Mammalian nicotinic receptors with alpha7 subunits that slowly desensitize and rapidly recover from alpha-bungarotoxin blockade.
One of the most abundant nicotinic receptors in the nervous system is a species that contains the alpha7 gene product, rapidly desensitizes, and binds alpha-bungarotoxin with great affinity. The receptor has a high relative permeability to calcium and performs a variety of functions including presynaptic modulation of transmitter release and postsynaptic generation of synaptic currents. Fast excitatory transmission in mammalian intracardiac ganglia is mediated primarily by nicotinic receptors, and although intracardiac ganglion neurons express the alpha7 gene, no toxin-sensitive response has been detected previously in them. We report here that whole-cell patch-clamp recordings from freshly dissociated intracardiac ganglion neurons reveal a nicotinic response that desensitizes slowly and is blocked by alpha-bungarotoxin in a rapidly reversible manner. The only rat gene previously thought capable of forming such receptors was alpha9, but no evidence suggests that the alpha9 gene is expressed in neurons. We find that reverse transcription (RT)-PCR detects alpha7 but not alpha9 mRNA in the ganglia. In addition, the pharmacology of the nicotinic response is typical of alpha7-containing receptors but differs in several respects from that expected for alpha9. Binding experiments with immunotethered receptors identifies a ganglionic species that contains the alpha7 gene product. Moreover, intracellular perfusion of the cells with an anti-alpha7 monoclonal antibody specifically reduces the amplitude of the toxin-sensitive response. The results indicate that alpha7-containing receptors are responsible for the slowly desensitizing, toxin-reversible response and suggest that the receptors are modified in cell-specific ways to influence their functional properties.
Cuevas
J
J
Department of Biology, University of California, San Diego, La Jolla, California 92093-0357, USA.
Berg
D K
DK
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D000831:Animals, Newborn; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002038:Bungarotoxins / Q000494:pharmacology*; D002478:Cells, Cultured; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D005071:Evoked Potentials / Q000187:drug effects / Q000502:physiology; D005726:Ganglia, Parasympathetic / Q000502:physiology; D006321:Heart / Q000294:innervation; D066298:In Vitro Techniques; D009474:Neurons / Q000187:drug effects / Q000502:physiology; D018733:Nicotinic Antagonists / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011978:Receptors, Nicotinic / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D064569:alpha7 Nicotinic Acetylcholine Receptor
D000911:Antibodies, Monoclonal; D002038:Bungarotoxins; C579165:Chrna7 protein, rat; D018733:Nicotinic Antagonists; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; D000109:Acetylcholine