The Journal of Membrane Biology

, Volume 148, Issue 3, pp 211–222 | Cite as

Excitation-calcium release uncoupling in aged single human skeletal muscle fibers

  • O. Delbono
  • K. S. O'Rourke
  • W. H. Ettinger


The biological mechanisms underlying decline in muscle power and fatigue with age are not completely understood. The contribution of alterations in the excitation-calcium release coupling in single muscle fibers was explored in this work. Single muscle fibers were voltage-clamped using the double Vaseline gap technique. The samples were obtained by needle biopsy of the vastus lateralis (quadriceps) from 9 young (25–35 years; 25.9 ± 9.1; 5 female and 4 male) and 11 old subjects (65–75 years; 70.5 ± 2.3; 6 f, 5 m). Data were obtained from 36 and 39 fibers from young and old subjects, respectively. Subjects included in this study had similar physical activity. Denervated and slow-twitch muscle fibers were excluded from this study. A significant reduction of maximum charge movement (Qmax) and DHP-sensitive Ca current were recorded in muscle fibers from the 65–75 group. Qmax values were 7.6 ± 0.9 and 3.2 ± 0.3 nC/μF for young and old muscle fibers, respectively (P < 0.01). No evidences of charge inactivation or interconversion (charge 1 to charge 2) were found. The peak Ca current was (−)4.7 ± 0.08 and (−)2.15 ± 0.11 μA/μF for young and old fibers, respectively (P < 0.01). The peak calcium transient studied with mag-fura-2 (400 μm) was 6.3 ± 0.4 μm and 4.2 ± 0.3 μm for young and old muscle fibers, respectively. Caffeine (0.5 mm) induced potentiation of the peak calcium transient in both groups. The decrease in the voltage-/ Ca-dependent Ca release ratio in old fibers (0.18 ± 0.02) compared to young fibers (0.47 ± 0.03) (P < 0.01), was recorded in the absence of sarcoplasmic reticulum calcium depletion. These data support a significant reduction of the amount of Ca available for triggering mechanical responses in aged skeletal muscle and, the reduction of Ca release is due to DHPR-ryanodine receptor uncoupling in fast-twitch fibers. These alterations can account, at least partially for the skeletal muscle function impairment associated with aging.

