Beta2* Nicotinic Receptors Tune Prefrontal Behavior Through Distinct Neuronal Populations

Beta2 subunit-containing nicotinic acetylcholine receptors (beta2* nAChRs) are highly expressed in the prefrontal cortex (PFC) and critically regulate behavioral and cognitive domains disrupted in neuropsychiatric disorders. Despite their therapeutic potential, the cell-type- and circuit-specific functions of beta2* nAChRs remain poorly defined, partially due to the lack of selective pharmacological tools. Here, we delineate the cellular expression patterns and behavioral roles of beta2* nAChRs across distinct neuronal populations of the mouse PFC. Using fluorescence in situ hybridization (FISH), we mapped beta2 nicotinic subunit expression across cortical layers and major neuronal classes. We then employed CRISPR-mediated knockdown (KD) to selectively reduce beta2* nAChRs expression in two distinct PFC neuronal populations and, for comparison, in a striatal population. Selective KD of beta2* nAChRs in deep-layer PFC neurons defined by early expression of neuropeptide Y (NPY) disrupted working memory and exploratory behavior and reshaped social and anxiety-like behaviors. In contrast, KD in superficial-layer PFC interneurons expressing the serotonin receptor 5HT3a induced a robust hypersocial phenotype, accompanied by exploratory behavior changes overlapping with those observed following NPY KD. Notably, KD of beta2* nAChRs in a rare NPY-expressing striatal population affected similar behavioral domains, but largely in the opposite direction to PFC manipulations. Together, these findings reveal opposing, cell-type- and region-specific contributions of beta2* nAChRs to behavioral regulations, highlighting the necessity of circuit-resolved targeting strategies to improve the precision and efficacy of therapeutic intervention for neuropsychiatric disorders.