Journal of Molecular Histology

, Volume 35, Issue 5, pp 489–499

Early Onset of Lipofuscin Accumulation in Dystrophin-Deficient Skeletal Muscles of DMD Patients and mdx Mice

  • Yoshiko Nakae
  • Peter J. Stoward
  • Tatsuo Kashiyama
  • Masayuki Shono
  • Akiko Akagi
  • Tetsuya Matsuzaki
  • Ikuya Nonaka
Article

Abstract

Lipofuscin, the so-called ageing pigment, is formed by the oxidative degradation of cellular macromolecules by oxygen-derived free radicals and redox-active metal ions. Usually it accumulates in post-mitotic, long-lived cells such as neurons and cardiac muscle cells. In contrast, it is rarely seen in either normal or diseased skeletal muscle fibres. In this paper, we report that lipofuscin accumulates at an early age in both human and murine dystrophic muscles. Autofluorescent lipofuscin granules were localized, using confocal laser scanning microscopy and electron microscopy, in dystrophin-deficient skeletal muscles of X chromosome-linked young Duchenne muscular dystrophy (DMD) patients and of mdx mice at various ages after birth. Age-matched normal controls were studied similarly. Autofluorescent lipofuscin granules were observed in dystrophic biceps brachii muscles of 2–7-year-old DMD patients where degeneration and regeneration of myofibres are active, but they were rarely seen in age-matched normal controls. In normal mice, lipofuscin first appears in diaphragm muscles nearly 20 weeks after birth but in mdx muscles it occurs much earlier, 4 weeks after birth, when the primary degeneration of dystrophin-deficient myofibres is at a peak. Lipofuscin accumulation increases with age in both mdx and normal controls and is always higher in dystrophic muscles than in age-matched normal controls. At the electron microscopical level, it was confirmed that the localisation of autofluorescent granules observed by light microscopy in dystrophin-deficient skeletal muscles coincided with lipofuscin granules in myofibres and myosatellite cells, and in macrophages accumulating around myofibres and in interstitial connective tissue. Our results agree with previous biochemical and histochemical data implying increased oxidative damages in DMD and mdx muscles. They indicate that dystrophin-deficient myofibres are either more susceptible to oxidative stress, or are subjected to higher intra- or extracellular oxidative stress than normal controls, or both.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Beregi E, Regius O, Hüttl T, Göbl Z (1988) Age-related changes in the skeletal muscle cells. Z Gerontol 21: 83-86.Google Scholar
  2. Brenman JE, Chao DS, Xia H, Aldape K, Bredt DS (1995) Nitric oxide synthase complexed with dystrophin and absent from skeletal muscle sarcolemma in Duchenne muscular dystrophy. Cell 82: 743-752.Google Scholar
  3. Bulfield G, Siller WG, Wight PAL, Moore KJ (1984) X chromosome-linked muscular dystrophy (mdx) in the mouse. Proc Natl Acad Sci USA 81: 1189-1192.Google Scholar
  4. Chang W-J, Iannaccone ST, Lau KS, Masters BSS, McCabe TJ, McMillan K, Padre RC, Spencer MJ, Tidball JG, Stull JT (1996) Neuronal nitric oxide synthase and dystrophin-deficient muscular dystrophy. Proc Natl Acad Sci USA 93: 9142-9147.Google Scholar
  5. Chao DS, Silvagno F, Bredt DS (1998) Muscular dystrophy in mdx mice despite lack of neuronal nitric oxide synthase. J Neurochem 71: 784-789.Google Scholar
  6. Crosbie RH, Straub V, Yun H-Y, Lee JC, Rafael JA, Chamberlain JS, Dawson VL, Dawson TM, Campbell KP(1998) Mdx muscle pathology is independent of nNOS perturbation. Hum Mol Genet 7: 823-829.Google Scholar
  7. Darley-Usmar V, Wiseman H, Halliwell B (1995) Nitric oxide and oxygen radicals: A question of balance. FEBS Lett 369: 131-135.Google Scholar
