To study the physiological aspects of the excitation-contraction cycle, saponin (10–100Μg ml−1) was used as a skinning agent on muscle and sarcotubular vesicles derived from fast muscles (sartorius and tibialis anterior) ofRana esculenta.
The vesicles showed similar Ca2+-ATPase activity and similar protein profiles carried out by SDS-PAGE. Calcium transport in untreated vesicles and those treated with different concentrations of saponin seemed to have the same quantitative and qualitative parameters if the saponin was used in a range between 10 and 50Μg ml−1.
Our results confirm that saponin may be considered to be a valid skinning agent for the external membranes of fast skeletal muscles.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Altschuld, R., Hohl, C., Ansel, A. &Brierley, G. P. (1981) Compartimentation of K+ in isolated adult rat heart cells.Arch. Biochem. Biophys 209, 175–84.
Bolitho-Donaldson, S. K. (1985) Peeled mammalian skeletal muscle fibres: possible stimulation of Ca++ release via a transverse tubule sarcoplasmic reticulum mechanism.J. gen. Physiol. 86, 501–26.
Briggs, F. N., Poland, J. L. &Solaro, R. J. (1977) Relative capabilities of Sarcoplasmic reticulum in fast and slow mammalian skeletal muscle.J. Physiol. 266, 587–94.
Endo, M. (1977) Calcium release from the sarcoplasmic reticulum.Physiol. Rev. 57, 71–108.
Endo, M. &Kitazawa, T. (1978) E-C coupling studies on skinned cardiac fibres. InBiophysical Aspects of Cardiac Muscle (edited byMorad, M.), pp. 307–327. New York: Academic Press.
Endo, M. &Iino, M. (1980) Specific perforation of muscle cell membranes with preserved SR functions by saponin treatment.J. Musc. Res. Cell Mot. 1, 89–100.
Fitts, R. H., Winder, W. W., Brooke, M. H., Kaiser, K. K. &Holloszy, J. O. (1980) Contractile, biochemical and histochemical properties of thyrotoxic rat soleus muscle.Am. J. Physiol. 238, C15-C20.
Frank, G. B. (1982) Role of extracellular and ‘trigger“ calcium ions in excitation-contraction coupling in skeletal muscle.Can. J. Physiol. Phararnacol. 60, 427–39.
Kargacin, G. J. &Fay, S. F. (1985) Contractile properties of saponin-treated single smooth muscle cells.J. gen. Physiol. 86, 38a (Abs).
Lowry, O. H., Rosebrough, N. J., Farr, A. L. &Randall, R. J. (1951) Protein measurement with the Folin phenol reagent.J. biol. Chem. 193, 265–7.
Mancinelli, L., Fano', G., Ferroni, L., Secca, T. &Dolcini, B. M. (1983) Evidence for an inotropic positive action of cGMP during excitation-contraction coupling in frog sartorius muscle.Can. J. Physiol. Pharmacol. 61, 590–4.
Martonosi, A. (1968) Sarcoplasmic reticulum V: The structure of sarcoplasmic reticulum membranes.Biochem. Biophys. Acta 150, 694–704.
Miller, D. J. &Smith, G. L. (1985) The contractile behaviour of EGTA and detergent-treated heart muscle.J. Musc. Res. Cell Mot. 6, 541–67.
Ohtsuki, I., Manzi, R. M., Palade, G. E. &Jamieson, J. D. (1978) Entry of macromolecular tracers into cells fixed with low concentration of aldehydes.Biol. Cellulaire 31, 119–26.
Saini, M. S. &Van Etten, R. L. (1978) A homogeneous isoenzyme of human liver acid phosphates.Arch. Biochem. Biophys. 191, 613–24.
Saito, A., Seiler, S., Chu, A. &Fleischer, S. (1984) Preparation and morphology of sarcoplasmic reticulum terminal cisternae from rabbit skeletal muscle.J. cell. Biol. 99, 875–85.
Stephenson, E. W. (1981) Activation of fast skeletal muscle: contribution of studies on skinned fibres.Am. J. Physiol. 240, C1-C19.
Weber, K. &Osborne, M. (1969) For the reliability of molecular weight determinations by dodecyl sulphate polyacrylamide gel electrophoresis.J. biol. Chem. 224, 4406–12.
About this article
Cite this article
Fanó, G., Belia, S., Fulle, S. et al. Functional aspects of calcium transport in sarcoplasmic reticulum vesicles derived from frog skeletal muscle treated with saponin. J Muscle Res Cell Motil 10, 326–330 (1989). https://doi.org/10.1007/BF01758428
- Skeletal Muscle
- ATPase Activity
- Sarcoplasmic Reticulum