title: Skeletal muscle proteomics: carbohydrate metabolism oscillates with seasonal and torpor-arousal physiology of hibernation.
abstract: The physiology of small mammalian hibernators shifts profoundly over a year, from summer homeothermy to winter heterothermy. Torpor-arousal cycles define high-amplitude tissue activity fluctuations in winter, particularly for skeletal muscle, which contributes to the energetically demanding rewarming process via shivering. To better understand the biochemistry underlying summer-winter and torpor-arousal transitions, we applied two-dimensional gel electrophoresis coupled with liquid chromatography/mass spectrometry/mas spectrometry to the soluble proteins from hindlimb muscle of 13-lined ground squirrels (Ictidomys tridecemlineatus) in two summer and six winter states. Two hundred sixteen protein spots differed by sampled state. Significantly, intrawinter protein adjustment was a minor component of the dataset despite large discrepancies in muscle activity level among winter states; rather, the bulk of differences (127/138 unequivocally identified proteins spots) occurred between summer and winter. We did not detect any proteomic signatures of skeletal muscle atrophy in this hibernator nor any differential seasonal regulation of protein metabolism. Instead, adjustments to metabolic substrate preferences dominated the detected proteomic differences. Pathways of carbohydrate metabolism (glycolysis and gluconeogenesis) were summer enriched, whereas the winter proteome was enriched for fatty acid β-oxidation. Nevertheless, our data suggest that some reliance on carbohydrate reserves is maintained during winter. Phosphoglucomutase (PGM1), which reversibly prepares glucose subunits for either glycolysis or glycogenesis, showed apparent winter state-specific phosphorylation. PGM1 was phosphorylated during rewarming and dephosphorylated by interbout arousal, implying that glucose supplements lipid fuels during rewarming. This, along with winter elevation of TCA cycle enzymes, suggests that hindlimb muscles are primed for rapid energy production and that carbohydrates are an important fuel for shivering thermogenesis.
authors.affiliation: Dept. of Cell and Developmental Biology, PO Box 6511, MS 8010, Univ. of Colorado School of Medicine, Anschutz Medical Campus, Aurora, CO 80045, USA. allyson.hindle@ucdenver.edu
authors.forename: Allyson G
authors.initials: AG
authors.lastname: Hindle
authors.forename: Anis
authors.initials: A
authors.lastname: Karimpour-Fard
authors.forename: L Elaine
authors.initials: LE
authors.lastname: Epperson
authors.forename: Lawrence E
authors.initials: LE
authors.lastname: Hunter
authors.forename: Sandra L
authors.initials: SL
authors.lastname: Martin
mesh_terms: D000818:Animals; D001143:Arousal; D002853:Chromatography, Liquid; D002952:Citric Acid Cycle; D016000:Cluster Analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D005943:Gluconeogenesis; D006019:Glycolysis; D006605:Hibernation; D006614:Hindlimb; D050356:Lipid Metabolism; D013058:Mass Spectrometry; D009124:Muscle Proteins / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism*; D010766:Phosphorylation; D040901:Proteomics / Q000379:methods; D012589:Sciuridae; D012621:Seasons; D022722:Thermogenesis
chemical_list: D009124:Muscle Proteins