Exposure to nicotine alters the trafficking and assembly of nicotinic receptors (nAChRs), leading to their up-regulation on the plasma membrane. Although the mechanism is not fully understood, nicotine-induced up-regulation is believed to contribute to nicotine addiction. The effect of cotinine, the primary metabolite of nicotine, on nAChR trafficking and assembly has not been extensively investigated. We utilize a pH-sensitive variant of GFP, super ecliptic pHluorin, to differentiate between intracellular nAChRs and those expressed on the plasma membrane to quantify changes resulting from cotinine and nicotine exposure. Similar to nicotine, exposure to cotinine increases the number of α4β2 receptors on the plasma membrane and causes a redistribution of intracellular receptors. In contrast to this, cotinine exposure down-regulates α6β2β3 receptors. We also used single molecule fluorescence studies to show that cotinine and nicotine both alter the assembly of α4β2 receptors to favor the high sensitivity (α4)2(β2)3 stoichiometry.

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Published in The Journal of Biological Chemistry, v. 290, no. 40, p. 24403-24412.

This research was originally published in The Journal of Biological Chemistry. Ashley M. Fox, Faruk H. Moonschi, and Christopher I. Richards. The Nicotine Metabolite, Cotinine, Alters the Assembly and Trafficking of a Subset of Nicotinic Acetylcholine Receptors. The Journal of Biological Chemistry. 2015; 290:24403-24412. © the American Society for Biochemistry and Molecular Biology.

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This work was supported in part by the National Institute on Drug Abuse (NIDA) Training Grant DA016176 (to A. M. F.) and NIDA Grant DA038817 (C. I. R.). The authors declare that they have no conflicts of interest with the contents of this article.

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