Advertisement

Gene Hunting in Hypoxia and Exercise

  • Kenneth B. Storey
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 588)

Abstract

New technologies in genomics and proteomics are revolutionizing the study of adaptation to environmental stress. These approaches provide a comprehensive overview of the responses of thousands of genes/proteins to stress and enormously expand our view of the molecular and metabolic changes that underlie physiological responses. Several new technologies can help physiological labs to become gene hunters. DNA array screening is particularly effective for two purposes: (1) identifying coordinated responses by functional groups of gene/proteins such as multiple members of a signal transduction cascade or enzymes of a metabolic pathway, and (2) highlighting cell functions that have never before been linked with the stress under consideration. We have shown that heterologous screening of DNA arrays can be a highly effective method of gene hunting for the comparative biochemist provided that it is followed up by species-specific analyses including PCR to quantify transcript levels and Western blotting to analyze protein responses. Recent work in my lab has used cDNA array screening to evaluate responses to low oxygen by multiple hypoxia/anoxia tolerant systems, revealing common gene responses across phylogeny. Analysis of vertebrate facultative anaerobiosis in freshwater turtles reveals an interesting mixture of gene responses, including up-regulation of antioxidant enzymes, protease inhibitors, and proteins of iron metabolism; a few of these are coordinated by the hypoxia inducible factor in other systems but most are not. Array screening is also providing new insights into how exercise stimulates the growth of differentiated muscle cells and studies in our lab are identifying the gene responses associated with “anti-exercise”-gene up-regulation that aids hibernating mammals to maintain their muscle mass despite months of inactivity.

