Skip to main content
SpringerLink
Log in

Skeletal muscle ATP-sensitive K+ channels recorded from sarcolemmal blebs of split fibers: ATP inhibition is reduced by magnesium and ADP

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

A new, nonenzymatically treated preparation of amphibian sarcolemmal blebs has been used to study the regulation of skeletal muscle ATP-sensitive K+ [K(ATP)] channels.

When a frog skeletal muscle fiber is split in half in a Ca2+-free relaxing solution, large hemispherical membrane blebs appear spontaneously within minutes without need for Ca2+-induced contraction or enzymatic treatment. These blebs readily formed gigaseals with patch pipettes, and excised inside-out patches were found to contain a variety of K+ channels. Most prominent were K(ATP) channels similar to those found in the surface membrane of other muscle and nonmuscle cells. These channels were highly selective for K+, had a conductance of ≈ 53 pS in 140mmK+, and were blocked by internal ATP. The presence of these channels in most patches implies that split-fiber blebs are made up, at least in large part, of sarcolemmal membrane.

In this preparation, K(ATP) channels could be rapidly and reversibly blocked by glibenclamide (0.1–10 μm) in a dose-dependent manner. These channels were sensitive to ATP in the micromolar range in the absence of Mg. This sensitivity was noticeably reduced in the presence of millimolar Mg, most likely because of the ability of Mg2+ ions to bind ATP. Our data therefore suggest that free ATP is a much more potent inhibitor of these channels than MgATP. Channel sensitivity to ATP was significantly reduced by ADP in a manner consistent with a competition between ADP, a weak inhibitor, and ATP, a strong inhibitor, for the same inhibitory binding sites.

These observations suggest that the mechanisms of nucleotide regulation of skeletal muscle and pancreatic K(ATP) channels are more analogous than previously thought.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Arena, J.P., Kass, R.S. 1989. Activation of ATP-sensitive K channels in heart cells by pinacidil: Dependence on ATP.Am. J. Physiol. 257:H2092-H2096

    Google Scholar 

  • Ashcroft, F.M., Kakei, M. 1989. ATP-sensitive K− channels in pancreatic B-cells: Modulation by ATP and Mg2+ ions.J. Physiol. 416:349–367

    Google Scholar 

  • Ashcroft, S.J.H., Ashcroft, F.M. 1990. Properties and functions of ATP-sensitive K-channels.Cell. Signal. 2:197–214

    Google Scholar 

  • Belles, B., Hescheler, J., Trube, G. 1987. Changes of membrane currents in cardiac cells induced by long whole-cell recordings and tolbutamide.Pfluegers Arch. 409:582–588

    Google Scholar 

  • Burton, F., Dörstelmann, U., Hutter, O.F. 1988. Single-channel activity in sarcolemmal vesicles from human and other mammalian muscles.Muscle Nerve 11:1029–1038

    Google Scholar 

  • Davies, N.W. 1990. Modulation of ATP-sensitive K+ channels in skeletal muscle by intracellular protons.Nature 343:375–377

    Google Scholar 

  • Dunne, M.J., Illot, M.C., Petersen, O.H. 1987. Interaction of diazoxyde, tolbutamide and ATP4− on nucleotide-dependent K+ channels in an insulin-secreting cell line.J. Membrane Biol. 99:215–224

    Google Scholar 

  • Dunne, M.J., Petersen, O.H. 1986. Intracellular ADP activates K+ channels that are inhibited by ATP in an insulin-secreting cell line.FEBS Lett. 208:59–62

    Google Scholar 

  • Dunne, M.J., West-Jordan, J.A., Abraham, R.J., Edwards, R.H.T., Petersen, O.H. 1988. The gating of nucleotide-sensitive K+ channels in insulin-secreting cells can be modulated by changes in the ratio ATP4−/ADP3− and by nonhydrolyzable derivatives of both ATP and ADP.J. Membrane Biol. 104:165–172

    Google Scholar 

  • Fabiato, A. 1988. Computer programs for calculating total from specified or free from specified total ionic concentrations in aqueous solutions containing multiple metals and ligands.In: Methods in Enzymology. Vol. 157. pp. 378–417. S. Fleischer and B. Fleischer, editors. Academic, San Diego

    Google Scholar 

  • Fan, Z., Nakayama, K., Hiraoka, M. 1990. Pinacidil activates the ATP-sensitive K+ channel in inside-out and cell-attached patch membranes of guinea-pig ventricular myocytes.Pfluegers Arch. 415:387–394

    Google Scholar 

  • Findlay, I. 1987a. ATP-sensitive K+ channels in rat ventricular myocytes are blocked and inactivated by internal divalent cations.Pfluegers Arch. 410:313–320

