Epigenetic Modifier Supplementation Improves Mitochondrial Respiration and Growth Rates and Alters DNA Methylation of Bovine Embryonic Fibroblast Cells Cultured in Divergent Energy Supply
Epigenetic modifiers (EM; methionine, choline, folate, and vitamin B12) are important for early embryonic development due to their roles as methyl donors or cofactors in methylation reactions. Additionally, they are essential for the synthesis of nucleotides, polyamines, redox equivalents, and energ...
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Frontiers Media S.A.
2022-02-01
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| Series: | Frontiers in Genetics |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fgene.2022.812764/full |
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| author | Matthew S. Crouse Joel S. Caton Kate J. Claycombe-Larson Wellison J. S. Diniz Amanda K. Lindholm-Perry Lawrence P. Reynolds Carl R. Dahlen Pawel P. Borowicz Alison K. Ward |
| author_facet | Matthew S. Crouse Joel S. Caton Kate J. Claycombe-Larson Wellison J. S. Diniz Amanda K. Lindholm-Perry Lawrence P. Reynolds Carl R. Dahlen Pawel P. Borowicz Alison K. Ward |
| author_sort | Matthew S. Crouse |
| collection | DOAJ |
| description | Epigenetic modifiers (EM; methionine, choline, folate, and vitamin B12) are important for early embryonic development due to their roles as methyl donors or cofactors in methylation reactions. Additionally, they are essential for the synthesis of nucleotides, polyamines, redox equivalents, and energy metabolites. Despite their importance, investigation into the supplementation of EM in ruminants has been limited to one or two epigenetic modifiers. Like all biochemical pathways, one-carbon metabolism needs to be stoichiometrically balanced. Thus, we investigated the effects of supplementing four EM encompassing the methionine–folate cycle on bovine embryonic fibroblast growth, mitochondrial function, and DNA methylation. We hypothesized that EM supplemented to embryonic fibroblasts cultured in divergent glucose media would increase mitochondrial respiration and cell growth rate and alter DNA methylation as reflected by changes in the gene expression of enzymes involved in methylation reactions, thereby improving the growth parameters beyond Control treated cells. Bovine embryonic fibroblast cells were cultured in Eagle’s minimum essential medium with 1 g/L glucose (Low) or 4.5 g/L glucose (High). The control medium contained no additional OCM, whereas the treated media contained supplemented EM at 2.5, 5, and 10 times (×2.5, ×5, and ×10, respectively) the control media, except for methionine (limited to ×2). Therefore, the experimental design was a 2 (levels of glucose) × 4 (levels of EM) factorial arrangement of treatments. Cells were passaged three times in their respective treatment media before analysis for growth rate, cell proliferation, mitochondrial respiration, transcript abundance of methionine–folate cycle enzymes, and DNA methylation by reduced-representation bisulfite sequencing. Total cell growth was greatest in High ×10 and mitochondrial maximal respiration, and reserve capacity was greatest (p < 0.01) for High ×2.5 and ×10 compared with all other treatments. In Low cells, the total growth rate, mitochondrial maximal respiration, and reserve capacity increased quadratically to 2.5 and ×5 and decreased to control levels at ×10. The biological processes identified due to differential methylation included the positive regulation of GTPase activity, molecular function, protein modification processes, phosphorylation, and metabolic processes. These data are interpreted to imply that EM increased the growth rate and mitochondrial function beyond Control treated cells in both Low and High cells, which may be due to changes in the methylation of genes involved with growth and energy metabolism. |
| first_indexed | 2024-12-19T12:33:14Z |
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| id | doaj.art-74e7ef77a6f341dcb7c009aca2695405 |
| institution | Directory Open Access Journal |
| issn | 1664-8021 |
| language | English |
| last_indexed | 2024-12-19T12:33:14Z |
| publishDate | 2022-02-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Genetics |
| spelling | doaj.art-74e7ef77a6f341dcb7c009aca26954052022-12-21T20:21:13ZengFrontiers Media S.A.Frontiers in Genetics1664-80212022-02-011310.3389/fgene.2022.812764812764Epigenetic Modifier Supplementation Improves Mitochondrial Respiration and Growth Rates and Alters DNA Methylation of Bovine Embryonic Fibroblast Cells Cultured in Divergent Energy SupplyMatthew S. Crouse0Joel S. Caton1Kate J. Claycombe-Larson2Wellison J. S. Diniz3Amanda K. Lindholm-Perry4Lawrence P. Reynolds5Carl R. Dahlen6Pawel P. Borowicz7Alison K. Ward8USDA, ARS, U.S. Meat Animal Research Center, Clay Center, NE, United StatesDepartment of Animal Sciences, North Dakota State University, Fargo, ND, United StatesUSDA, ARS, Grand Forks Human Nutrition Research Center, Grand Forks, ND, United StatesDepartment of Animal Sciences, Auburn University, Auburn, AL, United StatesUSDA, ARS, U.S. Meat Animal Research Center, Clay Center, NE, United StatesDepartment of Animal Sciences, North Dakota State University, Fargo, ND, United StatesDepartment of Animal Sciences, North Dakota State University, Fargo, ND, United StatesDepartment of Animal Sciences, North Dakota State University, Fargo, ND, United StatesDepartment of Animal Sciences, North Dakota State University, Fargo, ND, United StatesEpigenetic modifiers (EM; methionine, choline, folate, and vitamin B12) are important for early embryonic development due to their roles as methyl donors or cofactors in methylation reactions. Additionally, they are essential for the synthesis of nucleotides, polyamines, redox equivalents, and energy metabolites. Despite their importance, investigation into the supplementation of EM in ruminants has been limited to one or two epigenetic modifiers. Like all biochemical pathways, one-carbon metabolism needs to be stoichiometrically balanced. Thus, we investigated the effects of supplementing four EM encompassing the methionine–folate cycle on bovine embryonic fibroblast growth, mitochondrial function, and DNA methylation. We hypothesized that EM supplemented to embryonic fibroblasts cultured in divergent glucose media would increase mitochondrial respiration and cell growth rate and alter DNA methylation as reflected by changes in the gene expression of enzymes involved in methylation reactions, thereby improving the growth parameters beyond Control treated cells. Bovine embryonic fibroblast cells were cultured in Eagle’s minimum essential medium with 1 g/L glucose (Low) or 4.5 g/L glucose (High). The control medium contained no additional OCM, whereas the treated media contained supplemented EM at 2.5, 5, and 10 times (×2.5, ×5, and ×10, respectively) the control media, except for methionine (limited to ×2). Therefore, the experimental design was a 2 (levels of glucose) × 4 (levels of EM) factorial arrangement of treatments. Cells were passaged three times in their respective treatment media before analysis for growth rate, cell proliferation, mitochondrial respiration, transcript abundance of methionine–folate cycle enzymes, and DNA methylation by reduced-representation bisulfite sequencing. Total cell growth was greatest in High ×10 and mitochondrial maximal respiration, and reserve capacity was greatest (p < 0.01) for High ×2.5 and ×10 compared with all other treatments. In Low cells, the total growth rate, mitochondrial maximal respiration, and reserve capacity increased quadratically to 2.5 and ×5 and decreased to control levels at ×10. The biological processes identified due to differential methylation included the positive regulation of GTPase activity, molecular function, protein modification processes, phosphorylation, and metabolic processes. These data are interpreted to imply that EM increased the growth rate and mitochondrial function beyond Control treated cells in both Low and High cells, which may be due to changes in the methylation of genes involved with growth and energy metabolism.https://www.frontiersin.org/articles/10.3389/fgene.2022.812764/fullcell growthDNA methylationembryonic fibroblastsmitochondrial respirationone-carbon metabolism |
| spellingShingle | Matthew S. Crouse Joel S. Caton Kate J. Claycombe-Larson Wellison J. S. Diniz Amanda K. Lindholm-Perry Lawrence P. Reynolds Carl R. Dahlen Pawel P. Borowicz Alison K. Ward Epigenetic Modifier Supplementation Improves Mitochondrial Respiration and Growth Rates and Alters DNA Methylation of Bovine Embryonic Fibroblast Cells Cultured in Divergent Energy Supply Frontiers in Genetics cell growth DNA methylation embryonic fibroblasts mitochondrial respiration one-carbon metabolism |
| title | Epigenetic Modifier Supplementation Improves Mitochondrial Respiration and Growth Rates and Alters DNA Methylation of Bovine Embryonic Fibroblast Cells Cultured in Divergent Energy Supply |
| title_full | Epigenetic Modifier Supplementation Improves Mitochondrial Respiration and Growth Rates and Alters DNA Methylation of Bovine Embryonic Fibroblast Cells Cultured in Divergent Energy Supply |
| title_fullStr | Epigenetic Modifier Supplementation Improves Mitochondrial Respiration and Growth Rates and Alters DNA Methylation of Bovine Embryonic Fibroblast Cells Cultured in Divergent Energy Supply |
| title_full_unstemmed | Epigenetic Modifier Supplementation Improves Mitochondrial Respiration and Growth Rates and Alters DNA Methylation of Bovine Embryonic Fibroblast Cells Cultured in Divergent Energy Supply |
| title_short | Epigenetic Modifier Supplementation Improves Mitochondrial Respiration and Growth Rates and Alters DNA Methylation of Bovine Embryonic Fibroblast Cells Cultured in Divergent Energy Supply |
| title_sort | epigenetic modifier supplementation improves mitochondrial respiration and growth rates and alters dna methylation of bovine embryonic fibroblast cells cultured in divergent energy supply |
| topic | cell growth DNA methylation embryonic fibroblasts mitochondrial respiration one-carbon metabolism |
| url | https://www.frontiersin.org/articles/10.3389/fgene.2022.812764/full |
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