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3 and 7
Acetylcholine Receptor Subtypes Expressed by the Human Neuroblastoma
Cell Line SH-SY5Y
Departments of
Neuroscience (X.P., V.G., F.W., J.L.) and
Pharmacology (R.A.), University of Pennsylvania Medical School,
Philadelphia, Pennsylvania 19104-6074
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 
4
2 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 42
AChR turnover. Here, we report an investigation of the extent and
mechanism of nicotine-induced up-regulation of 3 AChRs and 7 AChR
subtypes expressed in the human neuroblastoma cell line SH-SY5Y.
Up-regulation of human 3 AChRs and 7 AChRs, unlike 42
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
42 AChRs. The mechanisms of up-regulation may be different for
3 AChRs or 7 AChRs. Chronic treatment with nicotine or
carbamylcholine, but not d-tubocurarine,
mecamylamine, or dihydro--erythroidine, induced a
500-600% increase in the number of 3 AChRs but only a 30%
increase in 7 AChRs. Chronic nicotine treatment did not increase
affinity for nicotine or increase the amount of RNA for 3 or 7
subunits. The effect of nicotine on up-regulation of 7 AChRs was
partially blocked by either d-tubocurarine or
mecamylamine. The effect of nicotine treatment on the number of 3
AChRs was only slightly blocked by the antagonists
d-tubocurarine, mecamylamine, or
dihydro--erythroidine at concentrations that efficiently block 3
AChR function. Most of the nicotine-induced increase in 3 AChRs was
found to be intracellular. The 3 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 3 AChRs that at least in this cell line are
consequently internalized but not immediately destroyed.
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