Advertisement

Journal of Comparative Physiology B

, Volume 181, Issue 1, pp 147–164 | Cite as

Muscle plasticity in hibernating ground squirrels (Spermophilus lateralis) is induced by seasonal, but not low-temperature, mechanisms

  • Megan M. Nowell
  • Hyung Choi
  • Bryan C. RourkeEmail author
Original Paper

Abstract

During hibernation, ground squirrels (Spermophilus lateralis) show unusually altered expression of skeletal muscle myosin heavy-chains. Some muscle groups show transitions from fast to slower myosin isoforms despite atrophy, which are not predicted from other mammalian studies of inactivity. We measure myosin protein and mRNA expression, and the mRNA expression of genes important in atrophy and metabolism in a time-course of muscle plasticity prior to, and during extended hibernation. We also investigate the role of strictly low-temperature processes by comparing torpid individuals at 20 and 4°C. Shifts in myosin isoform expression happen at both temperatures, before the onset of torpor, or within the first month of torpor, in all muscles demonstrating isoform remodeling. Skeletal muscle atrophy is greatly attenuated in this hibernating species, and even may be absent in some muscles. When present, atrophy develops early in hibernation, and does not progress in the final 3 months of torpor. Myostatin mRNA is down-regulated 50–75% in the soleus and diaphragm, two important muscles that are spared of atrophy. The transcription factor FOXO1, which spurs proteolytic degradation of contractile proteins through regulation of the ubiquitin ligase MAFbx, is also generally down-regulated, and may contribute to reduced atrophy. Hypoxia-inducible factor (HIF-1α) mRNA expression was reduced 50% in some muscles, while elevated more than 300% in others. Our collective findings most strongly support early, seasonal, phenotype changes in skeletal muscles which are not uniquely confined to, or prompted by, torpor at 4°C. Such seasonal control of myosin would be a novel mechanism in mammalian skeletal muscle, which otherwise is most susceptible to mechanical loading and limb-activity patterns.

Keywords

Myosin heavy-chains Hibernation Skeletal muscle atrophy Myostatin Hypoxia-inducible factor 

Notes

Acknowledgments

Funding was provided by NIH Minority Biomedical Research Support of Competitive Research 2 S06 GM063119 (BCR). Expert technical assistance was provided by Akino Higa, Justin Neag, Uyen Pham, Yanett Roman, Lauren Sims, and Karine Vu.

Supplementary material

360_2010_505_MOESM1_ESM.pdf (76 kb)
Supplementary material 1 (PDF 75 kb)

