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PMID: 7865881 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Myotubes from transgenic mdx mice expressing full-length dystrophin show normal calcium regulation.

Molecular biology of the cell ·Vol. 5 ·No. 10 ·1994-10-00 ·Pages 1159-67

Denetclaw WF, Hopf FW, Cox GA, Chamberlain JS, Steinhardt RA

Abstract

A lack of dystrophin results in muscle degeneration in Duchenne muscular dystrophy. Dystrophin-deficient human and mouse muscle cells have higher resting levels of intracellular free calcium ([Ca2+]i) and show a related increase in single-channel open probabilities of calcium leak channels. Elevated [Ca2+]i results in high levels of calcium-dependent proteolysis, which in turn increases calcium leak channel activity. This process could initiate muscle degeneration by further increasing [Ca2+]i and proteolysis in a positive feedback loop. Here, we tested the direct effect of restoration of dystrophin on [Ca2+]i and channel activity in primary myotubes from mdx mice made transgenic for full-length dystrophin. Transgenic mdx mice have been previously shown to have normal dystrophin localization and no muscle degeneration. Fura-2 calcium measurements and single-channel patch recordings showed that resting [Ca2+]i levels and open probabilities of calcium leak channels of transgenic mdx myotubes were similar to normal levels and significantly lower than mdx littermate controls (mdx) that lack dystrophin. Thus, restoration of normal calcium regulation in transgenic mdx mice may underlie the resulting absence of degeneration.

