Comparison of the effect of cyclic AMP on the content and release of dopamine and 1‐methyl‐4‐phenylpyridinium (MPP+) in PC12 cells

1 The aim of this work was to investigate the effect of acute and chronic exposure of rat pheochromocytoma (PC12) cells to elevated cAMP, using forskolin, dibutyryl-cAMP (db-cAMP) or isobutylmethylxanthine (IBMX), on endogenous dopamine content and release and on [3H]-1-methyl-4-phenylpyridinium ([3H]-MPP+) uptake and release, under basal conditions and under KCl-stimulation. 2 Cultured PC12 cells synthetized and accumulated large amounts of dopamine, but not noradrenaline or adrenaline. The release of dopamine by the cells was markedly increased in response to 50 mM KCl. 3 Acute and chronic treatment of the cells with forskolin (30 microM), but not IBMX (100 microM), slightly increased the spontaneous release of dopamine and significantly decreased the release induced by 50 mM KCl. 4 Chronic treatment of the cells with forskolin (30 microM), but not IBMX (100 microM), markedly decreased the cellular content of dopamine. 5 Cultured PC12 cells removed and accumulated [3H]-MPP+, which, similarly to dopamine, was released by KCl. 6 Acute treatment of the cells with forskolin (30 microM) or db-cAMP (2.5 mM), but not IBMX (100 microM), slightly increased the spontaneous release, but did not affect KCl-induced release of [3H]-MPP+. On the other hand, chronic treatment of the cells with forskolin produced, on [3H]-MPP+, similar effects to those obtained for dopamine. 7 Acute and chronic treatment of the cells with reserpine (50 nM) produced similar results to those obtained with forskolin on either dopamine or [3H]-MPP+ handling. 8 In conclusion, cAMP, similarly to reserpine, increases the spontaneous release and decreases the KCl-induced release of [3H]-MPP+ and dopamine. This suggests that cAMP impairs the vesicular monoamine transporter.