Pharmacological properties of immuno-isolated neuronal nicotinic receptors.
Recently we immunoaffinity-purified an ACh receptor from chicken brain using a monoclonal antibody raised against receptors from fish electric organ (Whiting and Lindstrom, 1986). This neuronal receptor could be affinity-labeled with 3H-bromoacetylcholine, and antisera to it specifically blocked ACh-induced depolarization of chicken ciliary ganglion cells. Here we show that this neuronal ACh receptor binds 3H-nicotine with high affinity (KD = 6.61 +/- 0.13 nM). 3H-Nicotine binding was blocked by various nicotinic cholinergic ligands but not by alpha-bungarotoxin or the muscarinic antagonist atropine. Binding was also blocked by affinity labeling the receptor with bromoacetylcholine (after reduction by dithiothreitol). Additionally, we were able to use rat antisera raised against the chicken brain receptor to isolate a component from detergent extracts of rat brain that also bound 3H-nicotine with high affinity (KD = 1.5 nM). The pharmacology of this putative ACh receptor from rat brain was almost identical to the receptor from chicken brain, and its regional distribution was in good agreement with that of 3H-nicotine binding to rodent brain membranes reported by other workers. Thus, by analogy to the receptor we have purified and characterized from chicken brain, this nicotine-binding component from rat brain is probably a functional mammalian neuronal nicotinic ACh receptor.
D000818:Animals; D001921:Brain / Q000378:metabolism; D002645:Chickens; D003902:Detergents; D007106:Immune Sera / Q000276:immunology; D007158:Immunologic Techniques; D008024:Ligands / Q000378:metabolism; D009474:Neurons / Q000276:immunology / Q000378:metabolism*; D009538:Nicotine / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D011978:Receptors, Nicotinic / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D014018:Tissue Distribution; D014020:Tissue Extracts
D003902:Detergents; D007106:Immune Sera; D008024:Ligands; D011978:Receptors, Nicotinic; D014020:Tissue Extracts; D009538:Nicotine