Letter | Published:

Defective membrane repair in dysferlin-deficient muscular dystrophy

Abstract

Muscular dystrophy includes a diverse group of inherited muscle diseases characterized by wasting and weakness of skeletal muscle1. Mutations in dysferlin are linked to two clinically distinct muscle diseases, limb-girdle muscular dystrophy type 2B and Miyoshi myopathy, but the mechanism that leads to muscle degeneration is unknown2,3. Dysferlin is a homologue of the Caenorhabditis elegans fer-1 gene, which mediates vesicle fusion to the plasma membrane in spermatids4. Here we show that dysferlin-null mice maintain a functional dystrophin–glycoprotein complex but nevertheless develop a progressive muscular dystrophy. In normal muscle, membrane patches enriched in dysferlin can be detected in response to sarcolemma injuries. In contrast, there are sub-sarcolemmal accumulations of vesicles in dysferlin-null muscle. Membrane repair assays with a two-photon laser-scanning microscope demonstrated that wild-type muscle fibres efficiently reseal their sarcolemma in the presence of Ca2+. Interestingly, dysferlin-deficient muscle fibres are defective in Ca2+-dependent sarcolemma resealing. Membrane repair is therefore an active process in skeletal muscle fibres, and dysferlin has an essential role in this process. Our findings show that disruption of the muscle membrane repair machinery is responsible for dysferlin-deficient muscle degeneration, and highlight the importance of this basic cellular mechanism of membrane resealing in human disease.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Cohn, R. D. & Campbell, K. P. Molecular basis of muscular dystrophies. Muscle Nerve 23, 1456–1471 (2000)

  2. 2

    Bashir, R. et al. A gene related to Caenorhabditis elegans spermatogenesis factor fer-1 is mutated in limb-girdle muscular dystrophy type 2B. Nature Genet. 20, 37–42 (1998)

  3. 3

    Liu, J. et al. Dysferlin, a novel skeletal muscle gene, is mutated in Miyoshi myopathy and limb girdle muscular dystrophy. Nature Genet. 20, 31–36 (1998)

  4. 4

    Achanzar, W. E. & Ward, S. A nematode gene required for sperm vesicle fusion. J. Cell Sci. 110, 1073–1081 (1997)

  5. 5

    Piccolo, F., Moore, S. A., Ford, G. C. & Campbell, K. P. Intracellular accumulation and reduced sarcolemmal expression of dysferlin in limb-girdle muscular dystrophies. Ann. Neurol. 48, 902–912 (2000)

  6. 6

    Matsuda, C. et al. The sarcolemmal proteins dysferlin and caveolin-3 interact in skeletal muscle. Hum. Mol. Genet. 10, 1761–1766 (2001)

  7. 7

    Anderson, L. V. et al. Secondary reduction in calpain 3 expression in patients with limb girdle muscular dystrophy type 2B and Miyoshi myopathy (primary dysferlinopathies). Neuromusc. Disord. 10, 553–559 (2000)

  8. 8

    Straub, V., Rafael, J. A., Chamberlain, J. S. & Campbell, K. P. Animal models for muscular dystrophy show different patterns of sarcolemmal disruption. J. Cell Biol. 139, 375–385 (1997)

  9. 9

    Duclos, F. et al. Progressive muscular dystrophy in α-sarcoglycan-deficient mice. J. Cell Biol. 142, 1461–1471 (1998)

  10. 10

    Durbeej, M. & Campbell, K. P. Muscular dystrophies involving the dystrophin–glycoprotein complex: an overview of current mouse models. Curr. Opin. Genet. Dev. 12, 349–361 (2002)

  11. 11

    Ervasti, J. M., Ohlendieck, K., Kahl, S. D., Gaver, M. G. & Campbell, K. P. Deficiency of a glycoprotein component of the dystrophin complex in dystrophic muscle. Nature 345, 315–319 (1990)

  12. 12

    Petrof, B. J., Shrager, J. B., Stedman, H. H., Kelly, A. M. & Sweeney, H. L. Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc. Natl Acad. Sci. USA 90, 3710–3714 (1993)

  13. 13

    Clarke, M. S., Khakee, R. & McNeil, P. L. Loss of cytoplasmic basic fibroblast growth factor from physiologically wounded myofibers of normal and dystrophic muscle. J. Cell Sci. 106, 121–133 (1993)

  14. 14

    Coral-Vazquez, R. et al. Disruption of the sarcoglycan–sarcospan complex in vascular smooth muscle: a novel mechanism for cardiomyopathy and muscular dystrophy. Cell 98, 465–474 (1999)

