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The prototypical neuroleptic haloperidol and the atypical antipsychotic clozapine induce distinctly different patterns of c-fos expression in the forebrain. While haloperidol appears to increase c-fos expression via its D2 dopamine receptor antagonist properties, the receptor mechanisms by which clozapine produces its unique pattern of c-fos expression are not known. The present experiments sought to address this question by determining the phenotypes of neurons in which clozapine increases Fos-like immunoreactivity (FLI). Fos immunostaining combined with in situ hybridization histochemistry using a cDNA oligonucleotide probe for D3 receptor mRNA indicated that the great majority (95%) of clozapine-induced FLI neurons in the major island of Calleja (ICjM) express D3 receptors. Similarly, in the nucleus accumbens (NAc) and lateral septal nucleus (LSN), the majority of clozapine-induced FLI neurons express D3 receptor mRNA (NAc 69%; LS 73%). In marked contrast, haloperidol-induced FLI neurons failed to express D3 receptors in any brain region. Studies with oligonucleotide probes for enkephalin (ENK) and dynorphin (DYN) indicated that clozapine increases c-fos expression in both ENK and DYN containing neurons in the NAc (ENK 40%, DYN 53%) and LSN (ENK 32%, DYN 59%). Haloperidol also increases c-fos expression in ENK and DYN containing neurons, albeit in a different pattern (striatum: ENK 93%, DYN 20%; nucleus accumbens: ENK 46%, DYN 36%; lateral septum: ENK 29%, DYN 18%). The present results demonstrate that haloperidol and clozapine target different populations of neurons even in regions such as the NAc and LSN, where they both increase c-fos expression. In addition, the fact that the majority of clozapine-sensitive neurons in NAc, LSN, and ICjM express D3 receptors suggests that activity at these receptors may contribute to the unique clinical profile of this antipsychotic agent. These data indicate that D3 receptors may represent novel targets in the pharmacotherapy of schizophrenia.

Phenotypic Characterization of Neuroleptic-Sensitive Neurons in the Forebrain Contrasting Targets of Haloperidol and Clozapine