Density of α4β2* nAChR on the surface of neurons is modulated by chronic antagonist exposure.
The expression of high-affinity α4β2* nicotinic acetylcholine receptors (nAChR) increases following chronic exposure to nicotinic agonists. While, nAChR antagonists can also produce upregulation, these changes are often less pronounced than achieved with agonists. It is unknown if nAChR agonists and antagonists induce receptor upregulation by the same mechanisms. In this study, primary neuronal cultures prepared from cerebral cortex, hippocampus, diencephalon, and midbrain/hindbrain of C57BL/6J mouse embryos were treated chronically with nicotine (agonist), mecamylamine (noncompetitive antagonist) or dihydro-β-erythroidine (competitive antagonist) or the combination of nicotine with each antagonist. The distribution of intracellular and surface [(125)I]epibatidine-binding sites were subsequently measured. Treatment with 1 μmol/L nicotine upregulated intracellular and cell surface [(125)I]epibatidine binding after 96 h. Chronic dihydro-β-erythroidine (10 μmol/L) treatment also increased [(125)I]epibatidine binding on the cell surface; however, mecamylamine was ineffective in upregulating receptors by itself. The combination of 1 μmol/L nicotine plus 10 μmol/L mecamylamine elicited a significantly higher upregulation than that achieved by treatment with nicotine alone due to an increase of [(125)I]epibatidine binding on the cell surface. This synergistic effect of mecamylamine and nicotine was found in neuronal cultures from all four brain regions. Chronic treatment with nicotine concentrations as low as 10 nmol/L produced upregulation of [(125)I]epibatidine binding. However, the effect of mecamylamine was observed only after coincubation with nicotine concentrations equal to or greater than 100 nmol/L. Vesicular trafficking was required for both nicotine and nicotine plus mecamylamine-induced upregulation. Results presented here support the idea of multiple mechanisms for nAChR upregulation.
Institute for Behavioral Genetics, University of Colorado Boulder, Colorado.
Cristian A
CA
Zambrano
Institute for Behavioral Genetics, University of Colorado Boulder, Colorado.
Caitlin A
CA
Short
Institute for Behavioral Genetics, University of Colorado Boulder, Colorado.
Rakel M
RM
Salamander
Institute for Behavioral Genetics, University of Colorado Boulder, Colorado.
Sharon R
SR
Grady
Institute for Behavioral Genetics, University of Colorado Boulder, Colorado.
Michael J
MJ
Marks
[125I]epibatidine; dihydro-β-erythroidine; mecamylamine; neuronal culture; nicotine; nicotinic receptor