  15. 15

    McNeil, P. L. & Khakee, R. Disruptions of muscle fiber plasma membranes. Role in exercise-induced damage. Am. J. Pathol. 140, 1097–1109 (1992)

  16. 16

    McNeil, P. L. & Terasaki, M. Coping with the inevitable: how cells repair a torn surface membrane. Nature Cell Biol. 3, E124–E129 (2001)

  17. 17

    Bi, G. Q., Alderton, J. M. & Steinhardt, R. A. Calcium-regulated exocytosis is required for cell membrane resealing. J. Cell Biol. 131, 1747–1758 (1995)

  18. 18

    Miyake, K. & McNeil, P. L. Vesicle accumulation and exocytosis at sites of plasma membrane disruption. J. Cell Biol. 131, 1737–1745 (1995)

  19. 19

    Steinhardt, R. A., Bi, G. & Alderton, J. M. Cell membrane resealing by a vesicular mechanism similar to neurotransmitter release. Science 263, 390–393 (1994)

  20. 20

    Reddy, A., Caler, E. V. & Andrews, N. W. Plasma membrane repair is mediated by Ca2+-regulated exocytosis of lysosomes. Cell 106, 157–169 (2001)

  21. 21

    Davis, D. B., Doherty, K. R., Delmonte, A. J. & McNally, E. M. Calcium-sensitive phospholipid binding properties of normal and mutant ferlin C2 domains. J. Biol. Chem. 277, 22883–22888 (2002)

  22. 22

    McNeil, P. L., Vogel, S. S., Miyake, K. & Terasaki, M. Patching plasma membrane disruptions with cytoplasmic membrane. J. Cell Sci. 113, 1891–1902 (2000)

  23. 23

    McNeil, P. L., Miyake, K. & Vogel, S. S. The self-sealing membrane concept. Proc. Natl Acad. Sci. USA 100, 4592–4597 (2003)

  24. 24

    Ohlendieck, K. & Campbell, K. P. Dystrophin-associated proteins are greatly reduced in skeletal muscle from mdx mice. J. Cell Biol. 115, 1685–1694 (1991)

  25. 25

    Durbeej, M. et al. Disruption of the β-sarcoglycan gene reveals pathogenetic complexity of limb-girdle muscular dystrophy type 2E. Mol. Cell 5, 141–151 (2000)

  26. 26

    Anderson, L. V. et al. Dysferlin is a plasma membrane protein and is expressed early in human development. Hum. Mol. Genet. 8, 855–861 (1999)

  27. 27

    Miner, J. H. et al. The laminin alpha chains: expression, developmental transitions, and chromosomal locations of α1-5, identification of heterotrimeric laminins 8–11, and cloning of a novel α3 isoform. J. Cell Biol. 137, 685–701 (1997)

Download references

Acknowledgements

We thank all members of the Campbell laboratory for the critical reading of this manuscript, discussions and supplying critical reagents; and M. Hassebrock, S. Lowen, E. Hurst and K. Garringer for technical assistance. D. Bansal was supported by American Heart Predoctoral Fellowship (Heartland). We thank the University of Iowa DNA Core Facility, which is supported in part by the Diabetes Endocrinology Research Center and the University of Iowa Roy J. and Lucille A. Carver College of Medicine. We also thank University of Iowa Central Microscopy Research Facility and the Medical College of Georgia Imaging Core. This work was supported by the Muscular Dystrophy Association (K.P.C.). P.L.McN. is supported by NASA, and S.S.V. by NIH. K.P.C. is an investigator of the Howard Hughes Medical Institute.

Author information

Correspondence to Kevin P. Campbell.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Movie 1 (AVI 151 kb)

Supplementary Movie 2 (AVI 156 kb)

Supplementary Figure 1 (JPG 73 kb)

Supplementary Figure 2 (JPG 430 kb)

Supplementary Figure 3 (JPG 249 kb)

Supplementary Movie 3 (AVI 153 kb)

Supplementary Movie 4 (AVI 184 kb)

Supplementary Legends (DOC 27 kb)

Erratum to supplementary information (DOC 24 kb)

Rights and permissions

Reprints and Permissions

About this article

Further reading

Figure 1: Complete loss of dysferlin expression in dysferlin-null mice.
Figure 2: Dysferlin-null mice develop progressive muscular dystrophy.
Figure 3: Normal expression of DGC components and a structurally stable sarcolemma in dysferlin-null mice.
Figure 4: Vesicle accumulation and dysferlin-enriched membrane patches on the damaged muscle fibres.
Figure 5: Defective resealing of membranes in dysferlin-null muscle.

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.