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    Refilling of caffeine-sensitive intracellular calcium stores in bovine airway smooth muscle cells

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    Authors
    Madison, John M.
    Ethier, Michael F.
    Yamaguchi, Hiroshi
    UMass Chan Affiliations
    Departments of Medicine and Physiology
    Document Type
    Journal Article
    Publication Date
    1998-11-14
    Keywords
    Animals
    Caffeine
    Calcium
    Cattle
    Cells, Cultured
    Fluorescent Dyes
    Fura-2
    Genistein
    Kinetics
    Microscopy, Fluorescence
    Muscle, Smooth
    Nickel
    Trachea
    Life Sciences
    Medicine and Health Sciences
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    Link to Full Text
    https://doi.org/10.1152/ajplung.1998.275.5.L852
    Abstract
    The goal of this study was to assess the mechanisms by which the caffeine-sensitive calcium stores of airway smooth muscle cells are refilled. Bovine trachealis cells were loaded with fura 2-AM (0.5 microM) for imaging of cytosolic calcium concentrations ([Ca2+]i) in the inner cytosol. After a first stimulation (S1) with caffeine, the response to a second stimulation (S2) depended on the presence of extracellular calcium during an intervening 80-s-long refilling phase. The S2-to-S1 ratio (S2/S1) was 0.11 +/- 0.05 (n = 13 cells) during calcium-free refilling but 0.72 +/- 0.04 (n = 36 cells) within 80 s of exposure to extracellular calcium. Maximum mean [Ca2+]i during the 80 s of refilling was not different for calcium-free (116 +/- 19 nM; n = 13 cells) versus extracellular calcium plus nickel (2 mM) (121 +/- 12 nM; n = 21 cells); despite this, significantly greater refilling (S2/S1 0.58 +/- 0.06; n = 24 cells) occurred in the presence of extracellular calcium plus nickel. The protein tyrosine kinase inhibitors genistein (100 microM) and ST-638 (50 microM) significantly decreased refilling over 80 s (S2/S1 0.35 +/- 0.06, n = 14 cells and 0.51 +/- 0.07, n = 14 cells, respectively). Daidzein (100 microM) had no effect on S2/S1. We concluded that [Ca2+]i of the inner cytosol during refilling correlated poorly with S2/S1 values and that, therefore, additional compartments not well detected by fura 2 contribute to refilling. The findings suggest that calcium influx for refilling is segregated from the inner cytosol of the cell, relatively insensitive to nickel, and regulated or modulated by protein tyrosine kinase activity.
    Source

    Am J Physiol. 1998 Nov;275(5 Pt 1):L852-60.

    DOI
    10.1152/ajplung.1998.275.5.L852
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/38189
    PubMed ID
    9815101
    Related Resources

    Link to article in PubMed

    ae974a485f413a2113503eed53cd6c53
    10.1152/ajplung.1998.275.5.L852
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