Acta Neuropathologica

, Volume 80, Issue 3, pp 239–250 | Cite as

Heterogeneity of dystrophin expression in patients with Duchenne and Becker muscular dystrophy

  • L. V. B. Nicholson
  • M. A. Johnson
  • D. Gardner-Medwin
  • S. Bhattacharya
  • J. B. Harris
Regular Papers

Summary

This report documents the results of an integrated biochemical and immunocytochemical investigation into the expression of dystrophin (the protein product of the Duchenne muscular dystrophy gene) in muscle biopsies from 226 patients. It is the first study in which dystrophin has been analysed on blots and on tissue sections in such a large number of patients using the same (monoclonal) antibody. The 140 patients with Xp21 muscular dystrophy who were included in this study represent a continuous spectrum of disease severity and this range was reflected in the heterogeneity of dystrophin expression which was observed with respect to abundance, size and the pattern of tissue localisation. Approximately 40% of biopsies obtained from patients diagnosed as having Duchenne muscular, dystrophy (DMD) contained isolated clearly positive fibres and a further 20% had very weak labelling on a large number of fibres. Biopsies from patients with Becker muscular dystrophy (BMD) showed labelling patterns which varied from weak labelling on the majority of fibres to clear labelling on all fibres. Typically, however, there was inter-and intra-fibre variation in labelling intensity. Approximately 85% of the 52 BMD and 54 DMD patients who had unequivocal labelling on blots demonstrated a protein of abnormal size. The remaining 15% had a protein of normal size but reduced abundance. Overall, the estimated abundance of dystrophin correlated well with clinical assessments of the disease severity expressed in patients: We conclude that dystrophin analysis is an essential and dependable technique for the differential diagnosis of patients with Xp21 muscular dystrophy.

