title: 5-Iodo-A-85380, a specific ligand for alpha 4 beta 2 nicotinic acetylcholine receptors, prevents glutamate neurotoxicity in rat cortical cultured neurons.
abstract: 5-iodo-3-(2(S)-azetidinylmethoxy)pyridine (5-iodo-A-85380, 5IA) has very high affinity and selectivity to nicotinic acetylcholine receptor (nAChR) alpha 4 beta 2 subtype, and a relative safe profile. To assess whether 5IA has neuroprotective properties, we examined the effect of 5IA on glutamate (Glu)-induced neurotoxicity using primary cultures of rat cortical neurons. A 10-min exposure of cultures to Glu followed by 2-h incubation with drug-free medium caused a marked loss of viability, as determined by trypan blue exclusion method. Glu-induced neurotoxicity was prevented by 5IA both in a time- and concentration-dependent manner. 5IA-induced neuroprotection required pretreatment of 5IA prior to Glu exposure with an optimal concentration of 10 nM and an optimal pretreatment time of 2 h. Treatment after Glu exposure could not rescue the cultured cells. The neuroprotective effect of 5IA was antagonized by mecamylamine, a nAChR antagonist, but not by scopolamine, a muscarinic acetylcholine receptor antagonist. Dihydro-beta-erythroidine, an alpha 4 beta 2 nAChR antagonist, completely inhibited 5IA-induced neuroprotection, whereas alpha-bungarotoxin, an alpha 7 nAChR antagonist, had no effect. Furthermore, 5IA did not show neuroprotective effects in the absence of extracellular Ca2+. These results suggest that the neuroprotective effects of 5IA are produced by activation of alpha 4 beta 2 nAChRs followed by the influx of extracellular Ca2+. In conclusion, 5IA is possibly not only useful for the treatment and prevention of glutamate neurotoxicity, but also as an available tool for elucidating the mechanism of neuroprotection associated with alpha 4 beta 2 nAChRs.
authors.affiliation: Radioisotopes Research Laboratory, Kyoto University Hospital, Faculty of Medicine, Kyoto University, Kyoto 606-8507, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Ueda
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Iida
authors.forename: Youji
authors.initials: Y
authors.lastname: Kitamura
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Kawashima
authors.forename: Mikako
authors.initials: M
authors.lastname: Ogawa
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Magata
authors.forename: Hideo
authors.initials: H
authors.lastname: Saji
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001384:Azetidines / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D018680:Cholinergic Antagonists / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004622:Embryo, Mammalian; D018698:Glutamic Acid / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009498:Neurotoxins / Q000633:toxicity*; D011725:Pyridines / Q000494:pharmacology*; D051381:Rats; D013997:Time Factors
chemical_list: C426745:5-iodo-3-(2-azetidinylmethoxy)pyridine; D001384:Azetidines; D018680:Cholinergic Antagonists; D018696:Neuroprotective Agents; D009498:Neurotoxins; D011725:Pyridines; D018698:Glutamic Acid; D002118:Calcium