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Divergent Regulation of Myotube Formation and Gene Expression by E2 and EPA during In-Vitro Differentiation of C2C12 Myoblasts
- Source :
- International Journal of Molecular Sciences, Vol 21, Iss 3, p 745 (2020), International Journal of Molecular Sciences, Volume 21, Issue 3
- Publication Year :
- 2020
- Publisher :
- MDPI AG, 2020.
-
Abstract
- Estrogen (E2) and polyunsaturated fatty acids (n-3PUFA) supplements independently support general wellbeing and enhance muscle regeneration in-vivo and myotube formation in-vitro. However, the combined effect of E2 and n-3PUFA on myoblast differentiation is not known. The purpose of the study was to identify whether E2 and n-3PUFA possess a synergistic effect on in-vitro myogenesis. Mouse C2C12 myoblasts, a reliable model to reiterate myogenic events in-vitro, were treated with 10nM E2 and 50&mu<br />M eicosapentaenoic acid (EPA) independently or combined, for 0&ndash<br />24 h or 0&ndash<br />120 h during differentiation. Immunofluorescence, targeted qPCR and next generation sequencing (NGS) were used to characterize morphological changes and differential expression of key genes involved in the regulation of myogenesis and muscle function pathways. E2 increased estrogen receptor &alpha<br />(Er&alpha<br />) and the expression of the mitogen-activated protein kinase 11 (Mapk11) within 1 h of treatment and improved myoblast differentiation and myotube formation. A significant reduction (p &lt<br />0.001) in myotube formation and in the expression of myogenic regulatory factors Mrfs (MyoD, Myog and Myh1) and the myoblast fusion related gene, Tmem8c, was observed in the presence of EPA and the combined E2/EPA treatment. Additionally, EPA treatment at 48 h of differentiation inhibited the majority of genes associated with the myogenic and striated muscle contraction pathways. In conclusion, EPA and E2 had no synergistic effect on myotube formation in-vitro. Independently, EPA inhibited myoblast differentiation and overrides the stimulatory effect of E2 when used in combination with E2.
- Subjects :
- eicosapentaenoic acid
Muscle Fibers, Skeletal
Muscle Proteins
Muscle Development
MyoD
DNA Glycosylases
Myoblasts
lcsh:Chemistry
Mice
Myoblast fusion
Mitogen-Activated Protein Kinase 11
Myocyte
lcsh:QH301-705.5
Spectroscopy
Myogenesis
Chemistry
n-3pufa
High-Throughput Nucleotide Sequencing
Drug Synergism
General Medicine
Striated muscle contraction
musculoskeletal system
Computer Science Applications
Cell biology
c2c12
Fatty Acids, Unsaturated
Myogenin
myogenesis
C2C12
Signal Transduction
Article
Catalysis
Cell Line
Inorganic Chemistry
Animals
MAPK11
Physical and Theoretical Chemistry
Molecular Biology
MyoD Protein
Organic Chemistry
Estrogen Receptor alpha
17β-estradiol
Membrane Proteins
Estrogens
Gene Ontology
Gene Expression Regulation
lcsh:Biology (General)
lcsh:QD1-999
Myogenic regulatory factors
transcriptome
Subjects
Details
- ISSN :
- 14220067
- Volume :
- 21
- Database :
- OpenAIRE
- Journal :
- International Journal of Molecular Sciences
- Accession number :
- edsair.doi.dedup.....749cf395dd1c0ee955e82c165198ac84