title: An increase in intracellular free calcium ions by nicotinic acetylcholine receptors in a single cultured rat cortical astrocyte.
abstract: Neuronal nicotinic acetylcholine receptors (nAChRs) are composed of an assembly between at least seven alpha (alpha2-alpha7, alpha9) and three beta (beta2-beta4) subunits in mammals. The addition of 50 mM KCl or 1 mM nicotine immediately increased the number of cells with high fluorescence intensity in rat cortical astrocytes on fluo-3 fluorescence measurement. Nicotine was effective at increasing the fluorescence intensity in astrocytes cultured for 2 days after replating, but not in those used 1 or 5 days after replating, without markedly affecting the cellular viability irrespective of the exposure period. Nicotine markedly increased the fluorescence intensity in a concentration-dependent manner at a concentration range of 10-100 microM in cultured astrocytes when analyzed on a responsive single cell. In these responsive single cells, the increase by nicotine was significantly prevented by the heteromeric alpha4/beta2 subtype antagonist dihydro-beta-erythroidine and the homomeric alpha7 subtype antagonist methyllycaconitine, as well as by nifedipine and EGTA but not thapsigargin. Methyllycaconitine failed to inhibit further the increase by nicotine in the presence of nifedipine, however, whereas the expression of mRNA was seen for all mammalian neuronal nAChR subunits in cultured rat cortical astrocytes as well as neurons. These results suggest that nicotine may increase intracellular free Ca2+ through the influx of extracellular Ca2+ across L-type voltage-gated Ca2+ channels rather than Ca2+ release from intracellular stores, in a manner related to the alpha4/beta2 and/or alpha7 nAChR channels functionally expressed in cultured rat cortical astrocytes.
authors.affiliation: Laboratory of Molecular Pharmacology, Division of Pharmaceutical Sciences, Kanazawa University Graduate School of Natural Science and Technology, Kanazawa, Ishikawa, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Oikawa
authors.forename: Noritaka
authors.initials: N
authors.lastname: Nakamichi
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kambe
authors.forename: Masato
authors.initials: M
authors.lastname: Ogura
authors.forename: Yukio
authors.initials: Y
authors.lastname: Yoneda
mesh_terms: D000814:Aniline Compounds / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D005110:Extracellular Space / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D009538:Nicotine / Q000494:pharmacology; D018722:Nicotinic Agonists / Q000494:pharmacology; D011189:Potassium Chloride / Q000494:pharmacology; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D011978:Receptors, Nicotinic / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors; D012668:Trigeminal Ganglion / Q000187:drug effects / Q000378:metabolism; D014966:Xanthenes / Q000378:metabolism
chemical_list: D000814:Aniline Compounds; D002121:Calcium Channel Blockers; D018722:Nicotinic Agonists; D021122:Protein Subunits; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D014966:Xanthenes; C059715:Fluo-3; D011189:Potassium Chloride; D009538:Nicotine; D002118:Calcium