Key Words

anoxia tolerance hibernation cDNA array screening metabolic rate depression 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Andrews MT. Genes controlling the metabolic switch in hibernating mammals. Biochem Soc Trans 32: 1021–1024, 2004.PubMedCrossRefGoogle Scholar
  2. 2.
    Bickler PE, Donohoe PH, and Buck LT. The hypoxic brain: suppressing energy-expensive membrane functions by regulation of receptors and ion channels. In: Molecular Mechanisms of Metabolic Arrest, edited by Storey KB. Oxford: BIOS Scientific, 2001, p. 77–102.Google Scholar
  3. 3.
    Buzadzic B, Spasic M, Saicic ZS, Radojicic R, Petrovic VM, Halliwell B. Antioxidant defenses in the ground squirrel Citellus citellus. 2. The effect of hibernation. Free Radic Biol Med 9: 407–413, 1990.PubMedCrossRefGoogle Scholar
  4. 4.
    Cai Q, and Storey KB. Anoxia-induced gene expression in turtle heart: up-regulation of mitochondrial genes for NADH-ubiquinone oxidoreductase subunit 5 and cytochrome C oxidase subunit 1. Eur J Biochem 241: 83–92, 1996.PubMedCrossRefGoogle Scholar
  5. 5.
    Douglas RM, and Haddad GG. Genetic models in applied physiology: invited review: effect of oxygen deprivation on cell cycle activity: a profile of delay and arrest. J Appl Physiol 94: 2068–2083, 2003.PubMedGoogle Scholar
  6. 6.
    Duh EJ, Yang HS, Suzuma I, Miyagi M, Youngman E, Mori K, Katai M, Yan L, Suzuma K, West K, Davarya S, Tong P, Gehlbach P, Pearlman J, Crabb JW, Aiello LP, Campochiaro PA, and Zack DJ. Pigment epithelium-derived factor suppresses ischemia-induced retinal neovascularization and VEGF-induced migration and growth. Invest Ophthalmol Vis Sci 43: 821–829, 2002.PubMedGoogle Scholar
  7. 7.
    Eddy SF, and Storey KB. Dynamic use of cDNA arrays: heterologous probing for gene discovery and exploration of animal adaptations in stressful environments. In: Cell and Molecular Responses to Stress, edited by Storey KB and Storey JM. Amsterdam: Elsevier 2002, vol. 3, p. 315–325.Google Scholar
  8. 8.
    Eddy SF, and Storey KB. Differential expression of Akt, PPAR-γ and PGC-1 during hibernation in bats. Biochem Cell Biol 81: 269–274, 2003.PubMedCrossRefGoogle Scholar
  9. 9.
    Eddy SF, and Storey KB. Up-regulation of fatty acid-binding proteins during hibernation in the little brown bat, Myotis lucifugus. Biochim Biophys Acta 1676: 63–70, 2004.Google Scholar
  10. 10.
    Eddy SF, McNally JD, and Storey KB. Up-regulation of a thioredoxin peroxidase-like protein, proliferation associated gene, in hibernating bats. Arch Biochem Biophys 435: 101–111, 2005.CrossRefGoogle Scholar
  11. 11.
    Egginton S, Fairney J, and Bratcher J. Differential effects of cold exposure on muscle fibre composition and capillary supply in hibernator and non-hibernator rodents. Exp Physiol 86: 629–639, 2001.PubMedCrossRefGoogle Scholar
  12. 12.
    Fahlman A, Storey JM, and Storey KB. Gene up-regulation in heart during mammalian hibernation. Cryobiology 40: 332–342, 2000.PubMedCrossRefGoogle Scholar
  13. 13.
    Fletcher GL, Hew CL, and Davies PL. Antifreeze proteins of teleost fish. Ann Rev Physiol 63: 359–390, 2001.CrossRefGoogle Scholar
  14. 14.
    Fluck M, and Hoppeler H. Molecular basis of skeletal muscle plasticity-from gene to form and function. Rev Physiol Biochem Pharmacol 146: 159–216, 2003.PubMedGoogle Scholar
  15. 15.
    Fujii J, and Ikeda YAdvances in our understanding of peroxiredoxin, a multifunctional, mammalian redox protein. Redox Rep 7: 123–130, 2002.PubMedCrossRefGoogle Scholar
  16. 16.
    Gettins PGW. Serpin structure, mechanism and function. Chem Rev 102: 4751–4803, 2002.PubMedCrossRefGoogle Scholar
  17. 17.
    Haddad JJ. Oxygen-sensing mechanisms and the regulation of redox-responsive transcription factors in development and pathophysiology. Respir Res. 3: 26, 2002. http://respiratory-research.com/content/3/1/ 126 PubMedCrossRefGoogle Scholar
  18. 18.
    Hentze MW, Muckenthaler MU, and Andrews NC. Balancing acts: molecular control of mammalian iron metabolism. Cell 117: 285–297, 2004.PubMedCrossRefGoogle Scholar
  19. 19.