    Google Scholar 

  • Findlay, I. 1987b. The effects of magnesium upon adenosine triphosphate-sensitive potassium channels in a rat insulinsecreting cell line.J. Physiol. 391:611–629

    Google Scholar 

  • Findlay, I. 1988a. ATP4− and ATP·Mg inhibit the ATP-sensitive K+ channel of rat ventricular myocytes.Pfluegers Arch. 412:37–41

    Google Scholar 

  • Findlay, I. 1988b. Effects of ADP upon the ATP-sensitive K+ channel in rat ventricular myocytes.J. Membrane Biol. 101:83–92

    Google Scholar 

  • Findlay, I., Dunne, M.J. 1986. ATP maintains ATP-inhibited K+ channels in an operational state.Pfluegers Arch. 407:238–240

    Google Scholar 

  • Fosset, M., de Weille, J., Green, R.D., Schmid-Antomarchi, H., Lazdunski, M. 1988. Antidiabetic sulfonylureas control action potential properties in heart cells via high-affinity receptors that are linked to ATP-dependent K+ channels.J. Biol. Chem. 263:7933–7936

    Google Scholar 

  • Gillis, K.D., Gee, W.M., Hammoud, A., McDaniel, M.L., Falke, L.C., Misler, S. 1989. Effects of sulfonamides on a metaboliteregulated ATP-sensitive K+ channel in rat pancreatic B-cells.Am. J. Physiol. 257:C1119-C1127

    Google Scholar 

  • Grafe, P., Quasthoff, S., Strupp, M., Lehmann-Horn, F. 1990. Enhancement of K+ conductance improves in vitro the contraction force of skeletal muscle in hypokalemic periodic paralysis.Muscle Nerve 13:451–457

    Google Scholar 

  • Hamill, O.P., Marty, A., Neher, E., Sakman, B., Sigworth, F.J. 1981. Improved patch-clamp techniques for high-resolution current recordings from cells and cell-free membrane patches.Pfluegers Arch. 391:85–100

    Google Scholar 

  • Hilgemann, D.W. 1989. Giant excised cardiac sarcolemmal membrane patches: Sodium and sodium-calcium exchange currents.Pfluegers Arch. 415:247–249

    Google Scholar 

  • Horie, M., Irisawa, H., Noma, A. 1987. Voltage-dependent magnesium block of adenosine-triphosphate-sensitive potassium channel in guinea-pig ventricular cells.J. Physiol. 387:251–272

    Google Scholar 

  • Ishikawa, H., Sawada, H., Yamada, E. 1983. Surface and internal morphology of skeletal muscle.In: Handbook of Physiology. Sect. 10: Skeletal Muscle. L.H. Peachey, editor. pp. 1–21. Williams & Wilkins, Baltimore

    Google Scholar 

  • Kakei, M., Kelly, R.P., Ashcroft, S.J.H., Ashcroft, F.M. 1986. The ATP-sensitivity of K+ channels in rat pancreatic B-cells is modulated by ADP.FEBS Lett. 208:63–66

    Google Scholar 

  • Latorre, R., Oberhauser, A., Labarca, P., Alvarez, O. 1989. Varieties of calcium-activated potassium channels.Annu. Rev. Physiol. 51:385–399

    Google Scholar 

  • Lederer, J.W., Nichols, C.G. 1989. Nucleotide regulation of the activity of rat heart ATP-sensitive K− channels in isolated membrane patches.J. Physiol. 419:193–211

    Google Scholar 

  • Misler, S., Falke, L.C., Gillis, K., McDaniel, M.L. 1986. A metabolite-regulated potassium channel in rat pancreatic B cells.Proc. Natl. Acad. Sci. USA 83:7119–7123

    Google Scholar 

  • Mitchell, R.D., Palade, P., Fleischer, S. 1983. Purification of morphologically intact triad structures from skeletal muscle.J. Cell. Biol. 96:1008–1016

    Google Scholar 

  • Parent, L., Coronado, R. 1989. Reconstitution of the ATP-sensitive potassium channel of skeletal muscle.J. Gen. Physiol. 94:445–463

    Google Scholar 

  • Qin, D., Takano, M., Noma, A. 1989. Kinetics of ATP-sensitive K− channel revealed with oil-gate concentration jump method.Am. J. Physiol. 257:H1264–1633

    Google Scholar 

  • Quast, U., Cook, N.S. 1989. Moving together: K+ channel openers and ATP-sensitive K+ channels.Trends Pharmacol. Sci. 10:431–435

    Google Scholar 

  • Quasthoff, S., Franke, C., Hatt, H., Richter-Turtur, M. 1990. Two different types of potassium channels in human skeletal muscle activated by potassium channel openers.Neurosci. Lett. 119:191–194

