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Simulated microgravity attenuates myogenic differentiation via epigenetic regulations

The molecular mechanisms involved in myogenic differentiation are relatively well-known. Myogenic differentiation is regulated by the sequential activation of the basic helix-loop-helix myogenic regulatory transcription factors (MRFs), and biomechanical signals play an important role in the regulati...

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Autores principales: Furukawa, Takuma, Tanimoto, Keiji, Fukazawa, Takahiro, Imura, Takeshi, Kawahara, Yumi, Yuge, Louis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5966377/
https://www.ncbi.nlm.nih.gov/pubmed/29845109
http://dx.doi.org/10.1038/s41526-018-0045-0
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author Furukawa, Takuma
Tanimoto, Keiji
Fukazawa, Takahiro
Imura, Takeshi
Kawahara, Yumi
Yuge, Louis
author_facet Furukawa, Takuma
Tanimoto, Keiji
Fukazawa, Takahiro
Imura, Takeshi
Kawahara, Yumi
Yuge, Louis
author_sort Furukawa, Takuma
collection PubMed
description The molecular mechanisms involved in myogenic differentiation are relatively well-known. Myogenic differentiation is regulated by the sequential activation of the basic helix-loop-helix myogenic regulatory transcription factors (MRFs), and biomechanical signals play an important role in the regulation of myogenesis. In this study, we sought to determine whether simulated microgravity culture using Gravite(®) may affect myoblast differentiation and expression of MRF genes. Although rat myoblasts, L6 cells were differentiated to myotubes in an incubation period-dependent manner, myogenesis of L6 cells was significantly attenuated under simulated microgravity (10(-3)G) conditions. Real-time Reverse transcription polymerase chain reaction (RT-PCR) showed that expressions of Myog, Myf6, Mef2c, Des, and Ckm under 1 G conditions increase in an incubation period-dependent manner, and that Myod1 expression was specifically observed to increase transiently in the early phase. However, expressions of Myod1 and Myog were significantly inhibited under simulated microgravity conditions. To clarify the molecular mechanisms, L6 cells were treated with 5-AzaC, and further incubated with differentiation medium under 1 G or 10(−3) G conditions. The results showed differences in expression levels of Myod1, Myog, and, as well as those of myotube thickness between 1 G and 10(−3) G conditions, completely disappeared in this experimental condition. Modified HpaII tiny fragment enrichment by ligation-mediated PCR (HELP)-assay showed that kinetic changes of DNA methylation status were attenuated in simulated microgravity conditions. These results indicate that microgravity regulates myogenesis and Myod1 expression by controlling DNA methylation.
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spelling pubmed-59663772018-05-29 Simulated microgravity attenuates myogenic differentiation via epigenetic regulations Furukawa, Takuma Tanimoto, Keiji Fukazawa, Takahiro Imura, Takeshi Kawahara, Yumi Yuge, Louis NPJ Microgravity Article The molecular mechanisms involved in myogenic differentiation are relatively well-known. Myogenic differentiation is regulated by the sequential activation of the basic helix-loop-helix myogenic regulatory transcription factors (MRFs), and biomechanical signals play an important role in the regulation of myogenesis. In this study, we sought to determine whether simulated microgravity culture using Gravite(®) may affect myoblast differentiation and expression of MRF genes. Although rat myoblasts, L6 cells were differentiated to myotubes in an incubation period-dependent manner, myogenesis of L6 cells was significantly attenuated under simulated microgravity (10(-3)G) conditions. Real-time Reverse transcription polymerase chain reaction (RT-PCR) showed that expressions of Myog, Myf6, Mef2c, Des, and Ckm under 1 G conditions increase in an incubation period-dependent manner, and that Myod1 expression was specifically observed to increase transiently in the early phase. However, expressions of Myod1 and Myog were significantly inhibited under simulated microgravity conditions. To clarify the molecular mechanisms, L6 cells were treated with 5-AzaC, and further incubated with differentiation medium under 1 G or 10(−3) G conditions. The results showed differences in expression levels of Myod1, Myog, and, as well as those of myotube thickness between 1 G and 10(−3) G conditions, completely disappeared in this experimental condition. Modified HpaII tiny fragment enrichment by ligation-mediated PCR (HELP)-assay showed that kinetic changes of DNA methylation status were attenuated in simulated microgravity conditions. These results indicate that microgravity regulates myogenesis and Myod1 expression by controlling DNA methylation. Nature Publishing Group UK 2018-05-23 /pmc/articles/PMC5966377/ /pubmed/29845109 http://dx.doi.org/10.1038/s41526-018-0045-0 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Furukawa, Takuma
Tanimoto, Keiji
Fukazawa, Takahiro
Imura, Takeshi
Kawahara, Yumi
Yuge, Louis
Simulated microgravity attenuates myogenic differentiation via epigenetic regulations
title Simulated microgravity attenuates myogenic differentiation via epigenetic regulations
title_full Simulated microgravity attenuates myogenic differentiation via epigenetic regulations
title_fullStr Simulated microgravity attenuates myogenic differentiation via epigenetic regulations
title_full_unstemmed Simulated microgravity attenuates myogenic differentiation via epigenetic regulations
title_short Simulated microgravity attenuates myogenic differentiation via epigenetic regulations
title_sort simulated microgravity attenuates myogenic differentiation via epigenetic regulations
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5966377/
https://www.ncbi.nlm.nih.gov/pubmed/29845109
http://dx.doi.org/10.1038/s41526-018-0045-0
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