References

  1. Abnous K, Storey KB (2007) Regulation of skeletal muscle creatine kinase from a hibernating mammal. Arch Biochem Biophys 467:10–19CrossRefPubMedGoogle Scholar
  2. Adams GR, Caiozzo VJ, Baldwin KM (2003) Skeletal muscle unweighting: spaceflight and ground-based models. J Appl Physiol 95:2185–2201PubMedGoogle Scholar
  3. Allen DL, Unterman TG (2007) Regulation of myostatin expression and myoblast differentiation by FoxO and SMAD transcription factors. Am J Physiol Cell Physiol 292:C188–C199CrossRefPubMedGoogle Scholar
  4. Baldwin KM, Haddad F (2001) Effects of different activity and inactivity paradigms on myosin heavy chain gene expression in striated muscle. J Appl Physiol 90:345–357CrossRefPubMedGoogle Scholar
  5. Barnes BM (1989) Freeze avoidance in a mammal: body temperatures below 0 degree C in an Arctic hibernator. Science 244:1593–1595CrossRefPubMedGoogle Scholar
  6. Bodine SC, Latres E, Baumhueter S, Lai VK, Nunez L, Clarke BA, Poueymirou WT, Panaro FJ, Na E, Dharmarajan K, Pan ZQ, Valenzuela DM, DeChiara TM, Stitt TN, Yancopoulos GD, Glass DJ (2001) Identification of ubiquitin ligases required for skeletal muscle atrophy. Science 294:1704–1708CrossRefPubMedGoogle Scholar
  7. Caiozzo VJ, Baker MJ, Baldwin KM (1997) Modulation of myosin isoform expression by mechanical loading: role of stimulation frequency. J Appl Physiol 82:211–218PubMedGoogle Scholar
  8. Chen J, Sun M, Liang B, Xu A, Zhang S, Wu D (2007) Cloning and expression of PDK4, FOXO1A and DYRK1A from the hibernating greater horseshoe bat (Rhinolophus ferrumequinum). Comp Biochem Physiol B Biochem Mol Biol 146:166–171CrossRefPubMedGoogle Scholar
  9. Choi H, Selpides PJ, Nowell MM, Rourke BC (2009) Functional overload in ground squirrel plantaris muscle fails to induce myosin isoform shifts. Am J Physiol Regul Integr Comp Physiol 297:R578–R586PubMedGoogle Scholar
  10. Dapp C, Gassmann M, Hoppeler H, Fluck M (2006) Hypoxia-induced gene activity in disused oxidative muscle. Adv Exp Med Biol 588:171–188CrossRefPubMedGoogle Scholar
  11. Drummond MJ, Fujita S, Takashi A, Dreyer HC, Volpi E, Rasmussen BB (2008) Human muscle gene expression following resistance exercise and blood flow restriction. Med Sci Sports Exerc 40:691–698CrossRefPubMedGoogle Scholar
  12. Fahlman A, Storey JM, Storey KB (2000) Gene up-regulation in heart during mammalian hibernation. Cryobiology 40:332–342CrossRefPubMedGoogle Scholar
  13. Gayan-Ramirez G, Decramer M (2002) Effects of mechanical ventilation on diaphragm function and biology. Eur Respir J 20:1579–1586CrossRefPubMedGoogle Scholar
  14. Giger JM, Haddad F, Qin AX, Zeng M, Baldwin KM (2005) Effect of unloading on type I myosin heavy chain gene regulation in rat soleus muscle. J Appl Physiol 98:1185–1194CrossRefPubMedGoogle Scholar
  15. Glass DJ (2003a) Signalling pathways that mediate skeletal muscle hypertrophy and atrophy. Nat Cell Biol 5:87–90CrossRefPubMedGoogle Scholar
  16. Glass DJ (2003b) Molecular mechanisms modulating muscle mass. Trends Mol Med 9:344–350CrossRefPubMedGoogle Scholar
  17. Gomes MD, Lecker SH, Jagoe RT, Navon A, Goldberg AL (2001) Atrogin-1, a muscle-specific F-box protein highly expressed during muscle atrophy. Proc Natl Acad Sci USA 98:14440–14445CrossRefPubMedGoogle Scholar
  18. Haddad F, Roy RR, Zhong H, Edgerton VR, Baldwin KM (2003a) Atrophy responses to muscle inactivity. I. Cellular markers of protein deficits. J Appl Physiol 95:781–790PubMedGoogle Scholar
  19. Haddad F, Roy RR, Zhong H, Edgerton VR, Baldwin KM (2003b) Atrophy responses to muscle inactivity. II. Molecular markers of protein deficits. J Appl Physiol 95:791–802PubMedGoogle Scholar
