|
| Status |
Public on Jan 01, 2012 |
| Title |
Effect of 5-azacytidine on gene expression in C2C12 myoblasts |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
|
| Summary |
Mesenchymal progenitor cells can be differentiated in vitro into myotubes that exhibit many characteristic features of primary mammalian skeletal muscle fibers. However, in general, they do not show the functional excitation-contraction coupling or the striated sarcomere arrangement typical of mature myofibers. Epigenetic modifications have been shown to play a key role in regulating the progressional changes in transcription necessary for muscle differentiation. In this study, we demonstrate that treatment of murine C2C12 mesenchymal progenitor cells with 10 µM of the DNA methylation inhibitor 5-azacytidine (5AC) promotes myogenesis, resulting in myotubes with enhanced maturity as compared to untreated myotubes. Specifically, 5AC treatment resulted in the upregulation of muscle genes at the myoblast stage while at later stages nearly 50 % of the 5AC-treated myotubes displayed a mature, well-defined sarcomere organization as well as spontaneous contractions that coincided with action potentials and intracellular calcium transients. Both the percentage of striated myotubes and their contractile activity could be inhibited by 20 nM TTX, 10 µM ryanodine and 100 µM nifedipine, suggesting that action potential-induced calcium transients are responsible for these characteristics. Our data suggest that genomic demethylation induced by 5AC overcomes an epigenetic barrier that prevents untreated C2C12 myotubes from reaching full maturity.
|
| |
|
| Overall design |
C2C12 cells were plated at 1500 cells/cm2 (day -1), cultured for 1 day in DMEM 10%NCS, then treated (d0) with or without 10uM 5-azacytidine (5AC) for an additional 3 days. RNA was extracted on day 3 and hybridized to GeneChip Mouse Genome 430 2.0 array (Affymetrix).
|
| |
|
| Contributor(s) |
Hupkes M, Sotoca AM, van Someren EP, van Ravestein-van Os RI, Bauerschmidt S, van Zoelen EJ, Dechering KJ |
| Citation(s) |
21795504 |
| |
| Submission date |
Jun 24, 2011 |
| Last update date |
Feb 11, 2019 |
| Contact name |
Marlinda Hupkes |
| E-mail(s) |
[email protected]
|
| Phone |
+31(0)243652519
|
| Organization name |
Radboud University Nijmegen
|
| Department |
Cell and Applied Biology
|
| Street address |
Heyendaalseweg 135
|
| City |
Nijmegen |
| ZIP/Postal code |
6525 AJ |
| Country |
Netherlands |
| |
|
| Platforms (1) |
| GPL1261 |
[Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array |
|
| Samples (6)
|
|
| Relations |
| BioProject |
PRJNA143943 |
Less...
More...