Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes
Open Access
- 31 December 2012
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 7 (12), e52592
- https://doi.org/10.1371/journal.pone.0052592
Abstract
A cultured C2C12 myotube contraction system was examined for application as a model for acute contraction-induced phenotypes of skeletal muscle. C2C12 myotubes seeded into 4-well rectangular plates were placed in a contraction system equipped with a carbon electrode at each end. The myotubes were stimulated with electric pulses of 50 V at 1 Hz for 3 ms at 997-ms intervals. Approximately 80% of the myotubes were observed to contract microscopically, and the contractions lasted for at least 3 h with electrical stimulation. Calcium ion (Ca2+) transient evoked by the electric pulses was detected fluorescently with Fluo-8. Phosphorylation of protein kinase B/Akt (Akt), 5′ AMP-activated protein kinase (AMPK), p38 mitogen-activated protein kinase (p38), and c-Jun NH2-terminal kinase (JNK)1/2, which are intracellular signaling proteins typically activated in exercised/contracted skeletal muscle, was observed in the electrically stimulated C2C12 myotubes. The contractions induced by the electric pulses increased glucose uptake and depleted glycogen in the C2C12 myotubes. C2C12 myotubes that differentiated after exogenous gene transfection by a lipofection or an electroporation method retained their normal contractile ability by electrical stimulation. These findings show that our C2C12 cell contraction system reproduces the muscle phenotypes that arise in vivo (exercise), in situ (hindlimb muscles in an anesthetized animal), and in vitro (dissected muscle tissues in incubation buffer) by acute muscle contraction, demonstrating that the system is applicable for the analysis of intracellular events evoked by acute muscle contraction.This publication has 21 references indexed in Scilit:
- Electrical Pulse Stimulation of Cultured Human Skeletal Muscle Cells as an In Vitro Model of ExercisePLOS ONE, 2012
- Designing of a Si-MEMS device with an integrated skeletal muscle cell-based bio-actuatorBiomedical Microdevices, 2010
- Measurement of Contractile Stress Generated by Cultured Rat Muscle on Silicon Cantilevers for Toxin Detection and Muscle Performance EnhancementPLOS ONE, 2010
- Micropatterning contractile C2C12 myotubes embedded in a fibrin gelBiotechnology & Bioengineering, 2009
- Contractile C2C12myotube model for studying exercise-inducible responses in skeletal muscleAmerican Journal of Physiology-Endocrinology and Metabolism, 2008
- Exercise-induced phospho-proteins in skeletal muscleInternational Journal of Obesity, 2008
- AMP-activated protein kinase and the regulation of glucose transportAmerican Journal of Physiology-Endocrinology and Metabolism, 2006
- Regulation of glucose transport by the AMP-activated protein kinaseProceedings of the Nutrition Society, 2004
- Contraction Regulation of Akt in Rat Skeletal MuscleOnline Journal of Public Health Informatics, 2002
- Exercise Stimulates c-Jun NH2Kinase Activity and c-Jun Transcriptional Activity in Human Skeletal MuscleBiochemical and Biophysical Research Communications, 1998