    Hermes-Lima M, Zenteno-Savin T. Animal response to drastic changes in oxygen availability and physiological oxidative stress. Comp Biochem Physiol C 133: 537–556, 2002.CrossRefGoogle Scholar
  20. 20.
    Hermes-Lima M, Storey JM, Storey KB. Antioxidant defenses and animal adaptation to oxygen availability during environmental stress. In: Cell and Molecular Responses to Stress edited by Storey KB and Storey JM. Amsterdam: Elsevier Press, 2001, vol. 2, p. 263–287.Google Scholar
  21. 21.
    Hittel D, and Storey KB. Differential expression of adipose and heart type fatty acid binding proteins in hibernating ground squirrels. Biochim Biophys Acta 1522: 238–243, 2001.PubMedGoogle Scholar
  22. 22.
    Hittel D, and Storey KB. Differential expression of mitochondria-encoded genes in a hibernating mammal. J Exp Biol 205: 1625–1631, 2002.PubMedGoogle Scholar
  23. 23.
    Hittel D, and Storey KB. The translation state of differentially expressed mRNAs in the hibernating thirteen-lined ground squirrel (Spermophilus tridecemlineatus). Arch Biochem Biophys 401: 244–254, 2002.PubMedCrossRefGoogle Scholar
  24. 24.
    Hochachka PW, Buck LT, Doll CJ, and Land SC. Unifying theory of hypoxia tolerance: molecular/metabolic defense and rescue mechanisms for surviving oxygen lack. Proc Natl Acad Sci USA 93: 9493–9498, 1996.PubMedCrossRefGoogle Scholar
  25. 25.
    Hochachka PW, and Lutz PL. Mechanism, origin and evolution of anoxia tolerance in animals. Comp Biochem Physiol B 130: 435–459, 2001.PubMedCrossRefGoogle Scholar
  26. 26.
    Jackson DC. How a turtle’s shell helps it survive prolonged anoxic acidosis. News Physiol Sci 15: 181–185, 2000.PubMedGoogle Scholar
  27. 27.
    Jin DY, Chae HZ, Rhee SG, Jeang KT. Regulatory role for a novel human thioredoxin peroxidase in NF-κB activation. J Biol Chem 272: 30952–30961, 1997.PubMedCrossRefGoogle Scholar
  28. 28.
    Larade K, and Storey KB. Characterization of a novel gene up-regulated during anoxia exposure in the marine snail Littorina littorea. Gene 283: 145–154, 2002.CrossRefGoogle Scholar
  29. 29.
    Larade K, and Storey KB. Accumulation and translation of ferritin heavy chain transcripts following anoxia exposure in a marine invertebrate. J Exp Biol 207: 1353–1360, 2004.PubMedCrossRefGoogle Scholar
  30. 30.
    Larade K, and Storey KB. Anoxia-induced transcriptional up-regulation of sarp-19: cloning and characterization of a novel EF-hand containing gene expressed in hepatopancreas of Littorina littorea. Biochem Cell Biol 82: 285–293, 2004.PubMedCrossRefGoogle Scholar
  31. 31.
    Lutz PL, and Storey KB. Adaptations to variations in oxygen tension by vertebrates and invertebrates. In: Handbook of Physiology, Section 13: Comparative Physiology, edited by Dantzler WH. Oxford: Oxford University Press, 1997, Vol. 2, pp. 1479–1522.Google Scholar
  32. 32.
    MacDonald JA, and Borman MA. Analyzing biological function with emerging proteomic technologies. Int Cong Ser 1275: 14–21, 2004.CrossRefGoogle Scholar
  33. 33.
    Morano I, Adler K, Agostini B, and Hasselbach W. Expression of myosin heavy and light chains and phosphorylation of myosin light chain in the heart ventricle of the European hamster during hibernation and in summer. J Muscle Res Cell Motil 13: 64–70, 1992.PubMedCrossRefGoogle Scholar
  34. 34.
    Morin P, and Storey KB. Cloning and expression of hypoxia-inducible factor 1α from the hibernating ground squirrel, Spermophilus tridecemlineatus. Biochim Biophys Acta, 1729(1): 32–40, 2005.PubMedGoogle Scholar
  35. 35.
    Petersen SV, Valnickova Z, and Enghild JJ. Pigment-epithelium-derived factor (PEDF) occurs at a physiologically relevant concentration in human blood: purification and characterization. Biochem J 374: 199–206, 2003.PubMedCrossRefGoogle Scholar
  36. 36.
    Powers SK, Kavazis AN, and DeRuisseau KC. Mechanisms of disuse muscle atrophy: role of oxidative stress. Am J Physiol 288: R337–R344, 2005.Google Scholar
  37. 37.
    Prentice HM, Milton SL, Scheurle D, and Lutz PL. Gene transcription of brain voltage-gated potassium channels is reversibly regulated by oxygen supply. Am J Physiol 285: R1317–1321, 2003.Google Scholar
  38. 38.
