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OtherCELLULAR AND MOLECULAR PHARMACOLOGY

Effects of Three Sarcoplasmic/Endoplasmic Reticulum Ca++ Pump Inhibitors on Release Channels of Intracellular Stores

Christine Dettbarn and Philip Palade
Journal of Pharmacology and Experimental Therapeutics May 1998, 285 (2) 739-745;
Christine Dettbarn
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Philip Palade
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Abstract

The three principal sarcoplasmic/endoplasmic reticulum Ca++pump inhibitors have been compared for their effects on Ca++ fluxes across intracellular stores present in isolated skeletal muscle and brain membrane preparations. At moderate concentrations that only partially inhibited Ca++ pumping, all three inhibitors induced transient release of Ca++ from isolated sarcoplasmic reticulum membranes, and release was ruthenium red-sensitive, much faster and sustained at higher pump inhibitor concentrations. In contrast, in unidirectional 45Ca efflux assays, cyclopiazonic acid appeared to have little effect, thapsigargin decreased efflux and 2,5-di(tert-butyl)-1,4-benzohydroquinone increased efflux only slightly. These observations taken together suggest that transient releases were manifest primarily by vesicles with a high ratio of ryanodine receptors to pumps (and thus more susceptible to becoming leaky with only some pumps inhibited), and that Ca++-induced Ca++ release amplified releases when all pumps were blocked. These mostly indirect side effects were specific for ryanodine receptors. In similar experiments with brain cerebellar membranes, none of the three inhibitors appeared to directly reduce release induced by inositol 1,4,5-trisphosphate. These findings may affect interpretation of results of experiments involving application of these compounds to isolated membranes, cells or tissue preparations.

Footnotes

  • Send reprint requests to: Dr. Philip Palade, Departments of Physiology and Biophysics and of Pharmacology and Toxicology, University of Texas Medical Branch, Galveston, TX 77555-0641.

  • ↵1 This work was supported by Grant AR43200 from the National Institutes of Health.

  • Abbreviations:
    BSA
    bovine serum albumin
    CICR
    Ca++-induced Ca++ release
    CPA
    cyclopiazonic acid
    di-TBQ
    2′,5′-di(tert-butyl)-1,4-benzohydroquinone
    InsP3
    inositol 1,4,5-trisphosphate
    RR
    ruthenium red
    RyR
    ryanodine receptor
    SCaMPER
    sphingolipid Ca++ release mediating protein of endoplasmic reticulum
    SERCA
    sarcoplasmic/endoplasmic reticulum Ca++ pump
    • Received May 19, 1997.
    • Accepted January 6, 1998.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics
Vol. 285, Issue 2
1 May 1998
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OtherCELLULAR AND MOLECULAR PHARMACOLOGY

Effects of Three Sarcoplasmic/Endoplasmic Reticulum Ca++ Pump Inhibitors on Release Channels of Intracellular Stores

Christine Dettbarn and Philip Palade
Journal of Pharmacology and Experimental Therapeutics May 1, 1998, 285 (2) 739-745;

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OtherCELLULAR AND MOLECULAR PHARMACOLOGY

Effects of Three Sarcoplasmic/Endoplasmic Reticulum Ca++ Pump Inhibitors on Release Channels of Intracellular Stores

Christine Dettbarn and Philip Palade
Journal of Pharmacology and Experimental Therapeutics May 1, 1998, 285 (2) 739-745;
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