Histamine release by compound 48/80: evidence for the depletion and repletion of calcium using chlortetracycline and 45calcium.

Selective release of histamine from rat peritoneal mast cells in vitro was induced by compound 48/80 (1 microgram/ml). Most of the release (75-80%) occurred in a calcium(Ca)-free medium but optimum release was obtained in the presence of 0.9 mM Ca. The release in Ca-free medium still occurred after 180 min incubation. However, prolonged incubation (180 min) in a medium containing chelating agents (EDTA or EGTA) resulted in complete inhibition of histamine release, loss of fluorescence seen with chlortetracycline (CTC) and loss of previously loaded 45Ca from the mast cells. Addition of Ca to these cells resulted in rapid restoration of fluorescence with chlortetracycline. There was also a rapid uptake of 45Ca. Partial depletion of cellular Ca (60 min incubation with EDTA) reduced the rate as well as the amount of histamine release by compound 48/80. These data provide direct evidence for the depletion of cellular Ca which is utilized by compound 48/80 to induce histamine release.