  20. Haddad F, Adams GR, Bodell PW, Baldwin KM (2006) Isometric resistance exercise fails to counteract skeletal muscle atrophy processes during the initial stages of unloading. J Appl Physiol 100:433–441CrossRefPubMedGoogle Scholar
  21. Harlow HJ, Lohuis T, Beck TD, Iaizzo PA (2001) Muscle strength in overwintering bears. Nature 409:997CrossRefPubMedGoogle Scholar
  22. Harlow HJ, Lohuis T, Anderson-Sprecher RC, Beck TD (2004) Body surface temperature of hibernating black bears may be related to periodic muscle activity. J Mammol 85:414–419CrossRefGoogle Scholar
  23. Hershey JD, Robbins CT, Nelson OL, Lin DC (2008) Minimal seasonal alterations in the skeletal muscle of captive brown bears. Physiol Biochem Zool 81:138–147CrossRefPubMedGoogle Scholar
  24. Hodgson JA, Roy RR, Higuchi N, Monti RJ, Zhong H, Grossman E, Edgerton VR (2005) Does daily activity level determine muscle phenotype? J Exp Biol 208:3761–3770CrossRefPubMedGoogle Scholar
  25. Hoehn KL, Hudachek SF, Summers SA, Florant GL (2004) Seasonal, tissue-specific regulation of Akt/protein kinase B and glycogen synthase in hibernators. Am J Physiol Regul Integr Comp Physiol 286:R498–R504PubMedGoogle Scholar
  26. Huey KA, Roy RR, Haddad F, Edgerton VR, Baldwin KM (2002) Transcriptional regulation of the type I myosin heavy chain promoter in inactive rat soleus. Am J Physiol Cell Physiol 282:C528–C537PubMedGoogle Scholar
  27. Huey KA, Haddad F, Qin AX, Baldwin KM (2003) Transcriptional regulation of the type I myosin heavy chain gene in denervated rat roleus. Am J Physiol Cell Physiol 284:C738–C748PubMedGoogle Scholar
  28. Hyatt JP, Roy RR, Baldwin KM, Edgerton VR (2003) Nerve activity-independent regulation of skeletal muscle atrophy: role of MyoD and myogenin in satellite cells and myonuclei. Am J Physiol Cell Physiol 285:C1161–C1173PubMedGoogle Scholar
  29. Hyatt JP, Roy RR, Baldwin KM, Wernig A, Edgerton VR (2006) Activity-unrelated neural control of myogenic factors in a slow muscle. Muscle Nerve 33:49–60CrossRefPubMedGoogle Scholar
  30. Ji M, Zhang Q, Ye J, Wang X, Yang W, Zhu D (2008) Myostatin induces p300 degradation to silence cyclin D1 expression through the PI3K/PTEN/Akt pathway. Cell Signal 20:1452–1458CrossRefPubMedGoogle Scholar
  31. Johnston IA, Temple GK (2002) Thermal plasticity of skeletal muscle phenotype in ectothermic vertebrates and its significance for locomotory behaviour. J Exp Biol 205:2305–2322PubMedGoogle Scholar
  32. Joulia-Ekaza D, Cabello G (2006) Myostatin regulation of muscle development: molecular basis, natural mutations, physiopathological aspects. Exp Cell Res 312:2401–2414CrossRefPubMedGoogle Scholar
  33. Kandarian SC, Jackman RW (2006) Intracellular signaling during skeletal muscle atrophy. Muscle Nerve 33:155–165CrossRefPubMedGoogle Scholar
  34. Kumar S, Nei M, Dudley J, Tamura K (2008) MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Brief Bioinform 9:299–306CrossRefPubMedGoogle Scholar
  35. Latres E, Amini AR, Amini AA, Griffiths J, Martin FJ, Wei Y, Lin HC, Yancopoulos GD, Glass DJ (2005) Insulin-like growth factor-1 (IGF-1) inversely regulates atrophy-induced genes via the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway. J Biol Chem 280:2737–2744CrossRefPubMedGoogle Scholar
  36. Lecker SH, Solomon V, Mitch WE, Goldberg AL (1999) Muscle protein breakdown and the critical role of the ubiquitin-proteasome pathway in normal and disease states. J Nutr 129:227S–237SPubMedGoogle Scholar
  37. Lee K, Park JY, Yoo W, Gwag T, Lee L-W, Byun M-W, Choi I (2008) Overcoming muscle atrophy in a hibernating mammal despite prolonged disuse in dormancy: proteomic and molecular assessment. J Cell Biochem 104:642–656CrossRefPubMedGoogle Scholar
