Mechanism of the dopamine-releasing actions of amphetamine and cocaine: plasmalemmal dopamine transporter versus vesicular monoamine transporter.

The effects of amphetamine and cocaine were studied in [3H]-dopamine-loaded and superfused COS-7 cells transfected with either the cDNA of the plasmalemmal dopamine transporter ("DAT cells") or the cDNA of the vesicular amine transporter ("VAT cells"), or with both transporters ("DAT/VAT cells"). Amphetamine (0.01-100 microM, added for 4 min of superfusion) led to a concentration-dependent increase in dopamine release in DAT cells, as well as in DAT/VAT cells. The EC50 of the effect of amphetamine on DAT cells was 1.1 +/- 0.6 microM; the effect on DAT/VAT cells did not reach a plateau in the concentration range tested. With longer exposure to amphetamine, dopamine efflux from DAT cells reached a peak and quickly returned to baseline, in spite of the continued presence of the drug, whereas in DAT/VAT cells and in VAT cells the effect was sustained. Cocaine (up to 100 microM) did not exert any effect of its own in DAT cells or VAT cells but inhibited the amphetamine-induced release of dopamine from DAT cells in a competitive manner. In DAT/VAT cells cocaine and its analogue (-)-2 beta-carbomethoxy-3 beta-(4-fluorophenyl)tropane caused an efflux of dopamine resembling that caused by amphetamine but quantitatively much smaller. The rank order of potency was the same as in uptake experiments [(-)-2 beta-carbomethoxy-3 beta-(4-fluorophenyl)tropane > cocaine]. The effect of cocaine was mimicked by the reduction of chloride. The results indicate that there is a plasmalemmal component and a vesicular component in the dopamine-releasing action of amphetamine. The releasing action of cocaine is dependent on the existence of a vesicular pool of the neurotransmitter and seems to be linked to inhibition of the plasmalemmal dopamine transporter.