Epithelium-dependent inhibition of cholinergic transmission in rat isolated trachea by potassium channel openers.
We have investigated the effects of the potassium channel openers, cromakalim and pinacidil, on cholinergic transmission in rat airways. Experiments were performed on epithelium-intact and epithelium-denuded preparations of rat isolated trachea which had been incubated with [3H]-choline to incorporate [3H]-acetylcholine into the cholinergic transmitter stores. In radiolabelled, epithelium-intact preparations, electrical field stimulation (60 s trains of 1 ms pulses, 5 Hz, 15 V) evoked an efflux of radioactivity that was unaffected by the removal of extracellular Ca2+ and, a large proportion of which was resistant to tetrodoxin (1 microM). In contrast, in epithelium-denuded preparations, both tetrodotoxin and Ca2+ withdrawal virtually abolished the stimulation-induced (S-I) efflux. Thus, with epithelium-denuded but not with epithelium-intact tracheal preparations, the S-1 efflux reflects the release of [3H]-acetylcholine from cholinergic nerves. Atropine (1 microM) markedly enhanced the S-I efflux in both epithelium-intact and epithelium-denuded preparations. In epithelium-intact preparations, the combination of atropine (1 microM) and tetrodotoxin (1 microM) reduced the S-I efflux to about the same level as did tetrodotoxin alone. Thus, in epithelium-intact tracheal preparations, when prejunctional muscarinic cholinoceptors subserving autoinhibition of transmitter release are blocked, S-I efflux may be taken as an index of transmitter acetylcholine release. Cromakalim (1 microM) had no effect on the S-I efflux from either epithelium-intact or epithelium-denuded tracheal preparations. However, in epithelium-intact preparations, when atropine (1 microM) was present, cromakalim (1 and 10 microM) and pinacidil (100 microM) significantly inhibited the S-I efflux. In epithelium-denuded preparations, in the presence of atropine (1 microM), cromakalim (1 microM) and pinacidil (100 microM) were without effect on S-I efflux. The inhibition of S-I efflux produced by cromakalim (1 microM) and pinacidil (100 microM) in epithelium-intact tracheal preparations (in the presence of atropine) was prevented by the ATP-sensitive potassium channel blocking drug glibenclamide (1 microM). Glibenclamide (1 microM) alone enhanced S-I efflux from epithelium-intact preparations in the absence but not in the presence of atropine (1 microM). Glibenclamide (1 microM) was without effect on S-I efflux in epithelium-denuded preparations both in the absence or presence of atropine (1 microM). In conclusion, the present study has provided additional evidence of an inhibitory action of the potassium channel openers, cromakalim and pinacidil, on the release of acetylcholine from parasympathetic nerves of the rat trachea which is dependent upon the functional integrity of the airway epithelium. The findings suggest that cromakalim and pinacidil may inhibit transmitter acetylcholine release by opening ATP-sensitive potassium channels, presumably on epithelial cells. In addition, the enhancement of S-I efflux from epithelium-intact tracheal preparations by glibenclamide may indicate that ATP-sensitive potassium channels on epithelial cells play a functional role in the modulation of transmitter acetylcholine release from parasympathetic cholinergic nerves of the airways.
Fabiani
M E
ME
Department of Medical Laboratory Science, Faculty of Biomedical and Health Sciences, RMIT University, Melbourne, Australia.
Vlahos
R
R
Story
D F
DF
D000109:Acetylcholine / Q000378:metabolism*; D000818:Animals; D001285:Atropine / Q000494:pharmacology; D001578:Benzopyrans / Q000494:pharmacology; D019806:Cromakalim; D004848:Epithelium / Q000502:physiology; D005260:Female; D005905:Glyburide / Q000494:pharmacology; D006146:Guanidines / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D020110:Pinacidil; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*; D011758:Pyrroles / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014132:Trachea / Q000187:drug effects* / Q000378:metabolism
D001578:Benzopyrans; D006146:Guanidines; D015221:Potassium Channels; D011758:Pyrroles; D019806:Cromakalim; D020110:Pinacidil; D001285:Atropine; D000109:Acetylcholine; D005905:Glyburide