  38. Li ZB, Kollias HD, Wagner KR (2008) Myostatin directly regulates skeletal muscle fibrosis. J Biol Chem 283:19371–19378CrossRefPubMedGoogle Scholar
  39. Lohuis TD, Harlow HJ, Beck TD, Iaizzo PA (2007) Hibernating bears conserve muscle strength and maintain fatigue resistance. Physiol Biochem Zool 80:257–269CrossRefPubMedGoogle Scholar
  40. Mammucari C, Schiaffino S, Sandri M (2008) Downstream of Akt: FoxO3 and mTOR in the regulation of autophagy in skeletal muscle. Autophagy 4:524–526PubMedGoogle Scholar
  41. Martin CI, Johnston IA (2005) The role of myostatin and the calcineurin-signalling pathway in regulating muscle mass in response to exercise training in the rainbow trout Oncorhynchus mykiss Walbaum. J Exp Biol 208:2083–2090CrossRefPubMedGoogle Scholar
  42. Mason S, Johnson RS (2007) The role of HIF-1 in hypoxic response in the skeletal muscle. Adv Exp Med Biol 618:229–244CrossRefPubMedGoogle Scholar
  43. Matsakas A, Diel P (2005) The growth factor myostatin, a key regulator in skeletal muscle growth and homeostasis. Int J Sports Med 26:83–89CrossRefPubMedGoogle Scholar
  44. McCall GE, Allen DL, Haddad F, Baldwin KM (2003) Transcriptional regulation of IGF-I expression in skeletal muscle. Am J Physiol Cell Physiol 285:C831–C839PubMedGoogle Scholar
  45. Milsom WK, Reid WD (1995) Pulmonary mechanics of hibernating squirrels (Spermophilus lateralis). Respir Physiol 101:311–320CrossRefPubMedGoogle Scholar
  46. Morin P Jr, Storey KB (2005) Cloning and expression of hypoxia-inducible factor 1alpha from the hibernating ground squirrel, Spermophilus tridecemlineatus. Biochim Biophys Acta 1729:32–40PubMedGoogle Scholar
  47. Morin P Jr, Storey KB (2006) Evidence for a reduced transcriptional state during hibernation in ground squirrels. Cryobiology 53:310–318CrossRefPubMedGoogle Scholar
  48. Morin P Jr, Dubuc A, Storey KB (2008) Differential expression of microRNA species in organs of hibernating ground squirrels: A role in translational suppression during torpor. Biochim Biophys Acta Gene Regul Mech 1779:628–633Google Scholar
  49. Nelson OL, Robbins CT, Wu Y, Granzier H (2008) Titin isoform switching is a major cardiac adaptive response in hibernating grizzly bears. Am J Physiol Heart Circ Physiol 295:H366–H371CrossRefPubMedGoogle Scholar
  50. Ni YG, Wang N, Cao DJ, Sachan N, Morris DJ, Gerard RD, Kuro OM, Rothermel BA, Hill JA (2007) FoxO transcription factors activate Akt and attenuate insulin signaling in heart by inhibiting protein phosphatases. Proc Natl Acad Sci USA 104:20517–20522CrossRefPubMedGoogle Scholar
  51. Pallafacchina G, Calabria E, Serrano AL, Kalhovde JM, Schiaffino S (2002) A protein kinase B-dependent and rapamycin-sensitive pathway controls skeletal muscle growth but not fiber type specification. PNAS 99:9213–9218CrossRefPubMedGoogle Scholar
  52. Reid WD, Ng A, Wilton R, Milsom WK (1995) Characteristics of diaphragm muscle fibre types in hibernating squirrels. Respir Physiol 101:301–309CrossRefPubMedGoogle Scholar
  53. Rourke BC (2008) Myosin isoform dynamics in muscles of hibernating mammal species: lessons and opportunities. In: Lovegrove BG, McKechnie AE (eds) Hypometabolism in animals: hibernation, torpor and cryobiology. University of ZwaZulu-Natal, Pitermaritzburg, pp 57–64Google Scholar
  54. Rourke BC, Qin A, Haddad F, Baldwin KM, Caiozzo VJ (2004a) Cloning and sequencing of myosin heavy chain isoform cDNAs in golden-mantled ground squirrels: effects of hibernation on mRNA expression. J Appl Physiol 97:1985–1991CrossRefPubMedGoogle Scholar