Key words

Dihydropyridine receptor Muscle contraction Calcium release Voltage-clamp Calcium channels Muscle weakness 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anderson, K., Cohn, A.H., Meissner, G. 1994. High-affinity [3H]PN200–110 and [3H]Ryanodine binding to rabbit and frog skeletal muscle. Am. J. Physiol. 266:C462-C466Google Scholar
  2. Bergström, J. 1962. Muscle electrolytes in man. Scand. J. Clin. Invest. Suppl. 68:1–110Google Scholar
  3. Booth, F.W., Weeden, S., Tseng, H. 1993. Effect of aging on human skeletal muscle and motor function. Med. Sci. Sports Ex. 26(5):556–560Google Scholar
  4. Brooks, S.V., Faulkner, J.A. 1988. Contractile properties of skeletal muscles from young, adult and aged mice. J. Physiol. 404:71–82Google Scholar
  5. Brooks, S.V., Faulkner, J.A. 1991. Maximum and sustained power of extensor digitorum longus muscles from young, adult and old mice. J. Gerontol. 46:B28–33Google Scholar
  6. Brooks, S.V., Faulkner, J.A. 1993a. Injury to skeletal muscle fibers during contractions: conditions of occurrence and prevention. Phys. Ther. 73:911–921Google Scholar
  7. Brooks, S.V., Faulkner, J.A. 1993b. Skeletal muscle weakness in old age: underlying mechanisms. Med. Sci. Sports Ex. 26(4):432–439Google Scholar
  8. Brooks, S.V., Faulkner, J.A. 1994a. Skeletal muscle weakness in old age: underlying mechanisms. Med. Sci. Sports Exerc. 26:432Google Scholar
  9. Brooks, S.V., Faulkner, J.A. 1994b. Isometric, shortening, and lengthening contractions of muscle fiber segments from adult and old mice. Am. J. Physiol. 267:C507-C513Google Scholar
  10. Brum, G., Ríos, E., Stefani, E. 1988. Effects of extracellular calcium on the calcium movements of excitation contraction coupling in skeletal muscle fibers. J. Physiol. 398:441–473Google Scholar
  11. Campbell, M.J., McComas, A.J., Petito, F. 1973. Physiological changes in ageing muscles. J. Neurol. Neurosurg. Psychiatry 36:74–182Google Scholar
  12. Caputo, C., Bolaños, P. 1989. Effect of D-600 on intramembrane charge movement of polarized and depolarized frog muscle fibers. J. Gen. Physiol. 94:43–64Google Scholar
  13. Carlson, B.M., Faulkner, J.A. 1988. Reinnervation of long-term denervated rat muscle freely grafted into an innervated limb. Exp. Neurol. Exp. Neurol. 102:50Google Scholar
  14. Caroni, P., Schneider, C. 1994. Signaling by insulin-like growth factors in paralyzed skeletal muscle: rapid induction of IGF1 expression in muscle fibers and prevention of interstitial cell proliferation by IGF-BP5 and IGF-BP4. J. Neurosci. 14(5):3378–3388Google Scholar
  15. Cavalié, A., Berninger, B., Haas, C.A., Garcia, D.E., Lindholm, D., Lux, H.D. 1994. Constitative upregulation of calcium channel currents in rat phaeochromocytoma cells: role of c-fos and c-jun. J. Physiol. 479:11–27Google Scholar
  16. Coggan, A.R., Spina, R.J., King, D.S., Rogers, M.A., Brown Nemeth, P.M., Holloszy, J.O. 1992. Histochemical and enzymatic comparison of the gastrocnemius muscle of young and elderly men and women. J. Gerontol. 47:B71-B76Google Scholar
  17. De Coster, W., De Reuck, J., Sieben, G., Vander Eecken, H. 1981. Early ultrastructural changes in aging rat gastrocnemius muscle: a stereologic study. Muscle Nerve 4:111–116Google Scholar
  18. Delbono, O. 1992. Calcium current activation and charge movement in denervated mammalian skeletal muscle fibres. J. Physiol. 451:187–203Google Scholar
  19. Delbono, O. 1995. Ca2+ modulation of sarcoplasmic reticulum Ca2+ release in mammalian skeletal muscle fibers. J. Membrane Biol. 146:91–99Google Scholar
  20. Delbono, O., Chu, A. 1995. Ca2+ release channels in denervated skeletal muscles. Exp. Physiol 80:561–574Google Scholar
  21. Delbono, O., García, J., Appel, S.H., Stefani, E. 1991. Calcium current and charge movement of mammalian muscle: Action of Amyotrophic Lateral Sclerosis immunoglobulins. J. Physiol. 444:723–742Google Scholar
  22. Delbono, O., Stefani, E. 1993a. Calcium current inactivation in denervated mammalian skeletal muscle fibres. J. Physiol. 460:173–183Google Scholar
  23. Delbono, O., Stefani, E. 1993b. Calcium transients in mammalian skeletal muscle fibers. J. Physiol. 463:689–707Google Scholar
