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Nicotine enhances attentional and working memory aspects of executive function in the prefrontal cortex (PFC) where dopamine plays a major role. Here, we have determined the nicotinic acetylcholine receptor (nAChR) subtypes that can modulate dopamine release in rat PFC using subtype‐selective drugs. Nicotine and 5‐Iodo‐A‐85380 (β2* selective) elicited [³H]dopamine release from both PFC and striatal prisms in vitro and dopamine overflow from medial PFC in vivo. Blockade by dihydro‐β‐erythroidine supports the participation of β2* nAChRs. However, insensitivity of nicotine‐evoked [³H]dopamine release to α‐conotoxin‐MII in PFC prisms suggests no involvement of α6β2* nAChRs, in contrast to the striatum, and this distinction is supported by immunoprecipitation of nAChR subunits from these tissues. The α7 nAChR‐selective agonists choline and Compound A also promoted dopamine release from …