AbstractWe have recently clarified that nicotinic acetylcholine receptors (nAChRs) expressed in the thalamus play an important role in antiallodynic effects produced by the nAChR agonist, 5‐iodo‐3‐(2(S)‐azetidinylmethoxy)pyridine (5IA). This study aimed to reveal the in vivo relationship between thalamic nAChR occupancy rates and antiallodynic effects using 5IA and [125I]5IA. We partially ligated the sciatic nerve of a rat to induce neuropathic pain. Antiallodynic effects were evaluated at 15, 30, 60, and 90 min after intracerebroventricular (i.c.v.) administration of multiple doses (1–100 nmol) of 5IA by the von Frey filament test. Receptor occupancy rates were measured by autoradiography at 15 and 90 min after administration. Antiallodynic effects of repetitive treatment of 5IA (5 and 50 nmol) were also examined. A significant and dose‐dependent antiallodynic effect was observed 15 min after administration. It showed a good correlation with receptor occupancy rates (r= 0.97), indicating the binding of 5IA to nAChRs expressed in the thalamus involved in the antiallodynic effect. Five, 50, and 100 nmol of 5IA occupied the thalamic nAChRs until 90 min after administration, while the antiallodynic effect diminished. Five nanomoles of 5IA (which occupied 40% of thalamic nAChRs) showed a significant antiallodynic effect (percentage of the maximal possible effect (%MPE): 35 ± 7) after the second administration, while 50 nmol of 5IA (which occupied 80% of thalamic nAChRs) did not (%MPE: 7 ± 1). These findings suggest that not clearance of 5IA but desensitization of nAChRs caused by persistent binding of 5IA is responsible for the disappearance of antiallodynic effects. Synapse 65:77–83, 2011. © 2010 Wiley‐Liss, Inc.