  24. Dulhunty, A.F., Gage, P.W. 1983. Asymmetrical charge movement in slow- and fast-twitch mammalian muscle fibres in normal and paraplegic rats. J. Physiol. 341:213–231Google Scholar
  25. Dulhunty, A.F., Gage, P.W. 1985. Excitation-contraction coupling and charge movement in denervated rat extensor digitorum longus and soleus muscles. J. Physiol. 358:75–89Google Scholar
  26. Edstrom, L., Larsson, L. 1987. Effects of age on contractile and enzyme-histochemical properties of fast- and slow-twitch single motor units in the rat. J. Physiol 392:129–145Google Scholar
  27. Engel, A., Stonnington, H.H. 1974. Morphological effects of denervation of muscle. Ann. NY Acad. Sci. 228:68–88Google Scholar
  28. Fabiato, A. 1983. Calcium-induced release of calcium from the cardiac sarcoplasmic reticulum. Am. J. Physiol. 245:C1-C14Google Scholar
  29. Fitts, R.H., Troup, J.P., Witzmann, F.A., Holloszy, J.O. 1984. The effect of ageing and exercise on skeletal muscle function. Mech. Ageing Dev. 27:161–172Google Scholar
  30. Francini, F., Stefani, E. 1989. Decay of the slow calcium current in twitch muscle fibers of the frog is influenced by intracellular EGTA. J. Gen. Physiol. 94:953–969Google Scholar
  31. Fujisawa, K. 1975. Some observations on the skeletal musculature of aged rats. Part 2. Fine morphology of diseased muscle fibres. J. Neurol. Sci. 24:447–469Google Scholar
  32. García, J., McKinley, K., Appel, S.H., Stefani, E. 1992. Ca2+ current and charge movement in adult single human skeletal muscle fibres. J. Physiol. 454:183–196Google Scholar
  33. García, J., Schneider, M.F. 1993. Calcium transients and calcium release in rat fast-twitch skeletal muscle fibres. J. Physiol. 463:709–728Google Scholar
  34. Gibson, G.E., Petersen, Ch. 1987. Calcium and the Aging nervous system. Neurobiol. Aging 8:329–343Google Scholar
  35. González, A., Ríos, E. 1993. Perchlorate enhances transmission in skeletal muscle excitation-contraction coupling. J. Gen. Physiol. 102:373–421Google Scholar
  36. Grynkiewicz, G., Poenie, M., Tsien, R.Y. 1985. A new generation of calcium indicators with greatly improved fluorescent properties. J. Biol. Chem. 260:3440–3450Google Scholar
  37. Gutmann, E., Carlson, B.M. 1976. Regeneration and transplantation of muscles in old rats and between young and old rats. Life Sci. 18:109–114Google Scholar
  38. Hamilton, S.L., Mejía Alvarez, R., Fill, M., Hawkes, M.J., Brush, K.L., Schilling, W.P., Stefani, E. 1989. [3H]PN200–110 and [3H]Ryanodine binding and reconstitution of ion channel activity with skeletal muscle membranes. Anal. Biochem. 183:31–41Google Scholar
  39. Heizmann, C.W. 1984. Parvalbumin, an intracellular calcium-binding protein: distribution, properties and possible roles in mammalian cells. Experientia 40:910–921Google Scholar
  40. Hodgkin, A., Nakajima, S. 1972. The effect of diameter on the electrical constants of frog skeletal muscle fibres. J. Physiol. 221:105–120Google Scholar
  41. Jacquemond, V., Kao, J.P.Y., Schneider, M.F. 1991. Voltage-gated and calcium-gated release during depolarization of skeletal muscle fibres. Biophys. J. 60:867–873Google Scholar
  42. Johnson, M.A., Polgar, J., Weightman, D., Appleton, D. 1973. Data on the distribution of fibre types in thirty-six human muscles. An autopsy study. J. Neurol. Sci. 18(1):111–129Google Scholar
  43. Kanda, K., Hashizume, H. 1989. Changes in properties of the medial gastrocnemius motor units in aging. J. Neurophysiol. 61:737–746Google Scholar
  44. Klein, M.G., Simon, B.J., Schneider, M.F. 1990. Effects of caffeine release from the sarcoplasmic reticulum in frog skeletal muscle fibres. J. Physiol. 425:599–626Google Scholar
  45. Kotsias, B.A., Muchnik, S. 1987. Mechanical and electrical properties of denervated rat skeletal muscles. Exp. Neurol. 97:516–528Google Scholar
  46. Kotsias, B.A., Muchnik, S., Obejero Paz, C.A. 1986. Co2+, low Ca2+, and verapamil reduce mechanical activity in rat skeletal muscles. Am. J. Physiol. 250:C40–46Google Scholar
  47. Kostyuk, P., Pronchuk, N., Savchenko, A., Verkhratsky, A. 1993. Calcium currents in aged rat dorsal root ganglion neurones. J. Physiol. 461:467–483Google Scholar
  48. Landfield, P.W. 1987. “Increased calcium-current” hypothesis of brain aging. Neurobiol. Aging 8:346–347Google Scholar
