title: An alpha3beta4 subunit combination acts as a major functional nicotinic acetylcholine receptor in male rat pelvic ganglion neurons.
abstract: We identified major subunits of the nicotinic acetylcholine receptor (nAChR) involved in excitatory postsynaptic potential and intracellular Ca(2+) ([Ca(2+)]i) increase in the major pelvic ganglion (MPG) neurons of the male rat. ACh elicited fast inward currents in both sympathetic and parasympathetic MPG neurons. Mecamylamine, a selective antagonist for alpha3beta4 nAChR, potently inhibited the ACh-induced currents in sympathetic and parasympathetic neurons (IC(50); 0.53 and 0.22 microM, respectively). Furthermore, alpha-conotoxin AuIB (10 microM), a new selective antagonist for alpha3beta4 nAChR, blocked more than 80% of the ACh-induced currents in MPG neurons. Conversely, alpha-bungarotoxin, alpha-methyllycaconitine, and dihydro-beta-erythroidine, known as blockers of the alpha7 or alpha4beta2, did not show selective blocking effects on MPG neurons. ACh transiently increased [Ca(2+)]i which was subsequently abolished in the extracellular Ca(2+)-free environment. Simultaneous recording of [Ca(2+)]i and ionic currents revealed that ACh increased [Ca(2+)]i under the conditions of the voltage-clamped (at -80 mV) state, and this resulted from the influx through nAChR itself. ACh-induced [Ca(2+)]i increase was blocked by mecamylamine (10 microM), but was not affected by atropine (1 microM). RT-PCR analysis showed that, among subunits of nAChR, alpha3 and beta4 were predominantly expressed in MPG. We suggest that activation of alpha3 and beta4 nAChR subunits in MPG neurons induce fast inward currents and [Ca(2+)]i increase, possibly mediating a major role in pelvic autonomic synaptic transmission.
authors.affiliation: Department of Physiology and Institute of Basic Medical Science, Yonsei University Wonju College of Medicine, Wonju, South Korea.
authors.forename: Kyu-Sang
authors.initials: KS
authors.lastname: Park
authors.forename: Seung-Kyu
authors.initials: SK
authors.lastname: Cha
authors.forename: Min-Jeong
authors.initials: MJ
authors.lastname: Kim
authors.forename: Dae-Ran
authors.initials: DR
authors.lastname: Kim
authors.forename: Seong-Woo
authors.initials: SW
authors.lastname: Jeong
authors.forename: Joong-Woo
authors.initials: JW
authors.lastname: Lee
authors.forename: In Deok
authors.initials: ID
authors.lastname: Kong
mesh_terms: D000109:Acetylcholine / Q000037:antagonists & inhibitors / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000502:physiology; D002121:Calcium Channel Blockers; D020916:Conotoxins / Q000494:pharmacology; D004594:Electrophysiology; D005726:Ganglia, Parasympathetic / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005728:Ganglia, Sympathetic / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D066298:In Vitro Techniques; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018733:Nicotinic Antagonists / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D051381:Rats; D011978:Receptors, Nicotinic / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013050:Spectrometry, Fluorescence; D009435:Synaptic Transmission / Q000502:physiology
chemical_list: D002121:Calcium Channel Blockers; D020916:Conotoxins; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic; C403530:nicotinic receptor alpha3beta4; D000109:Acetylcholine; D002118:Calcium