AbstractNoninvasive molecular imaging of subtypes of neuronal nicotinic acetylcholine receptors (nAChRs) will provide information on the role of these receptors in neurodegenerative diseases. The binding of the positron emission tomography ligand 2‐[18F]F‐A‐85380 to nAChRs was investigated in the porcine brain by quantitative autoradiography in vitro. The high‐affinity binding of 2‐[18F]F‐A‐85380 to each of the investigated 12 brain areas was saturable and apparently monophasic (e.g., apparentKDvalue of 1.72 nM in the thalamus). The highest density of specific binding sites was observed in the thalamus (1,158 fmol/mg protein), and the lowest density was measured in the cerebellar gray matter (11 fmol/mg protein). An attempt to assess nAChR subtype specificity of 2‐[18F]F‐A‐85380 was made by competitive autoradiography. Binding of 2‐[18F]F‐A‐85380 coincubated with 2‐F‐A‐85380, epibatidine, cytisine, or methyllycaconitine, respectively, indicated a specificity of 2‐[18F]F‐A‐85380 to β2‐containing nAChRs in the porcine brain. The autoradiographic data confirmed the suitability of swine as a model for the evaluation of radioligands designed for imaging of nAChR subtypes in the living brain. Synapse 59:201–210, 2006. © 2005 Wiley‐Liss, Inc.