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Vol. 54, Issue 6, 1124-1131, December 1998
4
2 Nicotinic
Acetylcholine Receptor Expressed in Xenopus laevis
Oocytes
Research Institute of Toxicology, Utrecht University, Utrecht, The
Netherlands
Nicotinic receptors generally are presumed to consist of two 
and
three non- subunits. We varied the relative levels of expression of
the neuronal nicotinic 4 and 
2 receptor subunits in
Xenopus laevis oocytes by nuclear injection of cDNAs
coding for these subunits in : ratios of 9:1, 1:1, and 1:9. The
sensitivities of the receptors to acetylcholine and
d-tubocurarine were investigated in voltage-clamp
experiments. For receptors expressed at the 9:1 and 1:1 : ratios,
the EC50 value of acetylcholine is ~60 µM. For the majority of the receptors expressed at the 1:9 : ratio, the sensitivity to acetylcholine is enhanced 30-fold. No evidence for
more than one type of acetylcholine binding site in a single receptor
is obtained. The sensitivity to d-tubocurarine decreases with decreasing : ratio. IC50 values of
d-tubocurarine are 0.2, 0.5, and 2 µM for
the 9:1, 1:1, and 1:9 : ratios, respectively. At the 1:9 :
ratio, additional receptors with an IC50 value of 163 µM d-tubocurarine are expressed. At least
two components with distinct sensitivities to
d-tubocurarine are required to account for the shift in
IC50. The combined agonist and antagonist effects reveal
four distinct subtypes of 42 nicotinic receptors. The results
imply that the subunit stoichiometry of heteromeric 42
acetylcholine receptors is not restricted to 2:3.
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