Acetylcholine-induced ex vivo ATP release from the human nasal mucosa.
OBJECTIVE
The present study aimed at investigating ATP release in response to acetylcholine (Ach) and pharmacologically elucidating the intracellular signal transduction pathway of this reaction in an ex vivo experiment.


METHODS
The inferior turbinate mucosa was collected from 21 patients with chronic hypertrophic rhinitis who underwent endoscopic turbinectomy. The mucosa was shaped into a filmy round piece, and incubated with chemical(s) in Hank's balanced salt solution for 10min. After incubation, the ATP concentration was measured by a luciferin-luciferase assay.


RESULTS
The baseline release of ATP without stimulus was 57.2±10.3fM. The ATP release was significantly increased by stimulation with 100μM Ach. The Ach-induced ATP release was completely inhibited by removing extracellular Ca2+. Significant inhibition of the Ach-induced ATP release was also observed by the addition of 1μM atropine, 40μM 2-APB, 10μM CBX, and 100μM PPADS, whereas 30nM bafilomycin A1 did not affect the ATP release.


CONCLUSIONS
These results indicate that the Ach-induced ATP release from the human nasal mucosa is dependent on the pannexin-1 channel and purinergic P2X7 receptor, suggesting that these two molecules constitute a local autocrine/paracrine signaling system in the human nasal epithelium.
Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Japan.
Hiroki
H
Koizumi
Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Japan.
Shoji
S
Ikezaki
Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Japan.
Toyoaki
T
Ohbuchi
Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Japan.
Ba Hung
BH
Do
Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Japan.
Nobusuke
N
Hohchi
Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Japan.
Rintaro
R
Kawaguchi
Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Japan.
Takuro
T
Kitamura
Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Japan. Electronic address: suzuhyde@med.uoeh-u.ac.jp.
Hideaki
H
Suzuki
D000109:Acetylcholine / Q000494:pharmacology*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000293:Adolescent; D000328:Adult; D000368:Aged; D000897:Anti-Ulcer Agents / Q000494:pharmacology; D001285:Atropine / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002229:Carbenoxolone / Q000494:pharmacology; D018679:Cholinergic Agonists / Q000494:pharmacology*; D017630:Connexins / Q000037:antagonists & inhibitors; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006801:Humans; D018942:Macrolides / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D018727:Muscarinic Antagonists / Q000494:pharmacology*; D009297:Nasal Mucosa / Q000187:drug effects* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors; D058919:Purinergic P2 Receptor Antagonists / Q000494:pharmacology; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D012220:Rhinitis / Q000601:surgery; D015398:Signal Transduction; D055815:Young Adult
D000897:Anti-Ulcer Agents; D018679:Cholinergic Agonists; D017630:Connexins; D004791:Enzyme Inhibitors; D018942:Macrolides; D018727:Muscarinic Antagonists; D009419:Nerve Tissue Proteins; C492764:PANX1 protein, human; D058919:Purinergic P2 Receptor Antagonists; C077792:pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid; D011732:Pyridoxal Phosphate; D001285:Atropine; C040929:bafilomycin A1; D000255:Adenosine Triphosphate; D002229:Carbenoxolone; D000109:Acetylcholine; D002118:Calcium
ATP; Acetylcholine; Human nasal mucosa; Inferior turbinate; P2X7 receptor; Pannexin-1