Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes
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 st...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Online |
| Language: | English |
| Published: |
Public Library of Science
2012
|
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3534077/ |
| id |
pubmed-3534077 |
|---|---|
| recordtype |
oai_dc |
| spelling |
pubmed-35340772013-01-08 Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes Manabe, Yasuko Miyatake, Shouta Takagi, Mayumi Nakamura, Mio Okeda, Ai Nakano, Taemi Hirshman, Michael F. Goodyear, Laurie J. Fujii, Nobuharu L. Research Article 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. Public Library of Science 2012-12-31 /pmc/articles/PMC3534077/ /pubmed/23300713 http://dx.doi.org/10.1371/journal.pone.0052592 Text en © 2012 Manabe et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Manabe, Yasuko Miyatake, Shouta Takagi, Mayumi Nakamura, Mio Okeda, Ai Nakano, Taemi Hirshman, Michael F. Goodyear, Laurie J. Fujii, Nobuharu L. |
| spellingShingle |
Manabe, Yasuko Miyatake, Shouta Takagi, Mayumi Nakamura, Mio Okeda, Ai Nakano, Taemi Hirshman, Michael F. Goodyear, Laurie J. Fujii, Nobuharu L. Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes |
| author_facet |
Manabe, Yasuko Miyatake, Shouta Takagi, Mayumi Nakamura, Mio Okeda, Ai Nakano, Taemi Hirshman, Michael F. Goodyear, Laurie J. Fujii, Nobuharu L. |
| author_sort |
Manabe, Yasuko |
| title |
Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes |
| title_short |
Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes |
| title_full |
Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes |
| title_fullStr |
Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes |
| title_full_unstemmed |
Characterization of an Acute Muscle Contraction Model Using Cultured C2C12 Myotubes |
| title_sort |
characterization of an acute muscle contraction model using cultured c2c12 myotubes |
| description |
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. |
| publisher |
Public Library of Science |
| publishDate |
2012 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3534077/ |
| _version_ |
1611943913479733248 |