MeSH Terms
Animals Calcium/metabolism Calcium Channels/metabolism Cells, Cultured Dystrophin/genetics,metabolism Female Genetic Therapy Humans Male Mice Mice, Inbred C57BL Mice, Inbred mdx Mice, Transgenic Muscle, Skeletal/metabolism Muscular Dystrophy, Animal/genetics,metabolism,therapy
Chemicals
Calcium Channels Dystrophin Calcium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Denetclaw W F
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Hopf F W
Cox G A
Chamberlain J S
Steinhardt R A
References (46)
46 references, click to expand
  1. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.
    Pflugers Arch. 1981 Aug;391(2):85-100 PMID: 6270629
  2. Dystrophin abnormalities in Duchenne/Becker muscular dystrophy.
    Neuron. 1989 Jan;2(1):1019-29 PMID: 2696500
  3. Deficiency of a glycoprotein component of the dystrophin complex in dystrophic muscle.
    Nature. 1990 May 24;345(6273):315-9 PMID: 2188135
  4. Increased activity of calcium leak channels in myotubes of Duchenne human and mdx mouse origin.
    Science. 1990 Nov 2;250(4981):673-6 PMID: 2173137
  5. Intracellular ion activities and membrane transport in parietal cells measured with fluorescent dyes.
    Methods Enzymol. 1990;192:38-81 PMID: 2074799
  6. Effects of calcium on protein turnover of incubated muscles from mdx mice.
    Am J Physiol. 1991 Apr;260(4 Pt 1):E594-8 PMID: 2018123
  7. Non-ligand-activated chloride channels of skeletal muscle and epithelia.
    Prog Biophys Mol Biol. 1989;54(1):59-79 PMID: 2562293
  8. Total ion content of skeletal and cardiac muscle in the mdx mouse dystrophy: Ca2+ is elevated at all ages.
    J Neurol Sci. 1991 Jun;103(2):226-31 PMID: 1880541
  9. Fiber regeneration is not persistent in dystrophic (MDX) mouse skeletal muscle.
    Dev Biol. 1991 Nov;148(1):314-21 PMID: 1936568
  10. Discrimination of Ca(2+)-ATPase activity of the sarcoplasmic reticulum from actomyosin-type ATPase activity of myofibrils in skinned mammalian skeletal muscle fibres: distinct effects of cyclopiazonic acid on the two ATPase activities.
    J Muscle Res Cell Motil. 1991 Aug;12(4):355-65 PMID: 1834695
  11. Dystrophin-associated proteins are greatly reduced in skeletal muscle from mdx mice.
    J Cell Biol. 1991 Dec;115(6):1685-94 PMID: 1757468
  12. Increased calcium influx in dystrophic muscle.
    J Cell Biol. 1991 Dec;115(6):1701-12 PMID: 1661733
  13. Primary structure of dystrophin-associated glycoproteins linking dystrophin to the extracellular matrix.
    Nature. 1992 Feb 20;355(6362):696-702 PMID: 1741056
  14. Is the carboxyl-terminus of dystrophin required for membrane association? A novel, severe case of Duchenne muscular dystrophy.
    Ann Neurol. 1991 Oct;30(4):605-10 PMID: 1789686
  15. Regulation of intracellular free calcium in normal and dystrophic mouse cerebellar neurons.
    Brain Res. 1992 Apr 24;578(1-2):49-54 PMID: 1511288
  16. Single-channel recordings of chloride currents in cultured human skeletal muscle.
    Pflugers Arch. 1992 Jun;421(2-3):108-16 PMID: 1382261
  17. Diversity of airway epithelial cell targets for in vivo recombinant adenovirus-mediated gene transfer.
    J Clin Invest. 1993 Jan;91(1):225-34 PMID: 8423221
  18. Restoration of dystrophin-associated proteins in skeletal muscle of mdx mice transgenic for dystrophin gene.
    FEBS Lett. 1993 Apr 12;320(3):276-80 PMID: 8462697
  19. Ca2+ levels in myotubes grown from the skeletal muscle of dystrophic (mdx) and normal mice.
    J Physiol. 1993 Jan;460:1-13 PMID: 8487190
  20. Is the maintainance of the C-terminus domain of dystrophin enough to ensure a milder Becker muscular dystrophy phenotype?
    Hum Mol Genet. 1993 Jan;2(1):39-42 PMID: 8490621
  21. Myoblast transfer in Duchenne muscular dystrophy.
    Ann Neurol. 1993 Jul;34(1):8-17 PMID: 8517684
  22. Proteolysis results in altered leak channel kinetics and elevated free calcium in mdx muscle.
    J Membr Biol. 1993 May;133(3):243-51 PMID: 8392585
  23. A role for the dystrophin-glycoprotein complex as a transmembrane linker between laminin and actin.
    J Cell Biol. 1993 Aug;122(4):809-23 PMID: 8349731
  24. Overexpression of dystrophin in transgenic mdx mice eliminates dystrophic symptoms without toxicity.
    Nature. 1993 Aug 19;364(6439):725-9 PMID: 8355788
  25. Critical evaluation of cytosolic calcium determination in resting muscle fibres from normal and dystrophic (mdx) mice.
    Cell Calcium. 1993 Jun;14(6):473-83 PMID: 8358771
  26. Heterokaryon myotubes with normal mouse and Duchenne nuclei exhibit sarcolemmal dystrophin staining and efficient intracellular free calcium control.
    Mol Biol Cell. 1993 Sep;4(9):963-72 PMID: 8257798
  27. Membrane potential, resting calcium and calcium transients in isolated muscle fibres from normal and dystrophic mice.
    J Physiol. 1993 Sep;469:11-9 PMID: 8271194
  28. Intracellular Ca2+ concentrations are not elevated in resting cultured muscle from Duchenne (DMD) patients and in MDX mouse muscle fibres.
    Pflugers Arch. 1994 Apr;426(6):499-505 PMID: 8052519
  29. Muscle calcium and magnesium content in Duchenne muscular dystrophy.
    Neurology. 1982 Oct;32(10):1088-92 PMID: 6889695
  30. X chromosome-linked muscular dystrophy (mdx) in the mouse.
    Proc Natl Acad Sci U S A. 1984 Feb;81(4):1189-92 PMID: 6583703
  31. A new generation of Ca2+ indicators with greatly improved fluorescence properties.
    J Biol Chem. 1985 Mar 25;260(6):3440-50 PMID: 3838314
  32. Measurement of cytosolic free Ca2+ in individual small cells using fluorescence microscopy with dual excitation wavelengths.
    Cell Calcium. 1985 Apr;6(1-2):145-57 PMID: 3874696
  33. The Ca signal from fura-2 loaded mast cells depends strongly on the method of dye-loading.
    FEBS Lett. 1985 Nov 11;192(1):13-8 PMID: 3840439
  34. Calcium rises abruptly and briefly throughout the cell at the onset of anaphase.
    Science. 1986 Aug 22;233(4766):886-9 PMID: 3755550
  35. Muscular dystrophy in the mdx mouse: histopathology of the soleus and extensor digitorum longus muscles.
    J Neurol Sci. 1987 Aug;80(1):39-54 PMID: 3612180
  36. Fura-2 measurement of cytosolic free Ca2+ in monolayers and suspensions of various types of animal cells.
    J Cell Biol. 1987 Nov;105(5):2145-55 PMID: 3680375
  37. Dystrophin: the protein product of the Duchenne muscular dystrophy locus.
    Cell. 1987 Dec 24;51(6):919-28 PMID: 3319190
  38. Free cytoplasmic Ca++ at rest and after cholinergic stimulus is increased in cultured muscle cells from Duchenne muscular dystrophy patients.
    Neurology. 1988 Mar;38(3):476-80 PMID: 3347352
  39. Increased protein degradation results from elevated free calcium levels found in muscle from mdx mice.
    Nature. 1988 Oct 20;335(6192):735-8 PMID: 3173492
  40. Myoplasmic (Ca2+) in Duchenne muscular dystrophy patients.
    Acta Cient Venez. 1987;38(4):503-4 PMID: 3506369
  41. Association of dystrophin and an integral membrane glycoprotein.
    Nature. 1989 Mar 16;338(6212):259-62 PMID: 2493582
  42. Satellite cells from dystrophic (mdx) mouse muscle are stimulated by fibroblast growth factor in vitro.
    Differentiation. 1988 Nov;39(1):42-9 PMID: 3246292
  43. The molecular basis of muscular dystrophy in the mdx mouse: a point mutation.
    Science. 1989 Jun 30;244(4912):1578-80 PMID: 2662404
  44. Modification of L-type calcium current by intracellularly applied trypsin in guinea-pig ventricular myocytes.
    J Physiol. 1988 Oct;404:259-74 PMID: 2855349
  45. Regulation of cytosolic Ca2+ in clonal human muscle cell cultures.
    Biochem Biophys Res Commun. 1989 Dec 29;165(3):1130-7 PMID: 2532882
  46. Calcium entry through stretch-inactivated ion channels in mdx myotubes.
    Nature. 1990 Apr 12;344(6267):670-3 PMID: 1691450
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
Published
1994-10-00
Pages
1159-67
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC301138
Subset
IM
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