Ins(1,4,5)P3 and glutathione increase the passive Ca2+ leak in permeabilized A7r5 cells.
Thapsigargin depletes intracellular Ca2+ stores by its inhibitory effect on the Ca2+ pumps, which unmasks an aspecific Ca2+ leak from the stores. This aspecific Ca2+ permeability of the stores was further investigated using 45Ca2+ fluxes on intact and permeabilized A7r5 smooth-muscle cells. Stores in intact cells were found to be more leaky for Ca2+ than those in saponin-permeabilized or Staphylococcus aureus alpha-toxin-permeabilized cells, which suggests that a cytosolic factor may be involved. Supplementing the medium bathing the permeabilized cells with a submaximal Ins(1,4,5)P3 concentration increased the leakiness of the stores. Glutathione also increased the aspecific Ca2+ leak. This effect occurred with both the reduced and the oxidized form but reduced glutathione was more effective. Our data show that basal Ins(1,4,5)P3 levels and glutathione can contribute to the relatively high Ca2+ leak in intact cells. The washing out of these substances during permeabilization can reduce the aspecific leakiness of the stores.
Laboratorium voor Fysiologie, K. U. Leuven, Belgium.
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Parys
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De Smedt
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Casteels
D001427:Bacterial Toxins / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D005978:Glutathione / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D012503:Saponins / Q000494:pharmacology; D013211:Staphylococcus aureus / Q000378:metabolism
D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D012503:Saponins; C021091:staphylococcal alpha-toxin; D015544:Inositol 1,4,5-Trisphosphate; D005978:Glutathione; D002118:Calcium