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Elevation of phosphate levels impairs skeletal myoblast differentiation
Hyperphosphatemic conditions such as chronic kidney disease are associated with severe muscle wasting and impaired life quality. While regeneration of muscle tissue is known to be reliant on recruitment of myogenic progenitor cells, the effects of elevated phosphate loads on this process have not be...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7584532/ https://www.ncbi.nlm.nih.gov/pubmed/32725423 http://dx.doi.org/10.1007/s00441-020-03254-1 |
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author | Raimann, Adalbert Dangl, Alexander Javanmardi, Alireza Greber-Platzer, Susanne Egerbacher, Monika Pietschmann, Peter Haeusler, Gabriele |
author_facet | Raimann, Adalbert Dangl, Alexander Javanmardi, Alireza Greber-Platzer, Susanne Egerbacher, Monika Pietschmann, Peter Haeusler, Gabriele |
author_sort | Raimann, Adalbert |
collection | PubMed |
description | Hyperphosphatemic conditions such as chronic kidney disease are associated with severe muscle wasting and impaired life quality. While regeneration of muscle tissue is known to be reliant on recruitment of myogenic progenitor cells, the effects of elevated phosphate loads on this process have not been investigated in detail so far. This study aims to clarify the direct effects of hyperphosphatemic conditions on skeletal myoblast differentiation in a murine in vitro model. C2C12 murine muscle progenitor cells were supplemented with phosphate concentrations resembling moderate to severe hyperphosphatemia (1.4–2.9 mmol/l). Phosphate-induced effects were quantified by RT-PCR and immunoblotting. Immunohistochemistry was performed to count nuclear positive cells under treatment. Cell viability and metabolic activity were assessed by XTT and BrdU incorporation assays. Inorganic phosphate directly induced ERK-phosphorylation in pre-differentiated C2C12 myoblast cells. While phosphate concentrations resembling the upper normal range significantly reduced Myogenin expression (− 22.5%, p = 0.015), severe hyperphosphatemic conditions further impaired differentiation (Myogenin − 61.0%, p < 0.0001; MyoD − 51.0%; p < 0.0001). Analogue effects were found on the protein level (Myogenin − 42.0%, p = 0.004; MyoD − 25.7%, p = 0.002). ERK inhibition strongly attenuated phosphate-induced effects on Myogenin expression (p = 0.002). Metabolic activity was unaffected by the treatments. Our data point to a phosphate-induced inhibition of myoblast differentiation without effects on cell viability. Serum phosphate levels as low as the upper normal serum range significantly impaired marker gene expression in vitro. Investigation of cellular effects of hyperphosphatemia may help to better define serum cutoffs and modify existing treatment approaches of phosphate binders, especially in patients at risk of sarcopenia. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00441-020-03254-1) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-7584532 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-75845322020-10-27 Elevation of phosphate levels impairs skeletal myoblast differentiation Raimann, Adalbert Dangl, Alexander Javanmardi, Alireza Greber-Platzer, Susanne Egerbacher, Monika Pietschmann, Peter Haeusler, Gabriele Cell Tissue Res Short Communication Hyperphosphatemic conditions such as chronic kidney disease are associated with severe muscle wasting and impaired life quality. While regeneration of muscle tissue is known to be reliant on recruitment of myogenic progenitor cells, the effects of elevated phosphate loads on this process have not been investigated in detail so far. This study aims to clarify the direct effects of hyperphosphatemic conditions on skeletal myoblast differentiation in a murine in vitro model. C2C12 murine muscle progenitor cells were supplemented with phosphate concentrations resembling moderate to severe hyperphosphatemia (1.4–2.9 mmol/l). Phosphate-induced effects were quantified by RT-PCR and immunoblotting. Immunohistochemistry was performed to count nuclear positive cells under treatment. Cell viability and metabolic activity were assessed by XTT and BrdU incorporation assays. Inorganic phosphate directly induced ERK-phosphorylation in pre-differentiated C2C12 myoblast cells. While phosphate concentrations resembling the upper normal range significantly reduced Myogenin expression (− 22.5%, p = 0.015), severe hyperphosphatemic conditions further impaired differentiation (Myogenin − 61.0%, p < 0.0001; MyoD − 51.0%; p < 0.0001). Analogue effects were found on the protein level (Myogenin − 42.0%, p = 0.004; MyoD − 25.7%, p = 0.002). ERK inhibition strongly attenuated phosphate-induced effects on Myogenin expression (p = 0.002). Metabolic activity was unaffected by the treatments. Our data point to a phosphate-induced inhibition of myoblast differentiation without effects on cell viability. Serum phosphate levels as low as the upper normal serum range significantly impaired marker gene expression in vitro. Investigation of cellular effects of hyperphosphatemia may help to better define serum cutoffs and modify existing treatment approaches of phosphate binders, especially in patients at risk of sarcopenia. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00441-020-03254-1) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2020-07-28 2020 /pmc/articles/PMC7584532/ /pubmed/32725423 http://dx.doi.org/10.1007/s00441-020-03254-1 Text en © The Author(s) 2020 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Short Communication Raimann, Adalbert Dangl, Alexander Javanmardi, Alireza Greber-Platzer, Susanne Egerbacher, Monika Pietschmann, Peter Haeusler, Gabriele Elevation of phosphate levels impairs skeletal myoblast differentiation |
title | Elevation of phosphate levels impairs skeletal myoblast differentiation |
title_full | Elevation of phosphate levels impairs skeletal myoblast differentiation |
title_fullStr | Elevation of phosphate levels impairs skeletal myoblast differentiation |
title_full_unstemmed | Elevation of phosphate levels impairs skeletal myoblast differentiation |
title_short | Elevation of phosphate levels impairs skeletal myoblast differentiation |
title_sort | elevation of phosphate levels impairs skeletal myoblast differentiation |
topic | Short Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7584532/ https://www.ncbi.nlm.nih.gov/pubmed/32725423 http://dx.doi.org/10.1007/s00441-020-03254-1 |
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