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Acetylcholine-induced Ciliary Beat of the Human Nasal Mucosa Is Regulated by the Pannexin-1 Channel and Purinergic P2X Receptor.
Am J Rhinol Allergy. 2018 Jul; 32(4):217-227.AJ

Abstract

Background Airway mucociliary transport is an important function for the clearance of inhaled foreign particulates in the respiratory tract. The present study aimed at investigating the regulatory mechanism of acetylcholine (Ach)-induced ciliary beat of the human nasal mucosa in ex vivo. Methods The inferior turbinate mucosa was collected from patients with chronic hypertrophic rhinitis during endoscopic surgery. The mucosa was cut into thin strips, and ciliary movement was observed under a phase-contrast light microscope with a high-speed digital video camera. The sample was alternatively subjected to scanning electron microscopic observation. Results Cilia on the turbinate epithelium were well preserved at the ultrastructural level. The baseline ciliary beat frequency (CBF) was 6.45 ± 0.32 Hz. CBF was significantly increased by stimulation with 100 µM Ach and 100 µM adenosine triphosphate. The Ach-induced CBF increase was completely inhibited by removing extracellular Ca2+. Significant inhibition of the Ach-induced CBF was also observed by the addition of 1 µM atropine, 40 µM 2-aminoethoxydiphenyl borate (inositol trisphosphate [IP3] receptor antagonist), 10 µM carbenoxolone (pannexin-1 blocker), 1 mM probenecid (pannexin-1 blocker), 100 µM pyridoxalphosphate-6-azophenyl-20,40-disulfonic acid (P2X antagonist), and 300 µM flufenamic acid (connexin blocker). Meanwhile, 30 nM bafilomycin A1 (vesicular transport inhibitor) did not inhibit the Ach-induced CBF increase.

CONCLUSIONS

These results indicate that the regulatory mechanism of the Ach-induced ciliary beat is dependent on extracellular Ca2+ and involves the muscarinic Ach receptor, IP3 receptor, pannexin-1 channel, purinergic P2X receptor, and connexin channel. We proposed a tentative intracellular signaling pathway of the Ach-induced ciliary beat, in which the pannexin-1-P2X unit may play a central role in ciliary beat regulation.

Authors+Show Affiliations

1 Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan. 2 Department of Otorhinolaryngology, Ha Noi Medical University, Ha Noi, Vietnam.1 Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.1 Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.1 Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.3 Shared-Use Research Center, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.1 Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.1 Department of Otorhinolaryngology-Head and Neck Surgery, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29676177

Citation

Do, Ba H., et al. "Acetylcholine-induced Ciliary Beat of the Human Nasal Mucosa Is Regulated By the Pannexin-1 Channel and Purinergic P2X Receptor." American Journal of Rhinology & Allergy, vol. 32, no. 4, 2018, pp. 217-227.
Do BH, Ohbuchi T, Wakasugi T, et al. Acetylcholine-induced Ciliary Beat of the Human Nasal Mucosa Is Regulated by the Pannexin-1 Channel and Purinergic P2X Receptor. Am J Rhinol Allergy. 2018;32(4):217-227.
Do, B. H., Ohbuchi, T., Wakasugi, T., Koizumi, H., Yokoyama, M., Hohchi, N., & Suzuki, H. (2018). Acetylcholine-induced Ciliary Beat of the Human Nasal Mucosa Is Regulated by the Pannexin-1 Channel and Purinergic P2X Receptor. American Journal of Rhinology & Allergy, 32(4), 217-227. https://doi.org/10.1177/1945892418770292
Do BH, et al. Acetylcholine-induced Ciliary Beat of the Human Nasal Mucosa Is Regulated By the Pannexin-1 Channel and Purinergic P2X Receptor. Am J Rhinol Allergy. 2018;32(4):217-227. PubMed PMID: 29676177.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Acetylcholine-induced Ciliary Beat of the Human Nasal Mucosa Is Regulated by the Pannexin-1 Channel and Purinergic P2X Receptor. AU - Do,Ba H, AU - Ohbuchi,Toyoaki, AU - Wakasugi,Tetsuro, AU - Koizumi,Hiroki, AU - Yokoyama,Mitsuru, AU - Hohchi,Nobusuke, AU - Suzuki,Hideaki, Y1 - 2018/04/20/ PY - 2018/4/21/pubmed PY - 2018/11/21/medline PY - 2018/4/21/entrez KW - ATP KW - P2X receptor KW - acetylcholine KW - ciliary beat frequency KW - connexin KW - human nasal mucosa KW - inferior turbinate KW - mucociliary transport KW - pannexin-1 SP - 217 EP - 227 JF - American journal of rhinology & allergy JO - Am J Rhinol Allergy VL - 32 IS - 4 N2 - : Background Airway mucociliary transport is an important function for the clearance of inhaled foreign particulates in the respiratory tract. The present study aimed at investigating the regulatory mechanism of acetylcholine (Ach)-induced ciliary beat of the human nasal mucosa in ex vivo. Methods The inferior turbinate mucosa was collected from patients with chronic hypertrophic rhinitis during endoscopic surgery. The mucosa was cut into thin strips, and ciliary movement was observed under a phase-contrast light microscope with a high-speed digital video camera. The sample was alternatively subjected to scanning electron microscopic observation. Results Cilia on the turbinate epithelium were well preserved at the ultrastructural level. The baseline ciliary beat frequency (CBF) was 6.45 ± 0.32 Hz. CBF was significantly increased by stimulation with 100 µM Ach and 100 µM adenosine triphosphate. The Ach-induced CBF increase was completely inhibited by removing extracellular Ca2+. Significant inhibition of the Ach-induced CBF was also observed by the addition of 1 µM atropine, 40 µM 2-aminoethoxydiphenyl borate (inositol trisphosphate [IP3] receptor antagonist), 10 µM carbenoxolone (pannexin-1 blocker), 1 mM probenecid (pannexin-1 blocker), 100 µM pyridoxalphosphate-6-azophenyl-20,40-disulfonic acid (P2X antagonist), and 300 µM flufenamic acid (connexin blocker). Meanwhile, 30 nM bafilomycin A1 (vesicular transport inhibitor) did not inhibit the Ach-induced CBF increase. CONCLUSIONS: These results indicate that the regulatory mechanism of the Ach-induced ciliary beat is dependent on extracellular Ca2+ and involves the muscarinic Ach receptor, IP3 receptor, pannexin-1 channel, purinergic P2X receptor, and connexin channel. We proposed a tentative intracellular signaling pathway of the Ach-induced ciliary beat, in which the pannexin-1-P2X unit may play a central role in ciliary beat regulation. SN - 1945-8932 UR - https://www.unboundmedicine.com/medline/citation/29676177/Acetylcholine_induced_Ciliary_Beat_of_the_Human_Nasal_Mucosa_Is_Regulated_by_the_Pannexin_1_Channel_and_Purinergic_P2X_Receptor_ L2 - http://journals.sagepub.com/doi/full/10.1177/1945892418770292?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -