The therapeutic and side-effect profiles of clozapine differ from those of typical antipsychotic drugs such as haloperidol. Effects on the serotonin system, especially serotonin-2 receptors, may contribute to clozapine's atypicality. We injected rats for 14 days with clozapine (25 mg/kg/day) or haloperidol (2 mg/kg/day), and measured three aspects of the serotonin system in forebrain regions: abundance of serotonin-2A, -2C and -1A receptor messenger RNAs by in situ hybridization histochemistry; serotonin-2A and -1A binding sites using receptor autoradiography, and levels of serotonin and 5-hydroxyindoleacetic acid with high-performance liquid chromatography. Clozapine administration decreased serotonin-2A receptor messenger RNA and the density of [3H]ketanserin binding in cingulate and frontal cortex, but not in piriform cortex. Serotonin-1A receptor expression and serotonin-2C receptor messenger RNA were unchanged in all areas. The treatment markedly decreased serotonin and 5-hydroxyindoleacetic acid concentrations in striatum with similar trends in cortex and hippocampus. Haloperidol administration did not affect the expression of the three serotonin receptors, but was associated with a modest reduction of striatal and hippocampal 5-hydroxyindoleacetic acid. The selective reduction of serotonin-2A receptors confirms earlier findings and supports the view that this receptor may have relevance for the actions of clozapine. The fact that the encoding messenger RNA is decreased shows that the the effect is mediated at the level of gene expression. In contrast, the unchanged serotonin-2C receptor messenger RNA level indicates that the reported loss of serotonin-2C receptors after clozapine treatment is due to translational or post-translational events. The relationship between the reduction in serotonin-2A receptor expression and the altered serotonin metabolism remains unclear.