User menu

Accès à distance ? S'identifier sur le proxy UCLouvain

Long-term study of Ca(2+) homeostasis and of survival in collagenase-isolated muscle fibres from normal and mdx mice.

Primary tabs

De Backer, Fabienne [UCL] Vandebrouck, Clarisse [UCL] Gailly, Philippe [UCL] Gillis, Jean-Marie [UCL]

Skeletal muscles of the mdx mouse lack dystrophin offering the possibility to study the role of intracellular Ca(2+) ions in fibre degeneration. Flexor digitorum brevis muscles of 3-month-old mdx and normal mice were dissociated with collagenase; fibres were maintained in culture for 6 days (d0 to d5) and their survival was assessed. Cytosolic [Ca(2+)], passive Mn(2+) influx (indicative of Ca(2+) influx) and activity of mechanosensitive/voltage-independent Ca(2+) channels were studied over the same period. Survival of normal fibres declined steadily from d0 to d3, but an acceleration of fibre death occurred in mdx fibres from d1 to d2. This could be greatly reduced but not abolished by lowering external [Ca(2+)] 10-fold. In the d0-d5 period, both mdx and normal fibres showed transient increases of Mn(2+) influx and activity of the Ca(2+) channels; these peaked at d1 and disappeared by d3-d4. Increases were always significantly larger in mdx fibres. Altogether, over the 6 days, 130 paired measurements of [Ca(2+)](i) and Mn(2+) influx were made on 68 fibres from mdx and 62 fibres from normal mice. In 90 % of the fibres, [Ca(2+)](i) remained within the 25-85 nM limits while Mn(2+) influx varied more than 10-fold. The median for Mn(2+) influx was 45 % greater in fibres from mdx mice than in fibres from control C57 mice. However, there was no significant difference between [Ca(2+)](i) medians in fibres from normal and mdx mice. Addition of 25-75 nM of a Ca(2+) ionophore (4-bromo-A23187) to the medium did not affect the level of cytosolic [Ca(2+)] in both types of fibres, while markedly increasing the rate of Mn(2+) influx, as expected. Thus, Ca(2+) homeostasis was equally robust in mdx and normal fibres. The remaining 10 % of the fibres showed, at d1, high levels of Mn(2+) influx and/or elevated [Ca(2+)](i) above 100 nM. This did not affect survival of normal fibres but was probably responsible of the increased death rate in mdx fibres.

