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OtherPROSTAGLANDINS, LEUKOTRIENES AND OTHER EICOSANOIDS

Mechanism of Action of Leukotriene D4 on Guinea Pig Tracheal Smooth Muscle Cells: Roles of Ca++ Influx and Intracellular Ca++ Release

Daniela Dumitriu, Stéphane Prié, Sylvie G. Bernier, Gaétan Guillemette and Pierre Sirois
Journal of Pharmacology and Experimental Therapeutics March 1997, 280 (3) 1357-1365;
Daniela Dumitriu
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Stéphane Prié
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Sylvie G. Bernier
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Gaétan Guillemette
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Pierre Sirois
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Abstract

The effects of leukotriene D4 (LTD4) on the concentration of intracellular cytosolic free calcium ([Ca++]i) and on phosphoinositide hydrolysis were studied in cultured guinea pig tracheal smooth muscle cells. In Fura-2-loaded cells, LTD4(10−9–10−6 M) induced concentration-dependent changes in [Ca++]iconsisting of a slow, transient increase followed by a sustained phase. Preincubation of cells with LTD4 receptor antagonist MK-571 (10−6 M) blocked the increase in [Ca++]i. Similarly, LTD4-induced inositol phosphate ([3H]InsPs) synthesis was transient, concentration-dependent and inhibited by the LTD4 antagonist. In the absence of extracellular Ca++, LTD4 failed to induce [Ca++]i increases and [3H]InsPs formation. Accordingly, NiCl2 completely inhibited the LTD4-stimulated [3H]InsPs synthesis. Nifedipine (10−5 M) had a slight inhibitory effect on [Ca++]i increase but significantly reduced (40–50%) the [3H]InsPs accumulation. These findings indicate that LTD4-stimulated inositol phosphate synthesis and [Ca++]i increases in tracheal smooth muscle cells are receptor-mediated events and are dependent on the availability of extracellular Ca++. It is suggested that Ca++ influx plays a major role in the LTD4signal transduction mechanism.

Footnotes

  • Send reprint requests to: Pierre Sirois, Ph.D., Department of Pharmacology, Faculty of Medicine, University of Sherbrooke, Sherbrooke, Québec, Canada J1H 5N4.

  • Abbreviations:
    LTD4
    leukotriene D4
    BK
    bradykinin
    [Ca2+]i
    cytosolic calcium ion concentration
    TSMCs
    tracheal smooth muscle cells
    VOC
    voltage-operated calcium channels
    ROC
    receptor-operated calcium channels
    [3H]InsPs
    sum of inositol mono-, bi-, tri- and tetraphosphates
    MK-571
    (3-(((3-(2-(7-chloro-2-quinolinyl) ethenyl) phenyl) ((3-(dimethyl amino-3-oxo propyl)thio)methyl)thio) propanoic acid
    HEPES
    N-[2-hydroxyethyl]piperazine-N′-[2-ethanesulfonic acid]
    DMEM-F12
    Dulbecco’s modified Eagle’s medium and nutrient mixture F-12
    FBS
    fetal bovine serum
    PBS
    phosphate-buffered saline
    Fura-2AM
    Fura-2 pentaacetoxymethyl ester
    EGTA
    ethylene glycol-bis(β-aminoethyl ether)-N, N,N′,N′-tetraacetic acid
    BSA
    bovine serum albumin
    HBSS
    Hanks’ balanced salt solution
    F
    fluorescence
    R
    ratio of the fluorescence at 340 and 380 nm
    PLC
    phospholipase C
    • Received June 21, 1996.
    • Accepted November 25, 1996.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics
Vol. 280, Issue 3
1 Mar 1997
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OtherPROSTAGLANDINS, LEUKOTRIENES AND OTHER EICOSANOIDS

Mechanism of Action of Leukotriene D4 on Guinea Pig Tracheal Smooth Muscle Cells: Roles of Ca++ Influx and Intracellular Ca++ Release

Daniela Dumitriu, Stéphane Prié, Sylvie G. Bernier, Gaétan Guillemette and Pierre Sirois
Journal of Pharmacology and Experimental Therapeutics March 1, 1997, 280 (3) 1357-1365;

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OtherPROSTAGLANDINS, LEUKOTRIENES AND OTHER EICOSANOIDS

Mechanism of Action of Leukotriene D4 on Guinea Pig Tracheal Smooth Muscle Cells: Roles of Ca++ Influx and Intracellular Ca++ Release

Daniela Dumitriu, Stéphane Prié, Sylvie G. Bernier, Gaétan Guillemette and Pierre Sirois
Journal of Pharmacology and Experimental Therapeutics March 1, 1997, 280 (3) 1357-1365;
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