Studies on histamine-retaining granules obtained from isolated rat mast cells

Histamine-retaining granules were isolated from rat mast cells after sonication in either sucrose of Ficoll-Hypaque media. The preparations obtained were compared in regard to recovery and spontaneous loss of histamine. The effect of agents known to release histamine from intact rat mast cells (antigen, compound 48/80, decylamine, the ionophores A23187 and X537A as well as ATP) was studied on the granules. Antigen and compound 48/80 did not release histamine. Decylamine and X537A induced a pronounced release independent of the presence of divalent cations. ATP caused a small, but significant release, which showed an absolute requirement for magnesium. A23187 released histamine only in the presence of either calcium or magnesium, and this release was unaffected by certain agents known to inhibit histamine release from intact rat mast cells. The results seem to exclude the possibility that agents known to induce release of histamine from intact rat mast cells by a calcium-and energy-dependent process would exert this action through a direct effect on intracellularly localized granules.