Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1981 Feb 1;88(2):329–337. doi: 10.1083/jcb.88.2.329

Structural changes in lysosomes from cultured human fibroblasts in Duchenne's muscular dystrophy

PMCID: PMC2111753  PMID: 6782112

Abstract

We have previously reported a decreased activity of the lysosomal enzyme dipeptidyl aminopeptidase-I (DAP-I) in cultured fibroblasts from patients with Duchenne's muscular dystrophy (DMD). Here we report that electron microscope examination of these cells reveals the presence of abundant lamellar bodies, a morphologic abnormalities commonly associated with impaired lysosomal function. Morphometric analysis of these cytoplasmic figures in dystrophic cells shows a sevenfold increase relative to normal controls (P less than 0.01). Analysis of lysosomal density profiles by density gradient centrifugation reveals similar patterns in normal and DMD cells. Treatment of lysosomes wit the nonionic detergent Triton X-100 causes an activation of DAP-I. This activation, attributable to structure-linked latency, is markedly diminished in DMD cells which show an optimal activation of only 180% compared to 255% for control fibroblasts (P less than 0.01). These data suggest an alteration in the properties of the lysosomal membrane in DMD fibroblasts. This suggestion is also supported by studies on the release of DAP-I from lysosomes by osmotic shock which show it to be a membrane-associated enzyme with membrane-binding characteristics intermediate between those of tightly bound beta-glucosidase and those of unbound N-acetylgalactosaminidase. The latency characteristics of these other lysosomal enzymes are not altered in the DMD cells, indicating that the effect is specific for DAP-I.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aubert-Tulkens G., Van Hoof F., Tulkens P. Gentamicin-induced lysosomal phospholipidosis in cultured rat fibroblasts. Quantitative ultrastructural and biochemical study. Lab Invest. 1979 Apr;40(4):481–491. [PubMed] [Google Scholar]
  2. Cullen M. J., Parsons R. Inclusion bodies in muscular dystrophy. Lancet. 1977 Oct 29;2(8044):929–929. doi: 10.1016/s0140-6736(77)90862-5. [DOI] [PubMed] [Google Scholar]
  3. Engel A. G. Evolution and content of vacuoles in primary hypokalemic periodic paralysis. Mayo Clin Proc. 1970 Nov-Dec;45(11):774–814. [PubMed] [Google Scholar]
  4. Gelman B. B., Papa L., Davis M. H., Gruenstein E. Decreased lysosomal dipeptidyl aminopeptidase I activity in cultured human skin fibroblasts in Duchenne's muscular dystrophy. J Clin Invest. 1980 Jun;65(6):1398–1406. doi: 10.1172/JCI109804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Goldman R. Ion distribution and membrane permeability in lysosomal suspensions. Front Biol. 1976;45:309–336. [PubMed] [Google Scholar]
  6. Hanai J., Leroy J., O'Brien J. S. Ultrastructure of cultured fibroblasts in I-cell disease. Am J Dis Child. 1971 Jul;122(1):34–38. doi: 10.1001/archpedi.1971.02110010070011. [DOI] [PubMed] [Google Scholar]
  7. Ionasescu V., Lara-Braud C., Zellweger H., Ionasescu R., Burmeister L. Fibroblast cultures in Duchenne muscular dystrophy. Alterations in synthesis and secretion of collagen and noncollagen proteins. Acta Neurol Scand. 1977 May;55(5):407–417. doi: 10.1111/j.1600-0404.1977.tb05659.x. [DOI] [PubMed] [Google Scholar]
