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In vitro studies of skeletal muscle membranes

Effects of denervation on the macromolecular components of cation transport in red and white skeletal muscle

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Summary

The effects of denervation on the macromolecular components of active monovalent cation transport in skeletal muscle have been studied using purified sarcolemma membranes. A comparison of membrane activities of fast-twitch, slow-twitch, and mixed-fiber muscles was made to determine what role, if any, the motor nerve has in regulating this important aspect of muscle metabolism. A dramatic increase in the basal sarcolemmal Mg++ ATPase activity (three- to fourfold) was found for both major muscle types. An increase in the ouabain-inhibitable (Na++K+)-stimulated enzyme was also found, but the effect was substantially less (1.5- to twofold). [3H]-ouabain binding, as an index of glycoside receptor sites, also increased (two- to threefold) midway in the course of denervation. On the other hand, the phosphorylated intermediate activity, a functional component of the transport system, clearly decreased over the same time course and remained below control values for the remainder of the course. This resulted in a two- to threefold increase in the turnover number, suggesting that active transport of cations should increase dramatically with denervation. The membrane protein patterns on SDS gels were less obvious than the changes observed in the functional components. The major effects appeared after only one week and seemed to be restricted to high molecular weight membrane proteins, especially in the 100,000 to 250,000 daltons range. This effect was more prominent in slow-twitch membranes with an apparent semiquantitative decrease in stain at 240,000 daltons. In gels of membranes from fast-twitch muscles a decreased stain in the range of 100,000 to 110,000 daltons occurred, and this became more obvious with longer periods of denervation. The results suggest that considerable influence on the macromolecular components of active cation transport in skeletal muscle is exerted by the motor nerve. No appreciable difference was found in this effect when the two major types of skeletal muscle, fast-twitch and slow-twitch, were compared, suggesting that motor nerve regulation of this membrane property is qualitatively the same.

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References

  • Albers, R. W. 1976. The (sodium plus potassium)-transport ATPase.In: The Enzymes of Biological Membranes. A. Martonosi, editor. Vol. 3, p. 283. Plenum Press, New York

    Google Scholar 

  • Albers, R.W., Koval, G.J., Siegel, G.J. 1968. Studies on the interaction of ouabain and other cardioactive steroids with sodium-potassium-activated adenosine triphosphatase.Mol. Pharmacol. 4:324

    PubMed  Google Scholar 

  • Albuquerque, E.X., Schuh, R.T., Kaufman, F.C. 1971. Early membrane depolarization of the fast mammalian muscle after denervation.Pfluegers Arch. Gesamte Physiol. 328:36

    Article  Google Scholar 

  • Andrew, C.G., Almon, R.R., Appel, S.H. 1974. Macromolecular characterization of muscle membranes. Acetylcholine receptor of normal and denervated muscle.J. Biol. Chem. 249:6163

    PubMed  Google Scholar 

  • Andrew, C.G., Almon, R.R., Appel, S.H. 1975. Macro-molecular characterization of muscle membranes. Endogenous membrane kinase and phosphorylated protein substrate from normal and denervated muscle.J. Biol. Chem. 250:3972

    PubMed  Google Scholar 

  • Andrew, C.G., Appel, S.H. 1973, Macromolecular characterization of muscle membranes. I. Proteins and sialic acid of normal and denervated muscle.J. Biol. Chem. 248:5156

    PubMed  Google Scholar 

  • Bais, R. 1975. A rapid and sensitive radiometric assay for adenosine triphosphatase activity using Cerenkov radiation.Analyt. Biochem. 73:271

    Article  Google Scholar 

  • Bass, A. 1962. Energy metabolism in denervated muscle.In: The Denervated Muscle. E. Gutmann, editor. pp. 175–272. Publishing House of the Czechoslovak Academy of Science, Prague

    Google Scholar 

  • Ben-Bassat, I., Bensch, K.G., Schrier, S.L. 1972. Drug-induced erythrocyte membrane internalization.J. Clin. Invest. 51:1833

    PubMed  Google Scholar 

  • Boegman, R.J., Manery, J.F., Pinteric, L. 1970. The separation and partial purification of membrane bound (Na++K+)dependent Mg2+-ATPase and (Na++K+)-indepent Mg2+-ATP from frog skeletal muscle.Biochim. Biophys. Acta 203:506

    PubMed  Google Scholar 

  • Bray, J.J., Hawken, M.J., Hubbard, J.I. 1974., Prevention by dibutyryl cyclic AMP of denervation-induced fall in muscle fibre resting potential.Proc. Univ. Otago Med. Sch. 52:19

    Google Scholar 

  • Bryant, S.H. 1973. The electrophysiology of myotonia, with a review of congenital myotonia of goats.In: New Developments in Electromyography and Clinical Neurophysiology. J.E. Desmedt, editor. pp. 420–450. Karger, Basel

    Google Scholar 

  • Bryant, S.H., Camerino, D. 1976. Chloride conductance of denervated gastrocnemius fibers from normal goats.J. Neurobiol. 7:229