  55. Rourke BC, Yokoyama Y, Milsom WK, Caiozzo VJ (2004b) Myosin Isoform Expression and MAFbx mRNA Levels in Hibernating Golden-Mantled Ground Squirrels (Spermophilus lateralis). Physiol Biochem Zool 77:582–593CrossRefPubMedGoogle Scholar
  56. Rourke BC, Cotton CJ, Harlow HJ, Caiozzo VJ (2006) Maintenance of slow type I myosin protein and mRNA expression in overwintering prairie dogs (Cynomys leucurus and ludovicianus) and black bears (Ursus americanus). J Comp Physiol [B] 176:709–720Google Scholar
  57. Rourke BC, Selpides PJ, Choi H, Roman Y, Jimenez J (2008) Lack of skeletal muscle atrophy and myosin isoform changes in a rodent hibernator, Spermophilus lateralis, subjected to activity restriction. FASEB J 754:724Google Scholar
  58. Roy RR, Eldridge L, Baldwin KM, Edgerton VR (1996) Neural influence on slow muscle properties: inactivity with and without cross-reinnervation. Muscle Nerve 19:707–714CrossRefPubMedGoogle Scholar
  59. Roy RR, Talmadge RJ, Fox K, Lee M, Ishihara A, Edgerton VR (1997) Modulation of MHC isoforms in functionally overloaded and exercised rat plantaris fibers. J Appl Physiol 83:280–290PubMedGoogle Scholar
  60. Roy RR, Pierotti DJ, Garfinkel A, Zhong H, Baldwin KM, Edgerton VR (2008) Persistence of motor unit and muscle fiber types in the presence of inactivity. J Exp Biol 211:1041–1049CrossRefPubMedGoogle Scholar
  61. Sacheck JM, Ohtsuka A, McLary SC, Goldberg AL (2004) IGF-I stimulates muscle growth by suppressing protein breakdown and expression of atrophy-related ubiquitin ligases, atrogin-1 and MuRF1. Am J Physiol Endocrinol Metab 287:E591–E601CrossRefPubMedGoogle Scholar
  62. Sandri M, Sandri C, Gilbert A, Skurk C, Calabria E, Picard A, Walsh K, Schiaffino S, Lecker SH, Goldberg AL (2004) Foxo transcription factors induce the atrophy-related ubiquitin ligase atrogin-1 and cause skeletal muscle atrophy. Cell 117:399–412CrossRefPubMedGoogle Scholar
  63. Sassoon CS, Caiozzo VJ, Manka A, Sieck GC (2002) Altered diaphragm contractile properties with controlled mechanical ventilation. J Appl Physiol 92:2585–2595PubMedGoogle Scholar
  64. Short KR, Vittone JL, Bigelow ML, Proctor DN, Coenen-Schimke JM, Rys P, Nair KS (2005) Changes in myosin heavy chain mRNA and protein expression in human skeletal muscle with age and endurance exercise training. J Appl Physiol 99:95–102CrossRefPubMedGoogle Scholar
  65. Solomon V, Baracos V, Sarraf P, Goldberg AL (1998) Rates of ubiquitin conjugation increase when muscles atrophy, largely through activation of the N-end rule pathway. Proc Natl Acad Sci USA 95:12602–12607CrossRefPubMedGoogle Scholar
  66. Southgate RJ, Neill B, Prelovsek O, El-Osta A, Kamei Y, Miura S, Ezaki O, McLoughlin TJ, Zhang W, Unterman TG, Febbraio MA (2007) FOXO1 regulates the expression of 4E-BP1 and inhibits mTOR signaling in mammalian skeletal muscle. J Biol Chem 282:21176–21186CrossRefPubMedGoogle Scholar
  67. Steffen JM, Koebel DA, Musacchia XJ, Milsom WK (1991) Morphometric and metabolic indices of disuse in muscles of hibernating ground squirrels. Comp Biochem Physiol B 99:815–819CrossRefPubMedGoogle Scholar
  68. Stitt TN, Drujan D, Clarke BA, Panaro F, Timofeyva Y, Kline WO, Gonzalez M, Yancopoulos GD, Glass DJ (2004) The IGF-1/PI3K/Akt pathway prevents expression of muscle atrophy-induced ubiquitin ligases by inhibiting FOXO transcription factors. Mol Cell 14:395–403CrossRefPubMedGoogle Scholar
  69. Swanson DL, Sabirzhanov B, Vandezande A, Clark TG (2009) Seasonal variation of myostatin gene expression in pectoralis muscle of house sparrows (Passer domesticus) is consistent with a role in regulating thermogenic capacity and cold tolerance. Physiol Biochem Zool 82:121–128CrossRefPubMedGoogle Scholar