    Google Scholar 

  • Schmid-Antomarchi, H., de Weille, J., Fosset, M., Lazdunski, M. 1987. The antidiabetic sulfonylurea glibenclamide is a potent blocker of the ATP-modulated K+ channel in insulin secreting cells.Biochem. Biophys. Res. Commun. 146:21–25

    Google Scholar 

  • Spruce, A.E., Standen, N.B., Stanfield, P.R. 1985. Voltage-dependent ATP-sensitive potassium channels of skeletal muscle membrane.Nature 316:736–738

    Google Scholar 

  • Spruce, A.E., Standen, N.B., Stanfield, P.R. 1987. Studies of the unitary properties of adenosine-5′-triphosphate-regulated potassium channels of frog skeletal muscle.J. Physiol. 382:213–236

    Google Scholar 

  • Spuler, A., Lehmann-Horn, F., Grafe, P. 1989. Cromakalim (BRL 34915) restores in vitro the membrane potential of depolarized human skeletal muscle fibres.Naunyn Schmiedebergs Arch. Pharmacol. 339:327–331

    Google Scholar 

  • Standen, N.B., Quayle, J.M., Davies, N.W., Brayden, J.E., Huang, Y., Nelson, M.T. 1989. Hyperpolarizing vasodilators activate ATP-sensitive K+ channels in arterial smooth muscle.Science 245:177–180

    Google Scholar 

  • Standen, N.B., Stanfield, P.R., Ward, T.A., Wilson, S.W. 1984. A new preparation for recording single-channel currents from skeletal muscle.Proc. R. Soc. London B. 221:455–464

    Google Scholar 

  • Stein, P., Palade, P. 1988.Sarcoballs: Direct access to sarcoplasmic reticulum Ca2+ channels in skinned frog muscle fibers.Biophys. J. 54:357–363

    Google Scholar 

  • Stein, P., Palade, P. 1989. Patch clamp of sarcolemmal spheres from stretched skeletal muscle fibers.Am. J. Physiol. 256:C434-C440

    Google Scholar 

  • Sturgess, N.C., Ashford, M.L.J., Cook, D.L., Hales, C.N. 1985. The sulphonylurea receptor may be an ATP-sensitive potassium channel.Lancet 8453:474–475

    Google Scholar 

  • Trube, G. 1983. Enzymatic dispersion of heart and other tissues.In: Single-Channel Recordings. B. Sakmann and E. Neher, editors. pp. 69–76. Plenum, New York

    Google Scholar 

  • Vivaudou, M.B., Arnoult, C., Villaz, M. 1989. Patch-clamp recordings cordings of K+ channels from membrane blebs (‘Sarcoballs’) from skinned frog skeletal muscle fibers.Biophys. J. 55:483a

    Google Scholar 

  • Vivaudou, M.B., Arnoult, C., Villaz, M. 1990. Skeletal muscle ATP-sensitive K+ channels recorded from sarcolemmal vesicles obtained by a new non-enzymatic method.Biophys. J. 57:512a

    Google Scholar 

  • Vivaudou, M.B., Robert, M., Arnoult, C., Villaz, M. 1990. Skeletal muscle ATP-sensitive K+ channels recorded from sarcolemmal vesicles of split fibers: Regulation by internal ATP and ADP.J. Neurol. Sci. 98(Suppl.):221

    Google Scholar 

  • Weik, R., Lönnendonker, U., Neumcke, B. 1989. Low-conductance states of K+ channels in adult mouse skeletal muscle.Biochim. Biophys. Acta 983:127–134

    Google Scholar 

  • Weik, R., Neumcke, B. 1989. ATP-sensitive potassium channels in adult mouse skeletal muscle: Characterization of the ATP-binding site.J. Membrane Biol. 110:217–226

    Google Scholar 

  • Woll, K.H., Lönnendonker, U., Neumcke, B. 1989. ATP-sensitive potassium channels in adult mouse skeletal muscle: Different modes of blockage by internal cations, ATP and tolbutamide.Pfluegers Arch. 414:622–628

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Biophysique Moléculaire et Cellulaire (CNRS UA520) BMC/DBMS/CENG, 85X, 38041, Grenoble, France

    Michel B. Vivaudou, Christophe Arnoult & Michel Villaz

Authors
  1. Michel B. Vivaudou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christophe Arnoult
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michel Villaz
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vivaudou, M.B., Arnoult, C. & Villaz, M. Skeletal muscle ATP-sensitive K+ channels recorded from sarcolemmal blebs of split fibers: ATP inhibition is reduced by magnesium and ADP. J. Membrain Biol. 122, 165–175 (1991). https://doi.org/10.1007/BF01872639

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01872639

Key Words

Search

Navigation