  8. Disatnik M-H, Dhawan J, Yu Y, Beal MF, Whirl MM, Franco AA, Rando TA (1998) Evidence of oxidative stress in mdx mouse muscle: Studies of the pre-necrotic state. J Neurol Sci 161: 77-84.Google Scholar
  9. Disatnik M-H, Chamberlain JS, Rando TA (2000) Dystrophin mutations predict cellular susceptibility to oxidative stress. Muscle Nerve 23: 784-792.Google Scholar
  10. Dupont-Versteegden EE, McCarter RJ (1992) Differential expression of muscular dystrophy in diaphragm versus hindlimb muscles of mdx mice. Muscle Nerve 15: 1105-1110.Google Scholar
  11. Engel AG, Banker BQ (1994) Ultrastructural changes in diseased muscle. In: Engel AG, Franzini-Armstrong C, eds. Myology: Basic and Clinical. Vol. 1, 2nd edn. New York: McGraw-Hill, pp. 889-1017.Google Scholar
  12. Faist V, Koenig J, Hoeger H, Elmadfa I (1998) Mitochondrial oxygen consumption, lipid peroxidation and antioxidant enzyme systems in skeletal muscle of senile dystrophic mice. Pflügers Arch-Eur J Physiol 437: 168-171.Google Scholar
  13. Faist V, König J, Höger H, Elmadfa I (2001) Decreased mitochondrial oxygen consumption and antioxidant enzyme activities in skeletal muscle of dystrophic mice after low-intensity exercise. Ann Nutr Metab 45: 58-66.Google Scholar
  14. Grozdanovic Z, Gosztonyi G, Gossrau R (1996) Nitric oxide synthase I (NOS-I) is deficient in the sarcolemma of striated muscle fibres in patients with Duchenne muscular dystrophy, suggesting an association with dystrophin. Acta Histochem 98: 61-69.Google Scholar
  15. Haycock JW, Neil SM, Jones P, Harris JB, Mantle D (1996) Oxidative damage to muscle protein in Duchenne muscular dystrophy. NeuroReport 8: 357-361.Google Scholar
  16. Hoffman EP, Brown RH, Kunkel LM (1987) Dystrophin: The protein product of the Duchenne muscular dystrophy locus. Cell 51: 919-928.Google Scholar
  17. Ikeda H, Tauchi H, Sato T (1985) Fine structural analysis of lipofuscin in various issues of rats of different ages. Mech Ageing Dev 33: 77-93.Google Scholar
  18. Kameya S, Miyagoe Y, Nonaka I, Ikemoto T, Endo M, Hanaoka K, Nabeshima Y, Takeda S (1999) á 1-syntrophin gene disruption results in the absence of neuronal-type nitric-oxide synthase at the sarco-lemma but does not induce muscle degeneration. J Biol Chem 274: 2193-2200.Google Scholar
  19. Kárpáti G, Carpenter S, Wolfe LS (1988) Clinical and experimental studies on lipofuscin in skeletal muscle fibres. In: Zs-Nagy I, ed. Lipofuscin-1987: State of the Art, Budapest: Akadémiai Kiadò and Amsterdam: Elsevier Science Publishers, pp. 227-244.Google Scholar
  20. Kelley EE, Wagner BA, Buettner GR, Burns CP (1999) Nitric oxide inhibits iron-induced lipid peroxidation in HL-60 cells. Arch Biochem Biophys 370: 97-104.Google Scholar
  21. Laule S, Bornemann A (2001) Ultrastructural findings at the satellite cell-myofibre border in normal and diseased human muscle biopsy specimens. Acta Neuropathol 101: 435-439.Google Scholar
  22. Louboutin JP, Fichter-Gagnepain V, Thaon E, Fardeau M (1993) Morphometric analysis of mdx diaphragm muscle fibres. Comparison with hindlimb muscles. Neuromuscul Disord 3: 463-469.Google Scholar
  23. Louboutin J-P, Rouger K, Tinsley JM, Halldorson J, Wilson JM (2001) iNOS expression in dystrophinopathies can be reduced by somatic gene transfer of dystrophin or utrophin. Mol Med 7: 355-364.Google Scholar
  24. Mastaglia FL, Papadimitriou JM, Kakulas BA (1970) Regeneration of muscle in Duchenne muscular dystrophy: An electron microscope study. J Neurol Sci 11: 425-444.Google Scholar
  25. McKelvie P, Friling R, Davey K, Kowal L (1999) Changes as the result of ageing in extraocular muscles: A post-mortem study. Aust NZ J Ophthalmol 27: 420-425.Google Scholar
  26. Murphy ME, Kehrer JP (1989) Oxidative stress and muscular dystrophy. Chem Biol Interact 69: 101-173.Google Scholar