  70. Talmadge RJ (2000) Myosin heavy chain isoform expression following reduced neuromuscular activity: potential regulatory mechanisms. Muscle Nerve 23:661–679CrossRefPubMedGoogle Scholar
  71. Talmadge RJ, Roy RR (1993) Electrophoretic separation of rat skeletal muscle myosin heavy-chain isoforms. J Appl Physiol 75:2337–2340PubMedGoogle Scholar
  72. Talmadge RJ, Garcia ND, Roy RR, Edgerton VR (2004) Myosin heavy chain isoform mRNA and protein levels after long-term paralysis. Biochem Biophys Res Commun 325:296–301CrossRefPubMedGoogle Scholar
  73. Templeton GH, Sweeney HL, Timson BF, Padalino M, Dudenhoeffer GA (1988) Changes in fiber composition of soleus muscle during rat hindlimb suspension. J Appl Physiol 65:1191–1195PubMedGoogle Scholar
  74. Thomason DB, Herrick RE, Baldwin KM (1987a) Activity influences on soleus muscle myosin during rodent hindlimb suspension. J Appl Physiol 63:138–144PubMedGoogle Scholar
  75. Thomason DB, Herrick RE, Surdyka D, Baldwin KM (1987b) Time course of soleus muscle myosin expression during hindlimb suspension and recovery. J Appl Physiol 63:130–137PubMedGoogle Scholar
  76. Tinker DB, Harlow HJ, Beck TD (1998) Protein use and muscle-fiber changes in free-ranging, hibernating black bears. Physiol Zool 71:414–424CrossRefPubMedGoogle Scholar
  77. van Breukelen F, Carey V (2002) Ubiquitin conjugate dynamics in the gut and liver of hibernating ground squirrels. J Comp Physiol [B] 172:269–273Google Scholar
  78. van Breukelen F, Martin SL (2002) Reversible depression of transcription during hibernation. J Comp Physiol B 172(5):355–361CrossRefPubMedGoogle Scholar
  79. van Breukelen F, Sonenberg N, Martin SL (2004) Seasonal and state-dependent changes of eIF4E and 4E-BP1 during mammalian hibernation: implications for the control of translation during torpor. Am J Physiol Regul Integr Comp Physiol 287:R349–R353PubMedGoogle Scholar
  80. Velickovska V, Lloyd BP, Qureshi S, van Breukelen F (2005) Proteolysis is depressed during torpor in hibernators at the level of the 20S core protease. J Comp Physiol B 175(5):329–335. doi: 10.1007/s00360-005-0489-x CrossRefPubMedGoogle Scholar
  81. Velickovska V, van Breukelen F (2007) Ubiquitylation of proteins in livers of hibernating golden-mantled ground squirrels, Spermophilus lateralis. Cryobiology 55:230–235CrossRefPubMedGoogle Scholar
  82. Wickler SJ, Hoyt DF, van Breukelen F (1991) Disuse atrophy in the hibernating golden-mantled ground squirrel, Spermophilus lateralis. Am J Physiol 261:R1214–R1217PubMedGoogle Scholar
  83. Willoughby DS, Nelson MJ (2002) Myosin heavy-chain mRNA expression after a single session of heavy-resistance exercise. Med Sci Sports Exerc 34:1262–1269CrossRefPubMedGoogle Scholar
  84. Woods AK, Storey KB (2007) Cytosolic phospholipase A2 regulation in the hibernating thirteen-lined ground squirrel. Cell Mol Biol Lett 12:621–632CrossRefPubMedGoogle Scholar
  85. Wright C, Haddad F, Qin AX, Baldwin KM (1997) Analysis of myosin heavy chain mRNA expression by RT-PCR. J Appl Physiol 83:1389–1396PubMedGoogle Scholar
  86. Zhong H, Roy RR, Hodgson JA, Talmadge RJ, Grossman EJ, Edgerton VR (2002) Activity-independent neural influences on cat soleus motor unit phenotypes. Muscle Nerve 26:252–264CrossRefPubMedGoogle Scholar
  87. Zhu E, Sassoon CS, Nelson R, Pham HT, Zhu L, Baker MJ, Caiozzo VJ (2005) Early effects of mechanical ventilation on isotonic contractile properties and MAF-box gene expression in the diaphragm. J Appl Physiol 99:747–756CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Megan M. Nowell
    • 1
  • Hyung Choi
    • 1
  • Bryan C. Rourke
    • 1
    Email author
  1. 1.Department of Biological SciencesCalifornia State UniversityLong BeachUSA

Personalised recommendations