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Next Article 
Volume 17, Number 5,
Issue of March 1, 1997
pp. 1531-1538
Copyright ©1997 Society for Neuroscience
Formation of Oligomers Containing the  3 and 4 Subunits of
the Rat Nicotinic Receptor
Received Oct. 7, 1996; revised Nov. 18, 1996; accepted Nov. 26, 1996.
John R. Forsayeth and
Eugenia Kobrin
Department of Anesthesia, University of California San Francisco,
San Francisco, California 94143-0542
The role of the  3 and 4 subunits of the nicotinic
acetylcholine receptor in brain is still unclear. We investigated
nicotinic receptor structure with antibodies directed against unique
regions of the 3 and 4 subunits of the rat nicotinic
acetylcholine receptor. Anti-4 detected a single band of 66 kDa in
most regions of the brain that was strongest in striatum and
cerebellum. The 60 kDa 3 subunit was detected primarily in striatum
and cerebellum, and faintly in hippocampus. Immunoprecipitation
experiments established that the two subunits were coassembled in the
cerebellum along with the 2 subunit. Antibodies against the  4,
2, 3, and 4 subunits immunoprecipitated ~75% of the
bungarotoxin-insensitive nicotinic receptor from cerebellar extracts as
determined by nicotine-dependent acetylcholine binding. Transfection of
COS cells with cDNAs for these four subunits induced expression of a
high affinity nicotinic receptor. Omission of only a single subunit
from the transfection affected either the
Bmax or the apparent
KD of the receptor. Our data suggest that
the 3 subunit functions as a structural entity that links a
relatively unstable 42 heterodimer to a more stable 44
heterodimer. The agonist-binding site formed by 42 has a much
greater affinity than does that formed by 44. In this respect,
nicotinic receptors that contain the 3 subunit are structurally homologous to the muscle nicotinic receptor.
Key words:
nicotine;
acetylcholine;
receptor;
antibody;
3;
4;
subunit
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