Remodeling of skeletal muscle myosin metabolic states in hibernating mammals
- Department of Biomedical Sciences, University of Copenhagen, Denmark
- Centre for Human and Applied Physiological Sciences, Faculty of Life Sciences & Medicine, King’s College London, United Kingdom
- Department of Cellular and Molecular Medicine, University of Copenhagen, Denmark
- Department of Computational Medicine and Bioinformatics, University of Michigan, United States
- Spring-8, Japan Synchrotron Radiation Research Institute, Japan
- Institute of Physiology II, University of Muenster, Germany
- Accelerated Muscle Biotechnologies Consultants, United States
- Biosfer Teslab, Spain
- Department of Clinical Medicine, Faculty of Health, Aarhus University, Denmark
- Faculty of Health, Department of Cardiology, Örebro University, Sweden
- Energetics Lab, Department of Biology, Northern Michigan University, United States
- Research Institute of Wildlife Ecology, Department of Interdisciplinary Life Sciences, University of Veterinary Medicine Vienna, Austria
- Department of Biology, University of Western Ontario, Canada
- Center for Transformative Research in Metabolism, Institute of Arctic Biology, University of Alaska Fairbanks, United States
- Department of Zoology, University of British Columbia, Canada
Figures
Figure 1
Myosin dynamics and myosin ATP consumption is unchanged in Ursus arctos and Ursus americanus during hibernation.
(A) Representative fluorescence mant-ATP decays from single muscle fibers isolated from Ursus arctos skeletal muscle measured over 300 s. (B–C) Percentage of myosin heads in the P1/DRX (B) or P2/SRX …
Figure 2
Myosin ATP turnover lifetime is reduced during hibernation in small hibernators, Eliomys quercinus and Ictidomys tridecemlineatus, resulting in an increase in myosin ATP consumption at ambient temperatures.
(A–B) Percentage of myosin heads in the P1/DRX (A) or P2/SRX (B) from E. quercinus single muscle fibers obtained during active, interbout arousal (IBA) or torpor periods. Values were separated based …
Figure 3
X-ray diffraction experiments of skeletal muscle from Ictidomys tridecemlineaus demonstrate changes in M6 myosin meridional spacing during torpor.
(A) Representative X-ray diffraction recordings from permeabilized skeletal muscle bundles from Ictidomys tridecemlineatus from summer active (SA), interbout arousal (IBA) and torpor. The M3 and M6 …
Figure 4 with 1 supplement
Myosin dynamics of Ictidomys tridecemlineatus are protected from temperature induced change during torpor, preventing an increase in myosin ATP consumption.
(A) T1 value in seconds denoting the ATP turnover lifetime of the DRX in I. tridecemlineatus at 8 °C. (B) T2 value in seconds denoting the ATP turnover lifetime in seconds of the SRX in I. …
Figure 4—figure supplement 1
Myosin ATP turnover lifetime is altered following exposure to cold temperature in MyHC-II muscle fibers from I. tridecemlineatus during active and IBA periods but not torpor.
(A–B) Percentage of myosin heads in the P1/DRX (A) or P2/SRX (B) from I. tridecemlineatus single muscle fibers obtained during summer active (SA), interbout arousal (IBA) or torpor periods at 8 °C. …
Figure 5 with 2 supplements
MYH2 protein in Ictidomys tridecemlineatus is hyper-phosphorylated during torpor, which is predicted to increase protein stability.
(A) Peptide mapping of differentiated phosphorylation sites upon MYH2 protein during SA, IBA and torpor periods. Heat map demonstrates all sites observed to be differentiated following the …
Figure 5—figure supplement 1
MYH7 protein phosphorylation and acetylation in U. arctos is relatively unchanged during winter periods.
(A) Peptide mapping of differentiated phosphorylation sites upon MYH7 protein during summer and winter periods. Heat map demonstrates all sites observed to be differentiated following the …
Figure 5—figure supplement 2
MYH2 protein phosphorylation and acetylation in U.
arctos is altered during winter periods. (A) Peptide mapping of differentiated phosphorylation sites upon MYH2 protein during summer and winter periods. Heat map demonstrates all sites observed to …
Figure 6 with 5 supplements
Global proteome analysis demonstrates changes to metabolic and sarcomeric changes in skeletal muscle fibers from Ictidomys tridecemlineatus during IBA and torpor.
(A) Principal component analysis for all animals analyzed during SA, IBA and torpor periods. (B) Volcano plot displaying proteins which are differentially expressed during torpor vs active periods. …
Figure 6—figure supplement 1
All ontological clusters altered in I. tridecemlineatus in torpor vs SA periods.
(A) All upregulated pathways identified in Metascape in torpor vs SA periods. Pathways are listed in order of significance by -log10(p). (B) All downregulated pathways identified in Metascape in …
Figure 6—figure supplement 2
All ontological clusters altered in I. tridecemlineatus in IBA vs SA periods.
A. All upregulated pathways identified in Metascape in IBA vs SA periods. Pathways are listed in order of significance by -log10(p). (B) All downregulated pathways identified in Metascape in IBA vs …
Figure 6—figure supplement 3
Metabolite and lipid quantification of skeletal muscle from I.tridecemlineatus reveals a decrease in lipid levels during torpor.
(A) Quantification of metabolites from the skeletal muscle tissue of I. tridecemlineatus during SA, IBA, and torpor periods. Presented in µmol of metabolite per gram of tissue. (B) Quantification of …
Figure 6—figure supplement 4
Global proteome analysis of U.arctos skeletal muscle fibers reveal metabolic changes but not sarcomeric changes.
(A) Principal component analysis for all animals analyzed during summer and winter periods. (B) Volcano plot displaying proteins which are differentially expressed during winter vs summer periods. …
Figure 6—figure supplement 5
All ontological clusters altered in U.arctos in summer vs winter periods.
(A) All upregulated pathways identified in Metascape in winter vs summer periods. Pathways are listed in order of significance by -log10(p). (B) All downregulated pathways identified in Metascape in …
Additional files
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Supplementary file 1
Fiber type compositions from animals used for this study.
Table demonstrating the percentage of fibers which were analyzed during Mant-ATP chase assays that were either MyHC type I or MyHC type II. Data is presented as mean for each animal ± SD. One-way ANOVA was used to calculate significance between hibernation periods in I. tridecemlineatus and E. quercinus. Student’s t-test was used to calculate significant between hibernating periods in U. arctos and U. americanus and between MyHC type I and MyHC type II in all animals. ##=p < 0.01 vs MyHC type I in corresponding group. ####=p < 0.0001 vs MyHC type I in corresponding group. n=5 individual animals per group.
- https://cdn.elifesciences.org/articles/94616/elife-94616-supp1-v1.docx
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Supplementary file 2
Detailed protein lists for top five differentially expressed ontological clusters in I. tridecemlineatus in torpor vs SA periods.
A. Table details the list of proteins which were found to be differentially upregulated in torpor vs SA in each corresponding ontological cluster as identified by Metascape. Clusters are arranged by order of statistical significance. Proteins are listed in alphabetical order within each cluster. B. Table details the list of proteins which were found to be differentially downregulated in torpor vs SA in each corresponding ontological cluster as identified by Metascape. Clusters are arranged by order of statistical significance. Proteins are listed in alphabetical order within each cluster. n=5 individual animals per group.
- https://cdn.elifesciences.org/articles/94616/elife-94616-supp2-v1.docx
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Supplementary file 3
Detailed protein lists for top five differentially expressed ontological clusters in I. tridecemlineatus in IBA vs SA periods.
A. Table details the list of proteins which were found to be differentially upregulated in IBA vs SA in each corresponding ontological cluster as identified by Metascape. Clusters are arranged by order of statistical significance. Proteins are listed in alphabetical order within each cluster. B. Table details the list of proteins which were found to be differentially downregulated in IBA vs SA in each corresponding ontological cluster as identified by Metascape. Clusters are arranged by order of statistical significance. Proteins are listed in alphabetical order within each cluster. n=5 individual animals per group.
- https://cdn.elifesciences.org/articles/94616/elife-94616-supp3-v1.docx
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Supplementary file 4
Detailed protein lists for top five differentially expressed ontological clusters in U. arctos in winter vs summer periods.
A. Table details the list of proteins which were found to be differentially upregulated in winter vs summer in each corresponding ontological cluster as identified by Metascape. Clusters are arranged by order of statistical significance. Proteins are listed in alphabetical order within each cluster. B. Table details the list of proteins which were found to be differentially downregulated in winter vs summer in each corresponding ontological cluster as identified by Metascape. Clusters are arranged by order of statistical significance. Proteins are listed in alphabetical order within each cluster. n=5 individual animals per group.
- https://cdn.elifesciences.org/articles/94616/elife-94616-supp4-v1.docx
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MDAR checklist
- https://cdn.elifesciences.org/articles/94616/elife-94616-mdarchecklist1-v1.docx
