Ecstasy and methamphetamine elicit action potential bursts via different mechanisms in a central snail neuron

This study sought to determine the effects of (+) methamphetamine (METH) and its ring-substituted analog (±)3,4-methylenedioxymethamphetamine (MDMA; ecstasy) on electrophysiological behavior and their relationships to second messenger systems in an identifiable RP4 neuron of the African snail, Achatina fulica Ferussac. Extracellular application of MDMA at 1 mM and METH at 3 mM elicited action potential bursts that were not blocked after immersing the neurons in Ca2+-free solution. Notably, MDMA- (1 mM) elicited action potential bursts were blocked by pretreatment with the protein kinase C (PKC) inhibitors chelerythrine (20 μM) and Ro 31-8220 (20 μM), but not by the PKA inhibitors KT-5720 (10 μM) and H89 (10 μM). The PKC activator phorbol 12,13-dibutyrate (PDBu; 3 μM), but not the PKA activator forskolin (50 μM), facilitated the induction of bursts elicited by MDMA at a lower concentration (0.3 mM). In contrast, METH- (3 mM) elicited action potential bursts were blocked by pretreatment with KT-5720 (10 μM) and H89 (10 μM), but not by chelerythrine (20 μM) and Ro 31-8220 (20 μM). Forskolin (50 μM), but not PDBu (3 μM) facilitated the induction of bursts elicited by METH at a lower concentration (1 mM). Tetraethylammonium chloride (TEA), a blocker of the delayed rectifying K+ current (IKD), did not elicit bursts at a concentration of 5 mM but did facilitate the induction of action potential bursts elicited by both METH and MDMA. Voltage clamp studies revealed that both METH and MDMA decreased the TEA-sensitive IKD of the RP4 neuron. Forskolin (50 μM) or dibutyryl cAMP (1 mM), a membrane-permeable cAMP analog, alone did not elicit action potential bursts. However, co-administration with forskolin (50 μM) and TEA (5 mM) or co-administration with dibutyryl cAMP (1 mM) and TEA (50 mM) elicited action potential bursts in the presence of the PKC inhibitor chelerythrine (20 μM). Similarly, PDBu (10 μM) or phorbol 12-myristate 13-acetate (PMA; 3 μM) alone did not elicit action potential bursts. However, co-administration with PDBu (10 μM) and TEA (5 mM) or co-administration with PMA (3 μM) and TEA (5 mM) elicited action potential bursts in the presence of the PKA inhibitor KT-5720 (10 μM). These data suggest that action potential bursts in the RP4 neuron were not due to Ca2+-dependent synaptic effects. Rather, action potential bursts may be elicited through (1) combined activation of the cAMP-PKA signaling pathway and inhibition of the IKD and (2) combined activation of PKC and inhibition of the IKD.