Key words

Dystrophin Duchenne muscular dystrophy Becker muscular dystrophy Muscle proteins 

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References

  1. 1.
    Arahata K, Ishiura S, Ishiguro T, Tsukahara T, Suhara Y, Eguchi C, Ishihara T, Nonaka I, Ozawa E, Sugita H (1988) Immunostaining of skeletal and cardiac muscle surface membrane with antibody against Duchenne muscular dystrophy peptide. Nature 333:861–863Google Scholar
  2. 2.
    Arahata K, Ishiura S, Tsukahara T, Sugita H (1989) Dystrophin digest. Nature 337: 606 (Addendum, to ref. 1)Google Scholar
  3. 3.
    Arahata K, Hoffman EP, Kunkel LM, Ishiura S, Tsukahara T, Ishihara T, Sunohara N, Nonaka I, Ozawa E, Sugita H (1989) Dystrophin diagnosis: comparison of dystrophin abnormalities by immunofluorescence and immunoblot analyses. Proc Natl Acad Sci USA 86:7154–7148Google Scholar
  4. 4.
    Arahata K, Ishihara T, Kamakura K, Tsukahara T, Ishiura S, Baba C, Matsumoto T, Nonaka I, Sugita H (1989) Mosaic expression of dystrophin in symptomatic carriers of Duchenne's muscular dystrophy. N Engl J Med 320: 138–142Google Scholar
  5. 5.
    Baumbach LL, Chamberlain JS, Ward PA, Farwell NJ, Caskey CT (1989) Molecular and clinical correlations of deletions leading to Duchenne and Becker muscular dystrophies. Neurology 39:465–474Google Scholar
  6. 6.
    Becker PE Kiener F (1955) Eine neue X-chromosomale Muskeldystrophie. Arch Psychiatr Z Neurol 193:427–448Google Scholar
  7. 7.
    Bonilla E, Samitt CE, Miranda AF, Hays AP, Salviati G, DiMauro S, Kunkel LM, Hoffman EP, Rowland LP (1988) Duchenne muscular dystrophy: deficiency of dystrophin at the muscular cell surface. Cell 54:447–452Google Scholar
  8. 8.
    Bonilla S, Schmidt B, Samitt CE, Miranda AF, Hays AP, De Oliveira ABS, Chang HW, Servidei S, Ricci E, Younger DS, DiMauro S (1988) Normal and dystrophin-deficient muscle fibers in carriers of the gene for Duchenne. Am J Pathol 133:440–445Google Scholar
  9. 9.
    Bradley WG, Jones MZ, Mussini JM, Fawcett PRW (1978) Becker-type muscular dystrophy. Muscle Nerve 1:111–132Google Scholar
  10. 10.
    Campbell KP, Kahl SD (1989) Association of dystrophin and an integral membrane glycoprotein. Nature 338:259–262Google Scholar
  11. 11.
    Dickson G, Pizzey JA, Elsom VE, Love D, Davies KE, Walsh FS (1988) Distinct dystrophin mRNA species are expressed in embryonic and adult mouse skeletal muscle. FEBS Lett 242:47–52Google Scholar
  12. 12.
    Den Dunnen JT, Grootscholten PM Bakker E, Blonden LAJ, Ginjaar HB, Wapenaar MC, van Paassen HMB, van Broeckhoven C, Pearson PL, van Ommen GJB (1989) Topography of the Duchenne muscular dystrophy (DMD) gene: FIGE and cDNA analysis of 194 cases reveals 115 deletions and 13 duplications. Am J Hum Genet 45:835–847Google Scholar
  13. 13.
    Duchenne GB (1868) Recherches sur la paralysie musculaire pseudohypertrophique ou paralysie myo-schlérosique. Arch Gen Med 11: 5–25, 170–209, 305–321, 421–443, 552–588Google Scholar
  14. 14.
    Emery AEH (1988) Duchenne muscular dystrophy, revised edn. Oxford monographs on medical genetics no. 15. Oxford University PressGoogle Scholar
  15. 15.
    Emery AEH, Skinner R (1976) Clinical studies in benign (Becker type) X-linked muscular dystrophy. Clin Genet 10:189–201Google Scholar
  16. 16.
    England S, Nicholson LVB, Johnson MA, Forrest SM, Love D, Zubrzycka-Gaarn E, Bulman DE, Harris JB, Davies KE (1990) Very mild muscular dystrophy associated with the deletion of 46% of dystrophin. Nature 343:180–182Google Scholar
  17. 17.
    Feener CA, Koenig M, Kunkel LM (1989) Alternative splicing of human dystrophin mRNA, generates isoforms, at the carboxy terminus. Nature 338:509–511Google Scholar
  18. 18.
    Gillard EF, Chamberlain JS, Murphy EG, Duff CL, Smith B, Burghes AHM, Thompson MW, Sutherland J, Oss I, Bodrug SE, Klamut HJ, Ray PN, Worton RG (1989) Molecular and phenotypic analysis of patients with deletions within the deletion-rich region of the Duchenne muscular dystrophy (DMD) gene. Am J Hum Genet 45:507–520Google Scholar
  19. 19.
    Goodman SR, Krebs KE, Whitfield CF, Riederer BM, Zagon IS (1988) Spectrin and related molecules. CRC Crit Rev Biochem 23:171–234Google Scholar
  20. 20.
    Hodgson S, Hart K, Abbs S, Heckmatt J, Rodillo E, Bobrow M, Dubowitz V (1989) Correlation of clinical and deletion data in Duchenne and Becker muscular dystrophy. J Med Genet 26:682–693Google Scholar
  21. 21.
    Hoffman EP, Kunkel LM (1989) Dystrophin abnormalities in Duchenne/Becker muscular dystrophy. Neuron 2:1019–1029Google Scholar
  22. 22.
    Hoffman EP, Brown RH, Kunkel LM (1987) Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell 51:919–928Google Scholar
  23. 23.
    Hoffman EP, Fischbeck KH Brown RH Johnson M, Medori R, Loike JD, Harris JB, Waterston R, Brooke M, Specht L, Kupsky W, Chamberlein J, Caskey T, Shapiro F, Kunkel LM (1988) Characterization of Dystrophin in muscle-biopsy speciments from patients with Duchenne's or Becker's muscular dystrophy. N Engl J Med 318:1363–1368Google Scholar
  24. 24.
