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Updated: Mar 10, 2026
Modeling Myotonic Dystrophy 1 in C2C12 Myoblast Cells
Published on: July 29, 2016
N-methyl-D-aspartate (NMDA) impairs myogenesis in C2C12 cells
Q-SChick Auh1, Kyung-Ran Park2, Myeong-Ok Lee3
1Department of Oral Medicine, School of Dentistry, Kyung Hee University, Seoul, Korea.
Introduction:
N-methyl-d-aspartate (NMDA) is expressed in sensory neurons and plays important roles in peripheral pain mechanisms. The aim of this study was to examine the effects and molecular mechanisms of NMDA on C2C12 myoblast proliferation and differentiation.
Methods:
Cytotoxicity and differentiation were examined by the MTT assay, reverse transcription-polymerase chain reaction, and immunofluorescence.
Results:
NMDA had no cytotoxicity (10-500 μM) and inhibited myoblastic differentiation of C2C12 cells, as assessed by F-actin immunofluorescence and levels of mRNAs encoding myogenic markers such as myogenin and myosin heavy-chain 2. It inhibited phosphorylation of mammalian target of rapamycin (mTOR) by inactivating mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38. It induced reactive oxygen species production. Furthermore, NMDA-suppressed expression of F-actin was reversed by adding the antioxidant N-acetylcysteine.
Conclusions:
Collectively, these results indicate that NMDA impairs myogenesis or myogenic differentiation in C2C12 cells through the mTOR/MAPK signaling pathways and may lead to skeletal muscle degeneration. Muscle Nerve 56: 510-518, 2017.
Insights
N-methyl-d-aspartate (NMDA) inhibits C2C12 myoblast differentiation by affecting mTOR/MAPK signaling and increasing oxidative stress. Antioxidants may counteract these effects, suggesting NMDA
Area of Science:
- Cell Biology
- Neuroscience
- Muscle Physiology
Background:
- N-methyl-d-aspartate (NMDA) is involved in pain mechanisms.
- The role of NMDA in muscle cell differentiation is not fully understood.
Purpose of the Study:
- To investigate the effects of NMDA on C2C12 myoblast proliferation and differentiation.
- To elucidate the molecular mechanisms underlying NMDA's actions on myogenesis.
Main Methods:
- C2C12 myoblasts were treated with NMDA.
- Cytotoxicity was assessed using MTT assay.
- Differentiation was evaluated via immunofluorescence and RT-PCR for myogenic markers.
Main Results:
- NMDA (10-500 μM) showed no cytotoxicity.
- NMDA inhibited C2C12 myoblast differentiation, reducing F-actin and myogenic marker expression.
- NMDA inactivated mTOR/MAPK signaling pathways and induced reactive oxygen species (ROS).
- N-acetylcysteine reversed NMDA-induced F-actin suppression.
Conclusions:
- NMDA impairs myogenesis and myogenic differentiation in C2C12 cells.
- The mTOR/MAPK signaling pathway and oxidative stress are implicated in NMDA's effects.
- NMDA may contribute to skeletal muscle degeneration.
