clozapine and 1-3-ditolylguanidine

Dopamine D3 receptors may be involved in drug addiction and in disorders such as schizophrenia and Parkinson's disease. To determine the pharmacological properties of dopamine D3 receptors in the rat caudate-putamen, we have investigated R(+)-[3H]7-hydroxy-N,N-di-n-propyl-2-aminotetralin ([3H]R(+)-7-OH-DPAT) binding to membrane preparations from the rat caudate-putamen. Kinetic analyses showed that [3H]R(+)-7-OH-DPAT binding reached equilibrium in approximately 1 h and that both association and dissociation curves were composed of at least two components. Likewise, saturation curves showed at least two binding components with a combined Bmax value of about 600 fmol/mg protein, which is three times higher than what is present in the subcortical limbic area. Competition curves were performed with agonists such as R(-)-propylnorapomorphine, dopamine, PD 128907, quinpirole, and bromocriptine, and antagonists such as haloperidol, raclopride, clozapine, GR 218231x, remoxipride, and U99194A. These experiments revealed that [3H]R(+)-7-OH-DPAT binding could be resolved into three specific binding sites (R1-R3) and one nonspecific binding site, with R1-R2 probably representing D3 receptor binding and the minor R3 representing D2 receptor binding. The low affinities of (+/-)-8-OH-DPAT and 1,3-di(2-tolyl)guanidine to inhibit [3H]R(+)-7-OH-DPAT binding indicate negligible involvement of 5-HT1A or sigma binding sites, respectively. The pharmacological profile of [3H]R(+)-7-OH-DPAT (2 nM) binding in the caudate-putamen was similar to that of dopamine on [125I]iodosulpride binding in the cerebellar lobule X, which contain D3 but not D2 receptors. Mg2+ increased and GTP and Na+ decreased the binding of [3H]R(+)-7-OH-DPAT, suggesting a coupling of endogenous D3 receptors to G proteins. Taken together, these results suggest that dopamine D3 receptors display multiple agonist binding states, and that D3 receptors are present in high concentrations in the rat caudate-putamen. These results may have implications for the physiological and pathological roles of dopamine D3 receptors in the brain.

Topics: 8-Hydroxy-2-(di-n-propylamino)tetralin; Animals; Benzopyrans; Binding, Competitive; Caudate Nucleus; Clozapine; Dopamine; Dopamine Agonists; Dopamine Antagonists; Guanidines; Guanosine Triphosphate; Haloperidol; Heterotrimeric GTP-Binding Proteins; Indans; Kinetics; Magnesium; Male; Oxazines; Protein Binding; Putamen; Quinpirole; Raclopride; Rats; Rats, Sprague-Dawley; Receptors, Dopamine D2; Receptors, Dopamine D3; Remoxipride; Sodium; Specific Pathogen-Free Organisms; Sulfones; Sulpiride; Tetrahydronaphthalenes

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.

Topics: Animals; Antipsychotic Agents; Calcium; Cells, Cultured; Chlorpromazine; Clozapine; Dopamine Antagonists; Frontal Lobe; Guanidines; Haloperidol; N-Methylaspartate; Rats; Rats, Wistar; Receptors, Dopamine D2; Receptors, sigma; Sulpiride