The lateral olfactory tract (LOT) is a central olfactory pathway, and efferent projections from the olfactory bulb are conveyed to the olfactory-related cortical structures via the LOT. The purpose of the present study is to determine the exact site of the LOT causing functional impairment in animals. After ablation of the right olfactory bulb, rats received rostrocaudal transection injuries on the left LOT at different levels between the olfactory bulb and the middle cerebral artery. Olfactory function of LOT-transected rats was studied by examining their olfactory ability to discriminate between the smell of water and cycloheximide solution, a strong repellent for rodents. Rats were divided into two groups based on their olfactory discriminative abilities. The olfaction positive (+) group achieved 83% +/- 1% correct responses and the distances of the LOT-transected sites from the middle cerebral artery of this group ranged between 0.8 and 2.4 mm (n = 8). The olfaction negative (-) group achieved 48% +/- 1% correct responses and the distances of the LOT-transected sites from the middle cerebral artery ranged between 2.5 and 4.2 mm (n = 10). From these data, we concluded that the site of the LOT critical for olfactory function is located approximately 2.5 mm from the middle cerebral artery. (c) 2012 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.