Our previous studies have shown several fold induction of neurotensin/neuromedin N (NT/N) primary transcripts and mature cytosolic mRNA in the dorsolateral region of the rat neostriatum (DLSt) following a single dose of the neuroleptic, haloperidol. The apparent enhancement of NT/N gene transcription by haloperidol is preceded by increases in the expression of c-fos mRNA in the same region. The present study used double-labeling in situ hybridization technique to study cellular localization of NT/N and c-fos mRNA following acute haloperidol treatment. Simultaneous detection of NT mRNA and c-fos mRNA was achieved using cRNA probes synthesized in vitro with digoxigenin- and 35S-labeled UTP, respectively. Interestingly, approximately 75% of DLSt neurons expressing NT/N mRNA also displayed c-fos mRNA in rats treated with haloperidol (1 mg/kg, i.p.) for 1 h. Colocalization of c-fos mRNA in haloperidol-responsive NT neurons in the DLSt suggests that haloperidol's induction of NT/N gene transcription may involve participation of the transcription factor Fos and the AP-1 consensus sequence in regulatory region of the NT/N gene.