Chick optic lobe contains a developmentally regulated alpha2alpha5beta2 nicotinic receptor subtype.
The most widely expressed neuronal nicotinic acetylcholine receptor subtype in chick brain is that containing the alpha4 and beta2 subunits. However, immunoprecipitation and localization studies have shown that some brain areas also contain the alpha2 and/or alpha5 subunits, whose role in the definition of receptor properties is still intriguing. Using subunit-specific polyclonal antibodies, we found that the optic lobe is the chick central nervous system region that expresses the highest level of alpha2-containing receptors. Immunoprecipitation studies of these immunopurified alpha2-containing receptors labeled with the nicotinic agonist [(3)H]epibatidine showed that almost all of them contained the beta2 subunit and that more than 66% contained the alpha5 subunit. Western blot analyses of the purified receptors confirmed the presence of the alpha2, alpha5, and beta2 subunits and the absence of the alpha3, alpha4, alpha6, alpha7, alpha8, beta3, and beta4 subunits. The alpha2-containing receptors are developmentally regulated: their expression increases 25 times from embryonic day 7 to posthatching day 1 in the optic lobe, compared with an increase of only 5-fold in the forebrain. The alpha2-containing optic lobe receptors bind [(3)H]epibatidine (K(d) = 29 pM) and a number of other nicotinic agonists with very high affinity and have a pharmacological profile very similar to that of the alpha4beta2 subtype. They form functional cationic channels when reconstituted in lipid bilayers, with pharmacological and biophysical properties different from those of the alpha4beta2 subtype. These channels are activated by nicotinic agonists in a dose-dependent manner and are blocked by the nicotinic antagonist d-tubocurarine.
Balestra
B
B
Consiglio Nazionale delle Ricerche Cellular and Molecular Pharmacology Center, Department of Medical Pharmacology, University of Milan, Milan, Italy.
Vailati
S
S
Moretti
M
M
Hanke
W
W
Clementi
F
F
Gotti
C
C
D000818:Animals; D000906:Antibodies / Q000276:immunology; D015153:Blotting, Western; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002490:Central Nervous System / Q000187:drug effects / Q000378:metabolism; D002645:Chickens; D018507:Gene Expression Regulation, Developmental; D066298:In Vitro Techniques; D008051:Lipid Bilayers / Q000378:metabolism; D018722:Nicotinic Agonists / Q000494:pharmacology; D011233:Precipitin Tests; D016548:Prosencephalon / Q000187:drug effects / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D011978:Receptors, Nicotinic / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D003336:Tectum Mesencephali / Q000378:metabolism*; D014316:Tritium
D000906:Antibodies; D019086:Bridged Bicyclo Compounds, Heterocyclic; D008051:Lipid Bilayers; D018722:Nicotinic Agonists; D011725:Pyridines; D011978:Receptors, Nicotinic; C403531:nicotinic receptor alpha4beta2; D014316:Tritium; C082748:epibatidine