Abstract
Our aim was to study how mouse skeletal muscle membranes are altered by eccentric and isometric contractions. A fluorescent dialkyl carbocyanine dye (DiOC18(3)) was used to label muscle membranes, and the membranes accessible to the dye were observed by confocal laser scanning microscopy. Experiments were done on normal mouse soleus muscles and soleus muscles injured by 20 eccentric or 20 isometric contractions. Longitudinal optical sections of control muscle fibers revealed DiOC18(3) staining of the plasmalemma and regularly spaced transverse bands corresponding in location to the T-tubular system. Transverse optical sections showed an extensive reticular network with the DiOC18(3) staining. Injured muscle fibers showed distinctively different staining patterns in both longitudinal and transverse optical sections. Longitudinal optical sections of the injured fibers revealed staining in a longitudinally-oriented pattern. No correlations were found between the abnormal DiOC18(3) staining and the reductions in maximal isometric tetanic force or release of lactate dehydrogenase (P≥0.32). Additionally, no difference in the extent of abnormal staining was found between muscles performing eccentric contractions and those performing the less damaging isometric contractions. However, many fibers in muscles injured by eccentric contractions showed swollen regions with marked loss of membrane integrity and an elevated free cytosolic calcium concentration as observed in Fluo-3 images. In conclusion, a loss of cell membrane integrity results from contractile activity, enabling DiOC18(3) staining of internal membranes. The resulting staining pattern is striking and fibers with damaged cell membranes are easily distinguished from uninjured ones.
Similar content being viewed by others
References
Armstrong RB, Warren GL, Warren JA (1991) Mechanisms of exercise-induced muscle fibre injury. Sports Med 12:184–207
Armstrong RB, Ogilvie RW, Schwane JA (1983) Eccentric exercise-induced injury to rat skeletal muscle. J Appl Physiol 54:80–93
Bartheid CS von, Cunningham DE, Rubel EW (1992) Neuronal tracing with DiI: decalcification, cryosetioning, and photoconversion for light and electron microscopic analysis. J Histochem Cytochem 38:725–733
Baumann O, Kitazawa T, Somylo AP (1990) Laser confocal scanning microscopy of the surface membrane/t-tubular system and the sarcoplasmic reticulum in insect striated muscle stained with DiIC18(3). J Struct Biol 105:154–161
Carpenter S, Karpati G (1989) Segmental necrosis and its demarcation in experimental micropuncture injury of skeletal muscle fibers. J Neuropathol Exp Neurol 48:154–170
Eisenberg BR (1983) Quantitative ultrastructure of mammalian skeletal muscle. In: Peachey LD, Adrian RH, Geiger SR (eds) Hanbook of Physiology. American Physiological Society, Bethesda, MD, pp 73–112
Flucher BE, Terasaki M, Chin H, Beeler TJ, Daniels MP (1991) Biogenesis of transverse tubules in skeletal muscle in vitro. Devl Biol 145:77–90
Godement P, Vanselow J, Thanos S, Bonhoeffer F (1987) A study in developing visual systems with a new method of staining neurones and their processes in fixed tissue. Development 101:697–713
Honig MC, Hume RI (1986) Fluorescent carbocyanine dyes allow living neurons of identified origin to be studied in long-term cultures. J Cell Biol 103:171–187
Lee RC, Gaylor DC, Bhatt D, Israel DA (1988) Role of cell membrane rupture in the pathogenesis of electrical trauma. J Surg Res 44:709–719
Liu DWC, Westerfield M (1992) Clustering of muscle acetylcholine receptors requires motoneurons in live embryos, but not in cell culture. J Neurosci 12:1859–1866
Lowe DA, Warren GL, Hayes DA, Farmer MA, Armstrong RB (1994) Eccentric contraction-induced injury of mouse soleus muscle: effect of varying [Ca2+]o. J Appl Physiol 76:1445–1453
McNeil PL, Khakee R (1992) Disruption of muscle fiber plasma membranes: role in exercise-induced damage. Am J Pathol 140:1097–1109
Peachey LD, Eisenberg BR (1978) Helicoids in the T system and striations of frog skeletal muscles seen by high voltage electron microscopy. Biophys J 22:145–154
Peachey LD, Franzini-Armstrong C (1983) Structure and function of membrane systems of skeletal muscle fibers. In: Peachey LD, Adrian RH, Geiger SR (eds) Handbook of Physiology, American Physiological Society, Bethesda, MD, pp 23–71
Ragnarson B, Bengtsson L, Hægerstrand A (1992) Labeling with fluorescent carbocyanine dyes of cultured endothelial and smooth muscle cells by growth in dye-containing medium. Histochemistry 97:329–333
Robertson TA, Papadimitriou JM, Grounds MD (1993) Fusion of myogenic cells to the newly sealed region of damaged myofibres in skeletal muscle regeneration. Neuropathol Appl Neurobiol 19:350–358
Simons K, Helenius A, Garoff H (1973) Solubilization of the membrane proteins from Semliki Forest virus with Triton X-100. J Molec Biol 80:119–133
Verrati E (1961) Investigations on the fine structure of striated muscle fiber. J Biophys Biochem Cytol 10:1–60
Warren GL, Hayes DA, Lowe DA, Armstrong RB (1993a) Mechanical factors in the initiation of eccentric contraction-induced injury in rat soleus muscle. J Physiol (Lond) 464:457–475
Warren GL, Hayes DA, Lowe DA, Prior BM, Armstrong RB (1993b) Materials fatigue initiates eccentric contraction-induced injury in rat soleus muscle. J Physiol (Lond) 464:477–489
Warren GL, Lowe DA, Hayes DA, Karwoski CJ, Prior BM, Armstrong RB (1993c) Excitation failure in eccentric contraction-induced injury of mouse soleus muscle. J Physiol (Lond) 468:487–499
Warren GL, Hayes DA, Lowe DA, Williams JH, Armstrong RB (1994) Eccentric contraction-induced injury in normal and hindlimb-suspended mouse soleus and EDL muscles. J Appl Physiol 77:1421–1430
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Warren, G.L., Lowe, D.A., Hayes, D.A. et al. Redistribution of cell membrane probes following contraction-induced injury of mouse soleus muscle. Cell Tissue Res 282, 311–320 (1995). https://doi.org/10.1007/BF00319121
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00319121