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Blockade of insulin sensitive steady-state R-type Ca2+ channel by PN 200-110 in heart and vascular smooth muscle

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Abstract

The effect of high K− concentration, insulin and the L-type Ca2− channel blocker PN 200-110 on cytosolic intracellular free calcium ([Ca2+]i) was studied in single ventricular myocytes of 10-day-old embryonic chick heart, 20-week-old human fetus and rabbit aorta (VSM) single cells using the Ca2+-sensitive fluorescent dye, Fura-2 microfluorometry and digital imaging technique. Depolarization of the cell membrane of both heart and VSM cells with continuous superfusion of 30 mM [K+]o induced a rapid transient increase of [Ca2+]i that was followed by a sustained component. The early transient increase of [Ca2+]i by high [+]o was blocked by the L-type calcium channel antagonist nifedipine. However, the sustained component was found to be insensitive to this drug. PN 200-110 another L-type Ca2+ blocker was found to decrease both the early transient and the sustained increase of [Ca2+]i induced by depolarization of the cell membrane with high [K+]o. Insulin at a concentration of 40 to 80 μU/ml only produced a sustained increase of [Ca2+]i that was blocked by PN 200-110 or by lowering the extracellular Ca2+ concentration with EGTA. The sustained increase of [Ca2+], induced by high [K+]o or insulin was insensitive to metabolic inhibitors such as KCN and ouabain as well to the fast Na+ channel blocker, tetrodotoxin and to the increase of intracellular concentrations of cyclic nucleotides. Using the patch clamp technique, insulin did not affect the L-type Ca2+ current and the delayed outward K+ current. These results suggest that the early increase of (Ca2+]i during depolarization of the cell membrane of heart and VSM cells with high [K+]o is due to the opening and decay of an L-type Ca 2+ channel. However, the sustained increase of [Ca2+]i during a sustained depolarization is due to the activation of a resting (R) Ca 2+ channel that is insensitive to lowering [ATP]i and sensitive to insulin.

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Authors and Affiliations

  1. Dept of Physiology and Biophysics, Faculty of Medicine, University of Sherbrooke, JIH 5N4, Sherbrooke, Quebec, Canada

    G. Bkaily, D. Econornos, L. Potvin & C. Marriott

  2. Dept of Endocrinology, Faculty of Medicine, University of Sherbrooke, JIH 5N4, Sherbrooke, Quebec, Canada

    J. -L. Ardilouze

  3. Dept of Urology, Faculty of Medicine, University of Sherbrooke, JIH 5N4, Sherbrooke, Quebec, Canada

    J. Corcos

  4. Dept of Cardiology, Faculty of Medicine, University of Sherbrooke, JIH 5N4, Sherbrooke, Quebec, Canada

    D. Bonneau

  5. Department of Scientific Development, Sandoz Canada, 385 Bouchard Blvd, H9R 4P5, Dorval, Quebec, Canada

    Chin Nok Fong

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  1. G. Bkaily
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  2. D. Econornos
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  3. L. Potvin
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  4. J. -L. Ardilouze
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  5. C. Marriott
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  6. J. Corcos
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  7. D. Bonneau
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  8. Chin Nok Fong
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Bkaily, G., Econornos, D., Potvin, L. et al. Blockade of insulin sensitive steady-state R-type Ca2+ channel by PN 200-110 in heart and vascular smooth muscle. Mol Cell Biochem 117, 93–106 (1992). https://doi.org/10.1007/BF00230415

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  • DOI: https://doi.org/10.1007/BF00230415

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