    Rankinen T, Perusse L, Rauramaa R, Rivera MA, Wolfarth B, and Bouchard C. The human gene map for performance and health-related fitness phenotypes: the 2003 update. Med Sci Sports Exerc 36: 1451–1469, 2004.PubMedCrossRefGoogle Scholar
  39. 39.
    Rhee SG, Chang T-S, Bae YS, Lee S-R, and Kang SW. Cellular regulation by hydrogen peroxide. J Am Soc Nephrol 14: S211–S215, 2003.PubMedCrossRefGoogle Scholar
  40. 40.
    Rodriguez OC, and Cheney RE. Human myosin-Vc is a novel class V myosin expressed in epithelial cells. J Cell Sci 115: 991–1004, 2002.PubMedGoogle Scholar
  41. 41.
    Smith JJ, O’Brien-Ladner AR, Kaiser CR, and Wesselius LJ. Effects of hypoxia and nitric oxide on ferritin content of alveolar cells. J Lab Clin Med 141: 309–317, 2003.PubMedCrossRefGoogle Scholar
  42. 42.
    Steffen JM, Koebel DA, Musacchia XJ, and Milsom WK. Morphometric and metabolic indices of disuse in muscles of hibernating ground squirrels. Comp Biochem Physiol B 99: 815–819, 1991.PubMedCrossRefGoogle Scholar
  43. 43.
    Storey KB. Mammalian hibernation: transcriptional and translational controls. Adv Exp MedBiol 543: 21–38, 2003.Google Scholar
  44. 44.
    Storey KB. Cold, ischemic organ preservation: lessons from natural systems. J Invest Med 52: 315–322, 2004.Google Scholar
  45. 45.
    Storey KB. Gene regulation in physiological stress. Int Cong Ser 1275: 1–13, 2004.CrossRefGoogle Scholar
  46. 46.
    Storey KB. Molecular mechanisms of anoxia tolerance. Int Cong Ser 1275: 47–54, 2004.CrossRefGoogle Scholar
  47. 47.
    Storey KB. Strategies for exploration of freeze responsive gene expression: advances in vertebrate freeze tolerance. Cryobiology 48: 134–145, 2004.PubMedCrossRefGoogle Scholar
  48. 48.
    Storey KB. Hibernating mammals: can natural cryoprotective mechanisms help prolong lifetimes of transplantable organs? In: Extending the Life Span, edited by Sames K, Sethe S. and Stolzing A. Transaction Publishers, NY, 2005.Google Scholar
  49. 49.
    Storey KB, and McMullen DC. Insect cold-hardiness: new advances using gene screening technology. In: Life in the Cold: Evolution, Mechanisms, Adaptation, and Application. Edited by Barnes BM, and Carey HV. Fairbanks: Biological Papers of the University of Alaska, 2004, #27, p. 275–281.Google Scholar
  50. 50.
    Storey KB, and Storey JM. Natural freeze tolerance in ectothermic vertebrates. Ann Rev Physiol 54: 619–637, 1992.CrossRefGoogle Scholar
  51. 51.
    Storey KB, and Storey JM. Metabolic rate depression in animals: transcriptional and translational controls. Biol Rev Camb Philos Soc 79: 207–233, 2004.PubMedCrossRefGoogle Scholar
  52. 52.
    Tacchini L., Fusar Poli D, Bernelli-Zazzera A, and Cairo G. Transferrin receptor gene expression and transferrin-bound iron uptake are increased during postischemic rat liver perfusion. Hepatology 36: 103–111, 2002.PubMedCrossRefGoogle Scholar
  53. 53.
    Wang LCH, and Lee TF. Torpor and hibernation in mammals: metabolic, physiological, and biochemical adaptations. In: Handbook of Physiology: Environmental Physiology, edited by Fregley MJ, and Blatteis CM. New York: Oxford University Press, 1996, sect. 4, vol. 1, p. 507–532.Google Scholar
  54. 54.
    Wenger RH. Cellular adaptation to hypoxia: O2-sensing protein hydroxylases, hypoxia-inducible transcription factors, and O2-regulated gene expression. FASEB J 16: 1151–1162, 2002.PubMedCrossRefGoogle Scholar
  55. 55.
    Willmore WG, English TE, and Storey KB. Mitochondrial gene responses to low oxygen stress in turtle organs. Copeia 2001, 628–637, 2001.CrossRefGoogle Scholar
  56. 56.
    Wittwer M, Billeter R, Hoppeler H, and Fluck M. Regulatory gene expression in skeletal muscle of highly endurance-trained humans. Acta Physiol Scand 180: 217–227, 2004.PubMedCrossRefGoogle Scholar
  57. 57.
    Wood ZA, Schroder E, Harris JR, and Poole LB. Structure, mechanism and regulation of peroxiredoxins. Trends Biochem Sci 28: 32–40, 2003.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2006

Authors and Affiliations

  • Kenneth B. Storey
    • 1
  1. 1.Institute of BiochemistryCarleton UniversityOttawaCanada

Personalised recommendations