Document type Article de périodique (Journal article) – Journal Article, Research Support, Non-U.S. Gov't
Access type Accès restreint
Publication date 2002
Journal information "The Journal of physiology" - Vol. 542, no. Pt 3, p. 855-65 (2002)
Peer reviewed yes
issn 0022-3751
Publication status Publié
Affiliations UCL - MD/FSIO - Département de physiologie et pharmacologie
UCL - MD/NOPS - Département de neurologie et de psychiatrie
MESH Subject Animals ; Calcium - metabolism ; Calcium Channels - physiology ; Collagenases ; Cytosol - metabolism ; Electrophysiology ; Histological Techniques ; Homeostasis ; Manganese - pharmacokinetics ; Mice ; Mice, Inbred C57BL ; Mice, Inbred mdx ; Muscle Fibers - metabolism - physiology ; Muscle, Skeletal - metabolism - physiopathology ; Muscular Dystrophy, Animal - metabolism - physiopathology ; Osmolar Concentration ; Reference Values ; Time Factors ; Tissue Survival
Links
  1. Bockhold Kathryn J., David Rosenblatt J., Partridge Terence A., Aging normal and dystrophic mouse muscle: Analysis of myogenicity in cultures of living single fibers, 10.1002/(sici)1097-4598(199802)21:2<173::aid-mus4>3.0.co;2-8
  2. Boland B., Himpens B., Denef J. F., Gillis J. M., Site-dependent pathological differences in smooth muscles and skeletal muscles of the adult MDX mouse, 10.1002/mus.880180612
  3. Collet Claude, Allard Bruno, Tourneur Yves, Jacquemond Vincent, Intracellular calcium signals measured with indo-1 in isolated skeletal muscle fibres from control andmdxmice, 10.1111/j.1469-7793.1999.00417.x
  4. Franco-Obregón A, Lansman J B, Mechanosensitive ion channels in skeletal muscle from normal and dystrophic mice., 10.1113/jphysiol.1994.sp020440
  5. Gailly Ph., Boland B., Himpens B., Casteels R., Gillis J.M., Critical evaluation of cytosolic calcium determination in resting muscle fibres from normal and dystrophic (mdx) mice, 10.1016/0143-4160(93)90006-r
  6. Gillis J. M., 10.1023/a:1005545325254
  7. GrynkiewicZ, Journal of Biological Chemistry, 260, 3440 (1985)
  8. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J., Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches, 10.1007/bf00656997
  9. Head S I, Membrane potential, resting calcium and calcium transients in isolated muscle fibres from normal and dystrophic mice., 10.1113/jphysiol.1993.sp019801
  10. Hopf, American Journal of Physiology, 271, C1325 (1996)
  11. Lee K S, Tsien R W, High selectivity of calcium channels in single dialysed heart cells of the guinea-pig., 10.1113/jphysiol.1984.sp015374
  12. Liu, American Journal of Physiology, 272, C1919 (1997)
  13. McCarter G.C., Steinhardt R.A., Increased Activity of Calcium Leak Channels Caused by Proteolysis Near Sarcolemmal Ruptures, 10.1007/s002320001086
  14. Mallouk N., Jacquemond V., Allard B., Elevated subsarcolemmal Ca2+ in mdx mouse skeletal muscle fibers detected with Ca2+-activated K+ channels, 10.1073/pnas.97.9.4950
  15. Merritt, Journal of Biological Chemistry, 264, 1522 (1989)
  16. Midrio M., Danieli-Betto Daniela, Megighian Aram, Betto Romeo, Early effects of denervation on sarcoplasmic reticulum properties of slow-twitch rat muscle fibres, 10.1007/s004240050413
  17. Moens P., Baatsen P. H. W. W., Mar�chal G., Increased susceptibility of EDL muscles from mdx mice to damage induced by contractions with stretch, 10.1007/bf00121296
  18. Pastoret Christian, Sebille Alain, mdx mice show progressive weakness and muscle deterioration with age, 10.1016/0022-510x(94)00276-t
  19. 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., 10.1073/pnas.90.8.3710
  20. Pressmar J., Brinkmeier H., Seewald M. J., Naumann T., R�del R., Intracellular Ca2+ concentrations are not elevated in resting cultured muscle from Duchenne (DMD) patients and in MDX mouse muscle fibres, 10.1007/bf00378527
  21. Sicinski P, Geng Y, Ryder-Cook A., Barnard E., Darlison M., Barnard P., The molecular basis of muscular dystrophy in the mdx mouse: a point mutation, 10.1126/science.2662404
  22. Turner P. R., Increased calcium influx in dystrophic muscle, 10.1083/jcb.115.6.1701
  23. Turner PaulR., Schultz Randon, Ganguly Bishu, Steinhardt RichardA., Proteolysis results in altered leak channel kinetics and elevated free calcium in mdx muscle, 10.1007/bf00232023
  24. Turner Paul R., Westwood Tim, Regen Chester M., Steinhardt Richard A., Increased protein degradation results from elevated free calcium levels found in muscle from mdx mice, 10.1038/335735a0
  25. Tutdibi O., Brinkmeier H., Rüdel R., Föhr K. J., Increased calcium entry into dystrophin-deficient muscle fibres of MDX and ADR-MDX mice is reduced by ion channel blockers, 10.1111/j.1469-7793.1999.859ab.x
Bibliographic reference De Backer, Fabienne ; Vandebrouck, Clarisse ; Gailly, Philippe ; Gillis, Jean-Marie. Long-term study of Ca(2+) homeostasis and of survival in collagenase-isolated muscle fibres from normal and mdx mice.. In: The Journal of physiology, Vol. 542, no. Pt 3, p. 855-65 (2002)
Permanent URL http://hdl.handle.net/2078.1/9213