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Rare Human Nicotinic Acetylcholine Receptor α4 Subunit (CHRNA4) Variants Affect Expression and Function of High-Affinity Nicotinic Acetylcholine Receptors
- Source :
- Journal of Pharmacology and Experimental Therapeutics. 348:410-420
- Publication Year :
- 2014
- Publisher :
- American Society for Pharmacology & Experimental Therapeutics (ASPET), 2014.
-
Abstract
- Nicotine, the primary psychoactive component in tobacco smoke, produces its behavioral effects through interactions with neuronal nicotinic acetylcholine receptors (nAChRs). α 4 β 2 nAChRs are the most abundant in mammalian brain, and converging evidence shows that this subtype mediates the rewarding and reinforcing effects of nicotine. A number of rare variants in the CHRNA4 gene that encode the α 4 nAChR subunit have been identified in human subjects and appear to be underrepresented in a cohort of smokers. We compared three of these variants ( α 4R336C, α 4P451L, and α 4R487Q) to the common variant to determine their effects on α 4 β 2 nAChR pharmacology. We examined [ 3 H]epibatidine binding, interacting proteins, and phosphorylation of the α 4 nAChR subunit with liquid chromatography and tandem mass spectrometry (LC-MS/MS) in HEK 293 cells and voltage-clamp electrophysiology in Xenopus laevis oocytes. We observed significant effects of the α 4 variants on nAChR expression, subcellular distribution, and sensitivity to nicotine-induced receptor upregulation. Proteomic analysis of immunopurified α 4 β 2 nAChRs incorporating the rare variants identified considerable differences in the intracellular interactomes due to these single amino acid substitutions. Electrophysiological characterization in X. laevis oocytes revealed alterations in the functional parameters of activation by nAChR agonists conferred by these α 4 rare variants, as well as shifts in receptor function after incubation with nicotine. Taken together, these experiments suggest that genetic variation at CHRNA4 alters the assembly and expression of human α 4 β 2 nAChRs, resulting in receptors that are more sensitive to nicotine exposure than those assembled with the common α 4 variant. The changes in nAChR pharmacology could contribute to differences in responses to smoked nicotine in individuals harboring these rare variants.
- Subjects :
- Cellular and Molecular
Nicotine
Pyridines
Protein subunit
Receptors, Nicotinic
Biology
Xenopus laevis
medicine
Animals
Humans
Nicotinic Agonists
Phosphorylation
Receptor
Acetylcholine receptor
Pharmacology
Polymorphism, Genetic
Cell Membrane
HEK 293 cells
Bridged Bicyclo Compounds, Heterocyclic
Up-Regulation
Cell biology
Protein Subunits
Nicotinic acetylcholine receptor
HEK293 Cells
Nicotinic agonist
Biochemistry
Epibatidine
Oocytes
Molecular Medicine
Female
sense organs
medicine.drug
Subjects
Details
- ISSN :
- 15210103 and 00223565
- Volume :
- 348
- Database :
- OpenAIRE
- Journal :
- Journal of Pharmacology and Experimental Therapeutics
- Accession number :
- edsair.doi.dedup.....df10aa204ae6c98da3e737796998333a