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

  • Signaling and cell physiology
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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%.

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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).

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Authors and Affiliations

  1. Department of Molecular Cell Physiology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, 602-8566, Japan

    Yukiko Ikeuchi, Haruka Kogiso, Shigekuni Hosogi & Yoshinori Marunaka

  2. Research Center for Drug Discovery and Pharmaceutical Development Science, Research Organization of Science and Technology, BKC, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga, 525-8577, Japan

    Yukiko Ikeuchi, Haruka Kogiso, Toshio Inui, Yoshinori Marunaka & Takashi Nakahari

  3. Laboratory of Pharmacotherapy, Osaka University of Pharmaceutical Sciences, Takatsuki, Japan

    Saori Tanaka, Chikao Shimamoto & Hitoshi Matsumura

  4. Saisei Mirai Clinics, Moriguchi, 570-0012, Japan

    Toshio Inui

  5. Research Institute for Clinical Physiology, Kyoto Industrial Health Association, Kyoto, 604-8472, Japan

    Yoshinori Marunaka

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  1. Yukiko Ikeuchi
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  2. Haruka Kogiso
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  3. Shigekuni Hosogi
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  4. Saori Tanaka
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  6. Hitoshi Matsumura
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  7. Toshio Inui
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  8. Yoshinori Marunaka
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  9. Takashi Nakahari
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Corresponding author

Correspondence to Takashi Nakahari.

Ethics declarations

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.

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The authors declare that they have no conflict of interest.

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Ikeuchi, Y., Kogiso, H., Hosogi, S. et al. Carbocisteine stimulated an increase in ciliary bend angle via a decrease in [Cl−]i in mouse airway cilia. Pflugers Arch - Eur J Physiol 471, 365–380 (2019). https://doi.org/10.1007/s00424-018-2212-2

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