Neutral sphingomyelinase-3 mediates TNF-stimulated oxidant activity in skeletal muscle

Jennifer S. Moylan, Jeffrey D. Smith, Erin M. Wolf Horrell, Julie B. McLean, Gergana M. Deevska, Mark R. Bonnell, Mariana N. Nikolova-Karakashian, Michael B. Reid

Research output: Contribution to journalArticlepeer-review

32 Scopus citations


Aims: Sphingolipid and oxidant signaling affect glucose uptake, atrophy, and force production of skeletal muscle similarly and both are stimulated by tumor necrosis factor (TNF), suggesting a connection between systems. Sphingolipid signaling is initiated by neutral sphingomyelinase (nSMase), a family of agonist-activated effector enzymes. Northern blot analyses suggest that nSMase3 may be a striated muscle-specific nSMase. The present study tested the hypothesis that nSMase3 protein is expressed in skeletal muscle and functions to regulate TNF-stimulated oxidant production. Results: We demonstrate constitutive nSMase activity in skeletal muscles of healthy mice and humans and in differentiated C2C12 myotubes. nSMase3 (Smpd4 gene) mRNA is highly expressed in muscle. An nSMase3 protein doublet (88 and 85. kD) is derived from alternative mRNA splicing of exon 11. The proteins partition differently. The full-length 88. kD isoform (nSMase3a) fractionates with membrane proteins that are resistant to detergent extraction; the 85. kD isoform lacking exon 11 (nSMase3b) is more readily extracted and fractionates with detergent soluble membrane proteins; neither variant is detected in the cytosol. By immunofluorescence microscopy, nSMase3 resides in both internal and sarcolemmal membranes. Finally, myotube nSMase activity and cytosolic oxidant activity are stimulated by TNF. Both if these responses are inhibited by nSMase3 knockdown. Innovation: These findings identify nSMase3 as an intermediate that links TNF receptor activation, sphingolipid signaling, and skeletal muscle oxidant production. Conclusion: Our data show that nSMase3 acts as a signaling nSMase in skeletal muscle that is essential for TNF-stimulated oxidant activity.

Original languageEnglish
Pages (from-to)910-920
Number of pages11
JournalRedox Biology
Issue number1
StatePublished - Oct 2014


  • Free radicals
  • NSMase3
  • Neutral sphingomyelinase-3
  • Oxidants
  • ROS
  • Reactive oxygen species
  • Skeletal muscle
  • Sphingolipid signaling

ASJC Scopus subject areas

  • Organic Chemistry


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