  49. Larsson, L., Edström, L. 1986. Effect of age on enzyme-hystochemical fibre spectra and contractile properties of fast- and slow-twitch skeletal muscles in the rat. J. Neurol. Sci. 76:69–89Google Scholar
  50. Larsson, L., Salviati, G. 1992. A technique for studies of the contractile apparatus in single human muscle fibre segments obtained by percutaneous biopsy. Acta Physiol. Scand. 146:485–495Google Scholar
  51. Lexell, J., Henriksson-Larsen, K.B., Winglad, B., Sjostrom, M. 1983. Distribution of different fiber types in human skeletal muscles: effects of aging studied in whole muscle cross sections. Muscle and Nerve 6(8):588–595Google Scholar
  52. Ma, J., Gutierrez, L.M., Hosey, M.M., Hosey, E. 1992. Dihidropyridine-sensitive skeletal muscle Ca channels in polarized planar bilayers. 3. Effects of phosphorylation by protein kinase C. Biophys. J. 63:639–647Google Scholar
  53. Melzer, W., Ríos, E., Schneider, M.F. 1986. The removal of myoplasmic free calcium following calcium release in frog skeletal muscle. J. Physiol. 372:261–292Google Scholar
  54. Melzer, W., Ríos, E., Schneider, M.F. 1987. A general procedure for determining calcium release in skeletal muscle fibers. Biophys. J. 51:849–863Google Scholar
  55. Obejero Paz, C.A., Delbono, O., Muchnik, S. 1986. Effects of actinomycin D on contractile properties of denervated rat skeletal muscle. Exp. Neurol 94:509–518Google Scholar
  56. O'Rourke, K.S., Blaivas, M., Ike, R.W. 1994. Utility of needle biopsy in a university rheumatology practice. J. Rheumatol. 21:413–424Google Scholar
  57. Phillips, S.K., Bruce, S.A., Woledge, R.C. 1991. In mice, the muscle weakness due to age is absent during stretching. J. Physiol. 437:63–70Google Scholar
  58. Phillips, S.K., Wiseman, R.W., Woledge, R.C., Kishmerick, M.J. 1993. Neither changes in phosphorus metabolite levels nor myosin isoforms can explain the weakness in aged mouse muscle. J. Physiol. 463:157–167Google Scholar
  59. Pitler, T.A., Landfield, P.W. 1990. Aging-related prolongation of calcium spike duration in rat hippocampal slice neurons. Brain Res. 508:1–6Google Scholar
  60. Reynolds, J.N., and P.L. Carlen 1989. Diminished calcium currents in aged hippocampal dentate gyrus granule neurones. Brain Res. 479:384–390Google Scholar
  61. Saltin, B., Gollnick, P.D. 1983. Skeletal muscle adaptability: significance for metabolism and performance. In: Handbook of Physiology. Skeletal Muscle. American Physiological Society, editor, p. 572. Bethesda, MarylandGoogle Scholar
  62. Sculptoreanu, A., Scheuer, T., Catterall, W.A. 1993. Voltage-dependent potentiation of L-type Ca2+ channels due to phosphorylation by cAMP-dependent protein kinase. Science 364:240–243Google Scholar
  63. Smith, D.O. 1984. Acetylcholine storage, release and leakage at the neuromuscular junction of mature and aged rats. J. Physiol. 347:161–231Google Scholar
  64. Tanabe, T., Beam, K.G., Powell, J.A., Numa, S. 1988. Restoration of excitation-contraction coupling and slow calcium current in dysgenic muscle by dihydropyridine receptor complementary DNA. Nature 336:134–139Google Scholar
  65. Tanabe, T., Takeshima, H., Mikami, A., Flockerzi, V., Takahashi, H., Kangawa, K., Kojima, H., Matsuo, Hirose, T., Numa, S. 1987. Primary structure of the receptor for calcium channel blockers from skeletal muscle. Nature 328:313–318Google Scholar
  66. Thibault, O., Porter, N.M., Landfield, P.W. 1993. Low Ba2+ and Ca2+ induce a sustained high probability of repolarization openings of L-type Ca2+ channels in hippocampal neurons: physiological implications. Proc. Natl. Acad. Sci. USA 90:11792–11796Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1995

Authors and Affiliations

  • O. Delbono
    • 1
  • K. S. O'Rourke
    • 2
  • W. H. Ettinger
    • 3
  1. 1.Departments of Physiology and Pharmacology, and Internal Medicine (Gerontology)Bowman Gray School of Medicine, Wake Forest UniversityWinston-Salem
  2. 2.Department of Internal Medicine (Rheumatology)Bowman Gray School of Medicine, Wake Forest UniversityWinston-Salem
  3. 3.Departments of Internal Medicine (Gerontology) and Public Health SciencesBowman Gray School of Medicine, Wake Forest University, Medical Center BoulevardWinston-Salem

Personalised recommendations