, and substrate accumulation in atrophied muscles have been reported, as has accommodation of the liver with an increased gluconeogenic capacity. Recent studies have modeled skeletal muscle disuse by using cyclic stretch of differentiated myotubes (C2C12), which mimics the loading pattern of mature skeletal muscle, followed by cessation of stretch. We utilized this model to determine the metabolic changes using non-targeted metabolomics analysis of the media. We identified increases in amino acids resulting from muscle atrophy-induced protein degradation (largely sarcomere) that occurs with muscle atrophy that are involved in feeding the Kreb's cycle through anaplerosis. Specifically, we identified increased alanine/proline metabolism (significantly elevated proline, alanine, glutamine, and asparagine) and increased α-ketoglutaric acid, the proposed Kreb's cycle intermediate being fed by the alanine/proline metabolic anaplerotic mechanism. Additionally, several unique pathways not clearly delineated in previous studies of muscle unloading were seen, including: (1) elevated keto-acids derived from branched chain amino acids (i.e. 2-ketoleucine and 2-keovaline), which feed into a metabolic pathway supplying acetyl-CoA and 2-hydroxybutyrate (also significantly increased); and (2) elevated guanine, an intermediate of purine metabolism, was seen at 12 h unloading. Given the interest in targeting different aspects of the ubiquitin proteasome system to inhibit protein degradation, this C2C12 system may allow the identification of direct and indirect alterations in metabolism due to anaplerosis or through other yet to be identified mechanisms using a non-targeted metabolomics approach.

Original languageEnglish (US)
Pages (from-to)80-92
Number of pages13
JournalInternational Journal of Biochemistry and Cell Biology
Volume79
DOIs
StatePublished - Oct 1 2016
Externally publishedYes

Fingerprint

Metabolomics
Muscular Atrophy
Metabolism
Muscle Cells
Proteolysis
Atrophy
Muscle
Skeletal Muscle
Proline
Alanine
Atrophic Muscular Disorders
Hydroxybutyrates
Keto Acids
Ketoglutaric Acids
Muscles
Branched Chain Amino Acids
Sarcomeres
Bed Rest
Acetyl Coenzyme A
Skeletal Muscle Fibers

Keywords

  • Anaplerosis
  • Atrophy
  • Biaxial stretch
  • C2C12
  • Hypertrophy regression
  • Metabolomics

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Cessation of biomechanical stretch model of C2C12 cells models myocyte atrophy and anaplerotic changes in metabolism using non-targeted metabolomics analysis. / Ilaiwy, Amro; Quintana, Megan T.; Bain, James R.; Muehlbauer, Michael J.; Brown, David I.; Stansfield, William E.; Willis, Monte.

In: International Journal of Biochemistry and Cell Biology, Vol. 79, 01.10.2016, p. 80-92.

Research output: Contribution to journalArticle

Ilaiwy, Amro ; Quintana, Megan T. ; Bain, James R. ; Muehlbauer, Michael J. ; Brown, David I. ; Stansfield, William E. ; Willis, Monte. / Cessation of biomechanical stretch model of C2C12 cells models myocyte atrophy and anaplerotic changes in metabolism using non-targeted metabolomics analysis. In: International Journal of Biochemistry and Cell Biology. 2016 ; Vol. 79. pp. 80-92.
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AU - Quintana, Megan T.

AU - Bain, James R.

AU - Muehlbauer, Michael J.

AU - Brown, David I.

AU - Stansfield, William E.

AU - Willis, Monte

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