  27. Nakae Y, Stoward PJ, Shono M, Matsuzaki T (1999) Localisation and quantification of dehydrogenase activities in single muscle fibres of mdx gastrocnemius. Histochem Cell Biol 112: 427-436.Google Scholar
  28. Nakae Y, Stoward PJ, Shono M, Matsuzaki T (2001) Most apoptotic cells in mdx diaphragm muscle contain accumulated lipofuscin. Histochem Cell Biol 115: 205-214.Google Scholar
  29. Petrof BJ, Shrager JB, Stedman HH, Kelly AM, Sweeney HL (1993) Dystrophin protects the sarcolemma from stresses devel-oped during muscle contraction. Proc Natl Acad Sci USA 90: 3710-3714.Google Scholar
  30. Ragusa RJ, Chow CK, Porter JD (1997) Oxidative stress as a potential pathogenic mechanism in an animal model of Duchenne muscular dystrophy. Neuromuscul Disord 7: 379-386.Google Scholar
  31. Rando TA, Disatnik M-H, Yu Y, Franco A (1998) Muscle cells from mdx mice have an increased susceptibility to oxidative stress. Neuromuscul Disord 8: 14-21.Google Scholar
  32. Roth SM, Martel GF, Ivey FM, Lemmer JT, Metter EJ, Hurley BF, Rogers MA (2000) Skeletal muscle satellite cell populations in healthy young and older men and women. Anat Rec 260: 351-358.Google Scholar
  33. Schnell SA, Staines WA, Wessendorf MW (1999) Reduction of lipofuscin-like autofluorescence in fluorescently labeled tissue. J Histochem Cytochem 47: 719-730.Google Scholar
  34. Schultz E (1976) Fine structure of satellite cells in growing skeletal muscle. Am J Anat 147: 49-70.Google Scholar
  35. Stedman HH, Sweeney HL, Shrager JB, Maguire HC, Panettieri RA, Petrof B, Narusawa M, Leferovich JM, Sladky JT, Kelly AM (1991) The mdx mouse diaphragm reproduces the degenerative changes of Duchenne muscular dystrophy. Nature 352: 536-539.Google Scholar
  36. Terman A, Brunk UT (1998a) Lipofuscin: Mechanisms of formation and increase with age. APMIS 106: 265-276.Google Scholar
  37. Terman A, Brunk UT (1998b) On the degradability and exocytosis of ceroid/lipofuscin in cultured rat cardiac myocytes. Mech Ageing Dev 100: 145-156.Google Scholar
  38. Wehling M, Stull JT, McCabe TJ, Tidball JG (1998) Sparing of mdx extraocular muscles from dystrophic pathology is not attributable to normalized concentration or distribution of neuronal nitric oxide synthase. Neuromuscul Disord 8: 22-29.Google Scholar
  39. Wehling M, Spencer MJ, Tidball JG (2001) Anitric oxide synthase trans-gene ameliorates muscular dystrophy in mdx mice. J Cell Biol 155: 123-131.Google Scholar
  40. Wink DA, Hanbauer I, Krishna MC, DeGraff W, Gamson J, Mitchell JB (1993) Nitric oxide protects against cellular damage and cyto-toxicity from reactive oxygen species. Proc Natl Acad Sci USA 90: 9813-9817.Google Scholar
  41. Yin D (1996) Biochemical basis of lipofuscin, ceroid, and age pigment-like fluorophores. Free Rad Biol Med 21: 871-888.Google Scholar
  42. Zhuang W, Eby JC, Cheong M, Mohapatra PK, Bredt DS, Disatnik M-H, Rando TA (2001) The susceptibility of muscle cells to oxidative stress is independent of nitric oxide synthase expression. Muscle Nerve 24: 502-511.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Yoshiko Nakae
    • 1
  • Peter J. Stoward
    • 2
  • Tatsuo Kashiyama
    • 3
  • Masayuki Shono
    • 3
  • Akiko Akagi
    • 4
  • Tetsuya Matsuzaki
    • 5
  • Ikuya Nonaka
    • 6
  1. 1.Department of Oral Anatomy 1Tokushima University School of DentistryTokushimaJapan
  2. 2.School of Life SciencesUniversity of DundeeDundeeUK
  3. 3.General Laboratory for Medical ResearchTokushima University School of MedicineTokushimaJapan
  4. 4.Matsui Hospital/Japan Neurological InstituteKagawaJapan
  5. 5.Central Institute for Experimental AnimalsKawasakiJapan
  6. 6.National Centre of Neurology and PsychiatryKodairaJapan

Personalised recommendations