title: Chronic nicotine treatment up-regulates alpha3 and alpha7 acetylcholine receptor subtypes expressed by the human neuroblastoma cell line SH-SY5Y.
abstract: Chronic exposure to nicotine has been reported to increase the number of nicotinic acetylcholine receptors (AChRs) in brain. The mechanism of up-regulation for the alpha4beta2 AChR subtype, which accounts for the majority of high affinity nicotine binding in mammalian brain, has previously been shown to involve a decrease in the rate of alpha4beta2 AChR turnover. Here, we report an investigation of the extent and mechanism of nicotine-induced up-regulation of alpha3 AChRs and alpha7 AChR subtypes expressed in the human neuroblastoma cell line SH-SY5Y. Up-regulation of human alpha3 AChRs and alpha7 AChRs, unlike alpha4beta2 AChRs, requires much higher nicotine concentrations than are encountered in smokers; the extent of increase of surface AChRs is much less; and the mechanisms of up-regulation are different than with alpha4beta2 AChRs. The mechanisms of up-regulation may be different for alpha3 AChRs or alpha7 AChRs. Chronic treatment with nicotine or carbamylcholine, but not d-tubocurarine, mecamylamine, or dihydro-beta-erythroidine, induced a 500-600% increase in the number of alpha3 AChRs but only a 30% increase in alpha7 AChRs. Chronic nicotine treatment did not increase affinity for nicotine or increase the amount of RNA for alpha3 or alpha7 subunits. The effect of nicotine on up-regulation of alpha7 AChRs was partially blocked by either d-tubocurarine or mecamylamine. The effect of nicotine treatment on the number of alpha3 AChRs was only slightly blocked by the antagonists d-tubocurarine, mecamylamine, or dihydro-beta-erythroidine at concentrations that efficiently block alpha3 AChR function. Most of the nicotine-induced increase in alpha3 AChRs was found to be intracellular. The alpha3 AChRs, which accumulate intracellularly, were shown to have been previously exposed on the cell surface by their susceptibility to antigenic modulation. The data suggest that chronic exposure to nicotine may induce a conformation of cell surface alpha3 AChRs that at least in this cell line are consequently internalized but not immediately destroyed.
authors.affiliation: Department of Neuroscience, University of Pennsylvania Medical School, Philadelphia 19104-6074, USA.
authors.forename: X
authors.initials: X
authors.lastname: Peng
authors.forename: V
authors.initials: V
authors.lastname: Gerzanich
authors.forename: R
authors.initials: R
authors.lastname: Anand
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.forename: J
authors.initials: J
authors.lastname: Lindstrom
mesh_terms: D004705:Endocytosis; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology; D009538:Nicotine / Q000008:administration & dosage / Q000494:pharmacology*; D011950:Receptors, Cholinergic / Q000145:classification / Q000187:drug effects* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000187:drug effects*
chemical_list: D011950:Receptors, Cholinergic; D009538:Nicotine