Functional alpha6-containing nicotinic receptors are present in chick retina.
Despite the fact that the neuronal chick alpha6 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 alpha6-containing nicotinic receptors in neuronal tissues. Using subunit-specific antibodies directed against two different epitopes of the chick alpha6 subunit, we performed immunoprecipitation experiments on immunopurified alpha6-containing receptors radiolabeled with the nicotinic agonist [3H]epibatidine (Epi): almost all of the alpha6 receptors contained the beta4 subunit, 51% the beta3 subunit, 42% the alpha3 subunit, and 7.5% the beta2 subunit. Western blot analyses of the purified receptors confirmed the presence of the alpha3, beta3, beta2, and beta4 subunits, and the absence of the alpha4, alpha5, and alpha7 subunits. The alpha6-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 alpha6 receptors also have a distinct antagonist pharmacological profile with a rank order of potency of alpha-conotoxin MII > methyllycaconitine > dihydro-beta-erythroydine > MG624 > d-tubocurarine > decamethonium > hexamethonium. When reconstituted in lipid bilayers, the alpha6-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.
Vailati
S
S
Consiglio Nazionale delle Ricerche (CNR) Cellular and Molecular Pharmacology Center, Department of Medical Pharmacology, University of Milan, Italy.
Hanke
W
W
Bejan
A
A
Barabino
B
B
Longhi
R
R
Balestra
B
B
Moretti
M
M
Clementi
F
F
Gotti
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D002645:Chickens; D008051:Lipid Bilayers / Q000378:metabolism; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D012160:Retina / Q000378:metabolism* / Q000502:physiology; D003336:Tectum Mesencephali / Q000378:metabolism / Q000502:physiology
D000906:Antibodies; D008051:Lipid Bilayers; D011978:Receptors, Nicotinic