    Hoffman EP, Kunkel LM, Angelini C, Clarke A, Johnson M, Harris JB (1989) Improved diagnosis of Becker muscular dystrophy by dystrophin testing Neurology 39:1011–1017Google Scholar
  25. 25.
    Hoffman EP, Watkins SC, Slayter HS, Kunkel LM (1989) Detection of a specific isoform of alpha-actinin with antisera directed against dystrophin. J Cell Biol 108:503–510Google Scholar
  26. 26.
    Hori S, Sugiura H, Shimizu T, Hirabayashi T, Ohtani S, Yoshida M, Miyamoto K, Tanabe H (1989) Detection of dystrophin on two-dimentional gel electrophoresis. Biochem Biophys Res Commun 161:726–731Google Scholar
  27. 27.
    Kingston HM, Sarfarazi M, Thomas NST, Harper PS (1984) Localisation of the Becker muscular dystrophy gene on the short arm of the X chromosome by linkage to cloned DNA sequences. Hum Genet 67:6–17Google Scholar
  28. 28.
    Knudson CM, Hoffman EP, Kahl SD, Kunkel LM, Campbell KP (1988) Evidence for the association of dystrophin with the transverse tubular system in skeletal muscle. J Biol Chem 263:8480–8484Google Scholar
  29. 29.
    Koenig M, Hoffman EP, Bertelson CJ, Monaco AP, Feener C, Kunkel LM (1987) Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organisation of the DMD gene in normal and affected individuals. Cell 50:509–517Google Scholar
  30. 30.
    Koenig M, Monaco AP, Kunkel LM (1988) The complete sequence of dystrophin predicts a rod-shaped cryoskeletal protein. Cell 53:219–228Google Scholar
  31. 31.
    Koenig M plus 35 co-authors (1989) The molecular basis for Duchenne versus Becker muscular dystrophy: correlation of severity with type of deletion. Am J Hum Genet 45:498–506Google Scholar
  32. 32.
    Malhotra SB, Hart KA, Klamut HJ, Thomas NST, Bodrug SE, Burghes AHM, Bobrow M, Harper PS, Thompson MW, Ray PN, Worton RG (1988) Frame-shift deletions in patients with Duchenne and Becker muscular dystrophy. Science 242:755–759Google Scholar
  33. 33.
    Monaco AP Bertelson CJ, Liechti-Gallati S, Moser H, Kunkel LM (1988) An explanation for the phenotypic differences between patients bearing partial deletions of the DMD locus. Genomics 2:90–95Google Scholar
  34. 34.
    Muntoni F, Strong PN (1989) Transcription of the dystrophin gene in Duchenne muscular dystrophy muscle. FEBS Lett 252:95–98Google Scholar
  35. 35.
    Nicholson LVB, Davison K, Falkous G, Harwood C, O'Donnell E, Slater CR, Harris JB (1989) Dystrophin in skeletal muscle. I. Western blot analysis using a monoclonal antibody. J Neurol Sci 94:125–136Google Scholar
  36. 36.
    Nicholson LVB, Davison K, Johnson MA, Slater CR, Young C, Bhattacharya S Gardner-Medwin D, Harris JB (1989b) Dystrophin in skeletal muscle. II. Immunoreactivity, in patients with Xp21 muscular dystrophy. J Neurol Sci 94: 137–146Google Scholar
  37. 37.
    Nordenskjöld M Nicholson LVB, Edström L, Anvret M, Eiserman M, Slater CR, Stolpe L (1990) A normal male with an inherited deletion, of one exon within the DMD gene. Hum Genet 84:207–209Google Scholar
  38. 38.
    Nudel U, Zuk D, Einat P, Zeelon E, Levy Z, Neuman S, Yaffe D (1989) Duchenne muscular dystrophy gene product is not identical in muscle and brain. Nature 337:76–78Google Scholar
  39. 39.
    Patel K, Voit T, Dunn MJ, Strong PN, Dubowitz V (1988) Dystrophin and nebulin in the muscular dystrophies. J Neurol Sci 87:315–326Google Scholar
  40. 40.
    Ryder-Cook AS, Sicinski P, Thomas K, Davies KE, Worton RG Barnard EA, Darlison MG, Barnard PJ (1988) Localization of the mdx mutation within the mouse dystrophin gene. EMBO Journal 7:3017–3021Google Scholar
  41. 41.
    Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R Horn GT, Mullis KB, Erlich HA (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491Google Scholar
  42. 42.
    Shimizu T, Matsumura K, Hashimoto K, Mannen T Ishiguro T, Eguchi C, Nonaka I, Yoshida M, Ozawa E (1988) A monoclonal antibody against a synthetic polypeptide fragment of dystrophin (amino acid sequence from position 215 to 264). Proc Jpn Acad 64:205–208Google Scholar
  43. 43.
    Walton JN, Gardner-Medwin D (1988) The muscular dystrophies. In: Walton JN (ed) Disorders of voluntary muscle, 5th edn. Churchill Livingstone, Edinburgh pp 519–568Google Scholar
  44. 44.
    Watkins SC, Hoffman ED, Slayter HS, Kunkel LM (1988) Immunoelectron microscopic localization of dystrophin in myofibres. Nature 333:863–866Google Scholar
  45. 45.
    Worton RG, Thompson MW (1988) Genetics of Duchenne muscular dystrophy. Annu Rev Genet 22:601–629Google Scholar
  46. 46.
    Zubrzycka-Gaarn EE, Bulman DE, Karpati G, Burghes AHM, Belfall B, Klamut HJ, Talbot J, Hodges RS, Ray PN, Worton RG (1988) The Duchenne muscular dystrophy gene product is localized in sarcolemma of human skeletal muscle. Nature 333:466–469Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • L. V. B. Nicholson
    • 1
  • M. A. Johnson
    • 1
  • D. Gardner-Medwin
    • 1
  • S. Bhattacharya
    • 2
  • J. B. Harris
    • 1
  1. 1.Muscular Dystrophy Group Research Laboratories, Regional Neurological CentreNewcastle General HospitalNewcastle-upon-TyneGreat Britain
  2. 2.Department of Human GeneticsUniversity of Newcastle upon TyneNewcastle-upon-TyneGreat Britain

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