    Article  PubMed  Google Scholar 

  • Clausen, T., Hansen, O. 1974. Ouabain binding and Na+-K+ transport in rat muscle cells and adipocytes.Biochim. Biophys Acta 345:387

    Google Scholar 

  • Drahota, Z. 1962. Electrolytes in denervated muscle.In: The Denervated Muscle. E. Gutmann, editor. pp. 151–171. Publishing House of the Czechoslovak Academy of Science, Prague

    Google Scholar 

  • Dunham, P.B., Hoffman, J.F. 1970. Partial purification of the ouabain-binding component and of Na, K-ATPase from human red cell membranes.Proc. Nat. Acad. Sci. USA 66:936

    PubMed  Google Scholar 

  • Dunham, P.B., Hoffman, J.F. 1971. Active cation transport and ouabain binding in high potassium and low potassium red blood cells of sheep.J. Gen. Physiol. 58:94

    PubMed  Google Scholar 

  • Fairbanks, G., Steck, T.L., Wallach, D.F.H. 1971. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane.Biochemistry 10:2606

    PubMed  Google Scholar 

  • Festoff, B.W., Engel, W.K. 1974.In vitro analysis of the general properties and junctional receptor characteristics of skeletal muscle membranes. Isolation, purification, and partial characterization of sarcolemmal fragments.Proc. Nat. Acad. Sci. USA 71:2435

    PubMed  Google Scholar 

  • Festoff, B.W., Oliver, K.L., Reddy, N.B. 1977.In vitro Studies of Skeletal Muscle Membranes. Adenylate Cyclase of Fast and Slow Twitch Muscle and the Effects of Denervation.J. Membrane Biol. 32:331

    Google Scholar 

  • Gardner, J.D., Conlon, T.P. 1972. The effects of sodium and potassium on ouabain binding by human erythrocytes.J. Gen. Physiol. 60:609

    PubMed  Google Scholar 

  • Goldberg, A.L., Goodman, H.M. 1968. Effects of disuse and dennervation on amino acid transport by skeletal muscle.Am. J. Physiol. 216:1116

    Google Scholar 

  • Guth, L. 1968. “Tropic” influences of nerve on muscle.Physiol. Rev. 48:645

    PubMed  Google Scholar 

  • Gutmann, E., Zelena, J. 1962. Morphological changes in the denervated muscle.In: The Denervated Muscle. E. Gutmann, editor. pp. 57–102. Publishing House of the Czechoslovak Academy of Science, Prague

    Google Scholar 

  • Lane, L.K., Copenhaver, J.H., Lindenmayer, G.E., Schwartz, A. 1971. Purification and characterization of and3H ouabain binding to the transport adenosine triphosphatase from outer medulla of canine kidney.J. Biol. Chem. 248:7197

    Google Scholar 

  • 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

    PubMed  Google Scholar 

  • Penniston, J.R. 1972. Endocytosis by erythrocyte ghosts: Dependence upon ATP hydrolysis.Arch. Biochem. Biophys. 153:410

    Article  PubMed  Google Scholar 

  • Penniston, J.T., Green, D.E. 1968. The conformational basis of energy transformations in membrane systems. IV. Energized states and pinocytosis in erythrocyte ghosts.Arch. Biochem. Biophys. 128:339

    Article  PubMed  Google Scholar 

  • Reddy, N.B., Engel, W.K., Festoff, B.W. 1976.In Vitro studies of skeletal muscle membranes. Characterization of a phosphorylated intermediate of sarcolemmal (Na++K+) ATPase.Biochim. Biophys. Acta 433:365

    Google Scholar 

  • Rothstein, A. Cabantchik, Z.I., Knauf, P. 1976. Mechanism of anion transport in red blood cells: Role of membrane proteins.Fed. Proc. 35:3

    PubMed  Google Scholar 

  • Schwartz, A., Lindenmayer, G.E., Allen, J.C. 1975. The sodium potassium adenosinetriphosphatase: Pharmacological, physiological, and biochemical aspects.Pharmacol. Rev. 27:3

    PubMed  Google Scholar 

  • Severin, S.E., Boldyrev, A.A., Tkachuk, V.A. 1974. Some properties of Na+, K+-stimulated ATPase from normal and denervated muscles of rabbit with a special reference to its sensitivity to acetylcholine.Comp. Gen. Pharmacol 5:181

    Google Scholar 

  • Taniguchi, K., Iida, S. 1972. Two apparently different ouabain binding sites of Na+, K+-ATPase.Biochim. Biophys. Acta 288:98

    PubMed  Google Scholar 

  • Williams, R.H., Thompson, W.J. 1973. Effect of age upon guanyl cyclase, adenyl cyclase, and cyclic nucleotide phosphodiesterase in rats. Proc. Soc. Exp. Biol. Med.,143:382

    PubMed  Google Scholar 

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Festoff, B.W., Oliver, K.L. & Reddy, N.B. In vitro studies of skeletal muscle membranes. J. Membrain Biol. 32, 345–360 (1977). https://doi.org/10.1007/BF01905227

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  • DOI: https://doi.org/10.1007/BF01905227

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