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Carbocisteine stimulated an increase in ciliary bend angle via a decrease in [Cl]i in mouse airway cilia

  • Yukiko Ikeuchi
  • Haruka Kogiso
  • Shigekuni Hosogi
  • Saori Tanaka
  • Chikao Shimamoto
  • Hitoshi Matsumura
  • Toshio Inui
  • Yoshinori Marunaka
  • Takashi Nakahari
Signaling and cell physiology
Part of the following topical collections:
  1. Signaling and cell physiology

Abstract

Carbocisteine (CCis), a mucoactive agent, is widely used to improve respiratory diseases. This study demonstrated that CCis increases ciliary bend angle (CBA) by 30% and ciliary beat frequency (CBF) by 10% in mouse airway ciliary cells. These increases were induced by an elevation in intracellular pH (pHi; the pHi pathway) and a decrease in the intracellular Cl concentration ([Cl]i; the Cl pathway) stimulated by CCis. The Cl pathway, which is independent of CO2/HCO3, increased CBA by 20%. This pathway activated Cl release via activation of Cl channels, leading to a decrease in [Cl]i, and was inhibited by Cl channel blockers (5-nitro-2-(3-phenylpropylamino) benzoic acid and CFTR(inh)-172). Under the CO2/HCO3-free condition, the CBA increase stimulated by CCis was mimicked by the Cl-free NO3 solution. The pHi pathway, which depends on CO2/HCO3, increased CBF and CBA by 10%. This pathway activated HCO3 entry via Na+/HCO3 cotransport (NBC), leading to a pHi elevation, and was inhibited by 4,4′-diisothiocyano-2,2′-stilbenedisulfonic acid. The effects of CCis were not affected by a protein kinase A inhibitor (1 μM PKI-A) or Ca2+-free solution. Thus, CCis decreased [Cl]i via activation of Cl channels including CFTR, increasing CBA by 20%, and elevated pHi via NBC activation, increasing CBF and CBA by 10%.

Keywords

Airway cilia Intracellular Cl concentration Ciliary beating angle Inner dynein 

Notes

Acknowledgements

We thank Osaka Medical College for giving us an opportunity to perform the experiments using the video microscope equipped with a high-speed camera.

Funding

This work was partly supported by Grants-in-Aid for Scientific Research from the Japan Society of the Promotion of Science to YM (No. JP18H03182) and to SH (No. 17K08545).

Compliance with ethical standards

The procedures and protocols for the experiments were approved by the Committee for Animal Research of Kyoto Prefectural University of Medicine (No. 26-263) and Ritsumeikan University (No. BKC 2017-050). The animals were cared for, and the experiments were carried out according to the guidelines of this committee. Female mice (C57BL/6J, 6 weeks of age) were purchased from Shimizu Experimental Animals (Kyoto, Japan) and fed standard pellet food and water ad libitum.

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Yukiko Ikeuchi
    • 1
    • 2
  • Haruka Kogiso
    • 1
    • 2
  • Shigekuni Hosogi
    • 1
  • Saori Tanaka
    • 3
  • Chikao Shimamoto
    • 3
  • Hitoshi Matsumura
    • 3
  • Toshio Inui
    • 2
    • 4
  • Yoshinori Marunaka
    • 1
    • 2
    • 5
  • Takashi Nakahari
    • 2
  1. 1.Department of Molecular Cell Physiology, Graduate School of Medical ScienceKyoto Prefectural University of MedicineKyotoJapan
  2. 2.Research Center for Drug Discovery and Pharmaceutical Development Science, Research Organization of Science and Technology, BKCRitsumeikan UniversityKusatsuJapan
  3. 3.Laboratory of PharmacotherapyOsaka University of Pharmaceutical SciencesTakatsukiJapan
  4. 4.Saisei Mirai ClinicsMoriguchiJapan
  5. 5.Research Institute for Clinical PhysiologyKyoto Industrial Health AssociationKyotoJapan

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