  8. Kao I., Gordon A. M. Alteration of skeletal muscle cellular structures by potassium depletion. Neurology. 1977 Sep;27(9):855–860. doi: 10.1212/wnl.27.9.855. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Lie S. O., Schofield B. H., Taylor H. A., Jr, Doty S. B. Structure and function of the lysosomes of human fibroblasts in culture: dependence on medium pH. Pediatr Res. 1973 Jan;7(1):13–19. doi: 10.1203/00006450-197301000-00003. [DOI] [PubMed] [Google Scholar]
  11. Lipetz J., Cristofalo V. J. Ultrastructural changes accompanying the aging of human diploid cells in culture. J Ultrastruct Res. 1972 Apr;39(1):43–56. doi: 10.1016/s0022-5320(72)80005-4. [DOI] [PubMed] [Google Scholar]
  12. Lucky A. W., Mahoney M. J., Barrnett R. J., Rosenberg L. E. Electron microscopy of human skin fibroblasts in situ during growth in culture. Exp Cell Res. 1975 May;92(2):383–393. doi: 10.1016/0014-4827(75)90392-4. [DOI] [PubMed] [Google Scholar]
  13. Macdonald R. D., Engel A. G. Experimental chloroquine myopathy. J Neuropathol Exp Neurol. 1970 Jul;29(3):479–499. doi: 10.1097/00005072-197007000-00010. [DOI] [PubMed] [Google Scholar]
  14. Mawatari S., Miranda A., Rowland L. P. Adenyl cyclase abnormality in Duchenne muscular dystrophy: muscle cells in culture. Neurology. 1976 Nov;26(11):1021–1026. doi: 10.1212/wnl.26.11.1021. [DOI] [PubMed] [Google Scholar]
  15. Mawatari S., Schonberg M., Olarte M. Biochemical abnormalities of erythrocyte membranes in Duchenne dystrophy. Adenosine triphosphatase and adenyl cyclase. Arch Neurol. 1976 Jul;33(7):489–493. doi: 10.1001/archneur.1976.00500070031006. [DOI] [PubMed] [Google Scholar]
  16. Pena S. D., Vust A., Tucker D., Hamerton J. L., Wrogemann K. Biochemical investigations in cultured skin fibroblasts from patients with Duchenne muscular dystrophy. Clin Genet. 1978 Jul;14(1):50–54. doi: 10.1111/j.1399-0004.1978.tb02060.x. [DOI] [PubMed] [Google Scholar]
  17. Pickard N. A., Gruemer H. D., Verrill H. L., Isaacs E. R., Robinow M., Nance W. E., Myers E. C., Goldsmith B. Systemic membrane defect in the proximal muscular dystrophies. N Engl J Med. 1978 Oct 19;299(16):841–846. doi: 10.1056/NEJM197810192991601. [DOI] [PubMed] [Google Scholar]
  18. Reijngoud D. J., Tager J. M. The permeability properties of the lysosomal membrane. Biochim Biophys Acta. 1977 Nov 14;472(3-4):419–449. doi: 10.1016/0304-4157(77)90005-3. [DOI] [PubMed] [Google Scholar]
  19. Rome L. H., Garvin A. J., Allietta M. M., Neufeld E. F. Two species of lysosomal organelles in cultured human fibroblasts. Cell. 1979 May;17(1):143–153. doi: 10.1016/0092-8674(79)90302-7. [DOI] [PubMed] [Google Scholar]
  20. SAMUELS S., KOREY S. R., GONATAS J., TERRY R. D., WEISS M. Studies in Tay-Sachs disease. IV. Membranous cytoplasmic bodies. J Neuropathol Exp Neurol. 1963 Jan;22:81–97. [PubMed] [Google Scholar]
  21. SWIFT H., HRUBAN Z. FOCAL DEGRADATION AS A BIOLOGICAL PROCESS. Fed Proc. 1964 Sep-Oct;23:1026–1037. [PubMed] [Google Scholar]
  22. Stern J. The formation of sulfatide inclusions in organized nervous tissue culture. Lab Invest. 1973 Jan;28(1):87–95. [PubMed] [Google Scholar]
  23. Wyatt P. R., Cox D. M. Duchenne's muscular dystrophy: studies in cultured fibroblasts. Lancet. 1977 Jan 22;1(8004):172–174. doi: 10.1016/s0140-6736(77)91768-8. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES