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Vol. 56, Issue 1, 11-19, July 1999
6-Containing Nicotinic Receptors Are Present in
Chick Retina
Consiglio Nazionale delle Ricerche (CNR) Cellular and Molecular
Pharmacology Center, Department of Medical Pharmacology, University of
Milan, Milan, Italy (S.V., A.B., Ba.B., M.M., F.C., C.G.); Institute of
Zoophysiology, Hohenheim University, Stuttgart, Germany (W.H.);
Department of Experimental Medicine and Pathology, Università di
Roma "La Sapienza," Rome, Italy (Be.B.); and CNR Center of
Hormone Chemistry, Milan, Italy (R.L.)
Despite the fact that the neuronal chick 
6 subunit was first cloned
several years ago and recently has been shown to form acetylcholine
(ACh)-activated channels in heterologous systems, no information is yet
available concerning the structure and function of the 6-containing
nicotinic receptors in neuronal tissues. Using subunit-specific
antibodies directed against two different epitopes of the chick 6
subunit, we performed immunoprecipitation experiments on immunopurified
6-containing receptors radiolabeled with the nicotinic agonist
[3H]epibatidine (Epi): almost all of the 6 receptors
contained the 
4 subunit, 51% the 3 subunit, 42% the 3
subunit, and 7.5% the 2 subunit. Western blot analyses of the
purified receptors confirmed the presence of the 3, 3, 2, and
4 subunits, and the absence of the 4, 5, and 7 subunits.
The 6-containing receptors bind [3H]Epi
(Kd = 35 pM) and a number of other
nicotinic agonists with very high affinity, the rank order being Epi
cytisine > nicotine > 1,1-dimethyl-4-phenylpiperazinium > acetylcholine > carbamylcholine. The 6 receptors also have a distinct antagonist
pharmacological profile with a rank order of potency of -conotoxin
MII > methyllycaconitine > dihydro--erythroydine > MG624 > d-tubocurarine > decamethonium > hexamethonium. When reconstituted in lipid bilayers, the
6-containing receptors form functional cationic channels with a main
conductance state of 48 pS. These channels are activated by nicotinic
agonists in a dose-dependent manner, and blocked by the nicotinic
antagonist d-tubocurarine.
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