Profiling of adenine‐derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide‐induced cell stress
Abstract Cytokinins (CTKs) are a diverse collection of evolutionarily conserved adenine‐derived signaling molecules classically studied as phytohormones; however, their roles and production have been less studied in mammalian systems. Skeletal muscles are sensitive to cellular cues such as inflammat...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-12-01
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| Series: | Physiological Reports |
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| Online Access: | https://doi.org/10.14814/phy2.15870 |
| _version_ | 1797389353363701760 |
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| author | Stephanie W. Tobin Dev Seneviratne Lorna Phan Mark Seegobin Alexander L. Rico Beth Westby Anna Kisiala Sanela Martic Craig R. Brunetti R. J. Neil Emery |
| author_facet | Stephanie W. Tobin Dev Seneviratne Lorna Phan Mark Seegobin Alexander L. Rico Beth Westby Anna Kisiala Sanela Martic Craig R. Brunetti R. J. Neil Emery |
| author_sort | Stephanie W. Tobin |
| collection | DOAJ |
| description | Abstract Cytokinins (CTKs) are a diverse collection of evolutionarily conserved adenine‐derived signaling molecules classically studied as phytohormones; however, their roles and production have been less studied in mammalian systems. Skeletal muscles are sensitive to cellular cues such as inflammation and in response, alter their secretome to regulate the muscle stem cell and myofiber niche. Using cultured C2C12 muscle cells, we profiled CTK levels to understand (1) whether CTKs are part of the muscle secretome and (2) whether CTKs are responsive to cellular stress. To induce cellular stress, C2C12 myotubes were treated with lipopolysaccharides (LPS) for 24 h and then media and cell fractions were collected for ultra high‐performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC‐(ESI+)‐HRMS/MS) for metabolomics and CTK profiling. Across LPS‐treated and control cells, 11 CTKs were detected in the extracellular space while 6 were detected intracellularly. We found that muscle cells are enriched in isopentenyladenine (iP) species (from free base, riboside to nucleotide forms), and that extracellular levels are increased after LPS treatment. Our study establishes that muscle cells express various forms of CTKs, and that CTK levels are responsive to LPS‐induced cell stress, suggesting a role for CTKs in intra‐ and extracellular signaling of mammalian cells. |
| first_indexed | 2024-03-08T22:54:38Z |
| format | Article |
| id | doaj.art-91969bbed732491695458e7130372605 |
| institution | Directory Open Access Journal |
| issn | 2051-817X |
| language | English |
| last_indexed | 2024-03-08T22:54:38Z |
| publishDate | 2023-12-01 |
| publisher | Wiley |
| record_format | Article |
| series | Physiological Reports |
| spelling | doaj.art-91969bbed732491695458e71303726052023-12-16T06:30:46ZengWileyPhysiological Reports2051-817X2023-12-011123n/an/a10.14814/phy2.15870Profiling of adenine‐derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide‐induced cell stressStephanie W. Tobin0Dev Seneviratne1Lorna Phan2Mark Seegobin3Alexander L. Rico4Beth Westby5Anna Kisiala6Sanela Martic7Craig R. Brunetti8R. J. Neil Emery9Department of Biology Trent University Peterborough Ontario CanadaEnvironmental and Life Sciences Graduate Program Trent University Peterborough Ontario CanadaDepartment of Biology Trent University Peterborough Ontario CanadaDepartment of Biology Trent University Peterborough Ontario CanadaDepartment of Biology Trent University Peterborough Ontario CanadaDepartment of Biology Trent University Peterborough Ontario CanadaDepartment of Biology Trent University Peterborough Ontario CanadaEnvironmental and Life Sciences Graduate Program Trent University Peterborough Ontario CanadaDepartment of Biology Trent University Peterborough Ontario CanadaDepartment of Biology Trent University Peterborough Ontario CanadaAbstract Cytokinins (CTKs) are a diverse collection of evolutionarily conserved adenine‐derived signaling molecules classically studied as phytohormones; however, their roles and production have been less studied in mammalian systems. Skeletal muscles are sensitive to cellular cues such as inflammation and in response, alter their secretome to regulate the muscle stem cell and myofiber niche. Using cultured C2C12 muscle cells, we profiled CTK levels to understand (1) whether CTKs are part of the muscle secretome and (2) whether CTKs are responsive to cellular stress. To induce cellular stress, C2C12 myotubes were treated with lipopolysaccharides (LPS) for 24 h and then media and cell fractions were collected for ultra high‐performance liquid chromatography tandem mass spectrometry with electrospray ionization (UHPLC‐(ESI+)‐HRMS/MS) for metabolomics and CTK profiling. Across LPS‐treated and control cells, 11 CTKs were detected in the extracellular space while 6 were detected intracellularly. We found that muscle cells are enriched in isopentenyladenine (iP) species (from free base, riboside to nucleotide forms), and that extracellular levels are increased after LPS treatment. Our study establishes that muscle cells express various forms of CTKs, and that CTK levels are responsive to LPS‐induced cell stress, suggesting a role for CTKs in intra‐ and extracellular signaling of mammalian cells.https://doi.org/10.14814/phy2.15870cytokininmusclemyokine |
| spellingShingle | Stephanie W. Tobin Dev Seneviratne Lorna Phan Mark Seegobin Alexander L. Rico Beth Westby Anna Kisiala Sanela Martic Craig R. Brunetti R. J. Neil Emery Profiling of adenine‐derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide‐induced cell stress Physiological Reports cytokinin muscle myokine |
| title | Profiling of adenine‐derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide‐induced cell stress |
| title_full | Profiling of adenine‐derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide‐induced cell stress |
| title_fullStr | Profiling of adenine‐derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide‐induced cell stress |
| title_full_unstemmed | Profiling of adenine‐derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide‐induced cell stress |
| title_short | Profiling of adenine‐derived signaling molecules, cytokinins, in myotubes reveals fluctuations in response to lipopolysaccharide‐induced cell stress |
| title_sort | profiling of adenine derived signaling molecules cytokinins in myotubes reveals fluctuations in response to lipopolysaccharide induced cell stress |
| topic | cytokinin muscle myokine |
| url | https://doi.org/10.14814/phy2.15870 |
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