Divergent regulation of myotube formation and gene expression by E2 and EPA during in-vitro differentiation of C2C12 myoblasts
Journal article
Lacham-Kaplan, Orly, Camera, Donny and Hawley, John. (2020). Divergent regulation of myotube formation and gene expression by E2 and EPA during in-vitro differentiation of C2C12 myoblasts. International Journal of Molecular Sciences. 21(3), pp. 1-18. https://doi.org/10.3390/ijms21030745
Authors | Lacham-Kaplan, Orly, Camera, Donny and Hawley, John |
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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 50μM eicosapentaenoic acid (EPA) independently or combined, for 0–24 h or 0–120 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 α (Erα) 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 < 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. |
Keywords | 17β-estradiol; n-3PUFA; eicosapentaenoic acid; transcriptome; C2C12; myogenesis |
Year | 2020 |
Journal | International Journal of Molecular Sciences |
Journal citation | 21 (3), pp. 1-18 |
Publisher | Multidisciplinary Digital Publishing Institute (MDPI AG) |
ISSN | 1661-6596 |
Digital Object Identifier (DOI) | https://doi.org/10.3390/ijms21030745 |
Scopus EID | 2-s2.0-85078277542 |
Open access | Published as ‘gold’ (paid) open access |
Research or scholarly | Research |
Page range | 1-18 |
Publisher's version | License File Access Level Open |
Output status | Published |
Publication dates | |
Online | 23 Jan 2020 |
Publication process dates | |
Accepted | 21 Jan 2020 |
Deposited | 13 Apr 2021 |
https://acuresearchbank.acu.edu.au/item/8vwy2/divergent-regulation-of-myotube-formation-and-gene-expression-by-e2-and-epa-during-in-vitro-differentiation-of-c2c12-myoblasts
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Publisher's version
OA_Lacham-Kaplan_2020_Divergent_regulation_of_myotube_formation_and.pdf | |
License: CC BY 4.0 | |
File access level: Open |
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