Neuroleptics with differential affinities at dopamine D2 receptors and sigma receptors affect differently the N-methyl-D-aspartate-induced increase in intracellular calcium concentration: involvement of protein kinase.

This study examined the effect of chronic antipsychotic treatment on the NMDA-elicited changes in intracellular free Ca2+ concentration ([Ca2+]i) in the primary culture of rat frontal cortical neurons. Antipsychotics used in the study were chosen for their differential affinities at dopamine D2 receptors and sigma receptors. The potential involvement of protein kinases in this action of antipsychotics were also examined. Chronic treatment of cells with antipsychotics (sulpiride, clozapine, and chlorpromazine) which are known to be potent dopamine D2 receptor ligands, whereas possessing low or no appreciable affinity for sigma receptors, caused a dose-dependent potentiation of the NMDA-induced increase in [Ca2+]i. On the contrary, haloperidol, which is as potent a sigma receptor ligand as a dopamine D2 receptor ligand, did not affect the NMDA-elicited increase in [Ca2+]i. Sulpiride increased the maximum effect afforded by different concentrations of NMDA and shifted the dose-response curve of NMDA to the left (EC50 value from 12.5 microM to 1.39 microM). Consistent with sulpiride's affinity at dopamine D2 receptors, this action of sulpiride was stereoselective: S(-)-sulpiride was active whereas R(+)-sulpiride was inactive. Treatment of cells with dopamine (3 microM) tends to decrease the NMDA-induced increase in [Ca2+]i. Sulpiride at 1 microM totally abolished this action of dopamine and restored its potentiating action on the NMDA-induced increase in [Ca2+]i. Haloperidol, a potent dopamine D2 and sigma receptor ligand, did not affect the sulpiride's potentiating action on the NMDA-induced responses. On the other hand, chronic treatment of cells with a sigma receptor agonist, DTG, at a concentration producing no effect of its own (10 nM), led to an enhancement of the potentiating effect of sulpiride on NMDA-induced increase in [Ca2+]i. This action of DTG was abolished by haloperidol. Further, chronic, but not acute, treatment of cells with either a protein kinase inhibitor H-7 or a cAMP-dependent protein kinase (PKA) inhibitor H-89 abolished this effect of sulpiride on the NMDA-induced [Ca2+]i changes. These results indicate that the action of NMDA in the primary cortical neurons are regulated differently by ligands with differential affinities at dopamine D2 and sigma receptors. The results with protein kinase inhibitors indicate that the potentiation of NMDA responses by sulpiride involves intracellular biochemical events.