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Vol. 60, Issue 6, 1201-1209, December 2001
7
Nicotinic Acetylcholine Receptors
Department of Biochemistry, Molecular Biology, and Biophysics,
University of Minnesota, Minneapolis-St. Paul, Minnesota and Department
of Pharmacology, University of Minnesota, Minneapolis, Minnesota (Y.W.,
A.D.J.M., N.S.O., D.N., S.L., B.M.C.-F.); Department of Pharmacology
and Experimental Therapeutics, University of Maryland School of
Medicine, Baltimore, Maryland (E.F.R.P., E.X.A.); and Institute of
Biophysics Carlos Chagas Filho and Department of Basic and Clinical
Pharmacology, Federal University of Rio de Janeiro, Rio de Janeiro,
Brazil (E.X.A.)
The epithelial or endothelial cells that line the human bronchi and the
aorta express nicotinic acetylcholine receptors (nAChRs) of 
3
subtypes. We report here that human bronchial epithelial cells (BEC)
and aortic endothelial cells (AEC) express also the nAChR 7 subunit,
which forms functional nAChRs. Polymerase chain reaction and in situ
hybridization experiments detected 7 subunit mRNA in cultured human
BEC and AEC and in sections of rat trachea. The binding of radiolabeled
-bungarotoxin revealed a few thousand binding sites per cell in
cultured human BEC and human and bovine AEC. Western blot and
immunohistochemistry experiments demonstrated that cultured BEC and AEC
express a protein(s) recognized by anti-7 antibodies. Whole-cell
patch-clamp studies of cultured human BEC demonstrated the presence of
fast-desensitizing currents activated by choline and nicotine that were
blocked reversibly by methyllycaconitine (1 nM) and irreversibly by
-bungarotoxin (100 nM), consistent with the expression of functional
7 nAChRs. In some cells, choline activated also slowly decaying
currents, confirming previous reports that BEC express functional
3
4 nAChRs. Exposure of cultured BEC to nicotine (1 µM) for 3 days up-regulated functional 7 and 3 nAChRs, as indicated by the
increased number of cells responding to acetylcholine and choline, with
both fast-desensitizing currents, which were blocked irreversibly by
-bungarotoxin, and with slowly desensitizing currents, which are
-bungarotoxin-insensitive currents. The presence of 7
nAChRs in BEC and AEC suggests that some toxic effects of tobacco smoke
could be mediated through these nicotine-sensitive receptors.
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