HP 749 enhances calcium-independent release of [3H]norepinephrine from rat cortical slices and synaptosomes

Previous studies have shown that, at concentrations of 1 microM and 10 microM, HP 749 increased electrically-stimulated release of [3H]norepinephrine (NE) from rat cortical slices. These effects were Ca(2+)-dependent, indicating an effect on release from vesicular stores. At 100 microM, HP 749 had two effects. In addition to enhancing the Ca(2+)-dependent electrically-evoked release, it also induced a rise in the basal efflux (spontaneous release) of [3H]NE, which was observed in both cortical slices and synaptosomes. The spontaneous release effect was (1) not blocked by the reuptake inhibitor nomifensine, (2) not affected by removal of external calcium, (3) not blocked by vesicular depletion with reserpine, and (4) not inhibited by the sodium channel blocker tetrodotoxin (TTX). As would be expected, the spontaneous [3H]NE release induced by the cytoplasmic releaser tyramine and the sodium channel activator veratridine were blocked by nomifensine and TTX, respectively. Notably, however, the Ca(2+)-independent veratridine-induced release was completely blocked by 100 microM HP 749. The mechanism of spontaneous release of [3H]NE caused by 100 microM HP 749 is unresolved at present; however, the data are consistent with this release originating from a cytoplasmic source.