Ciliary beating amplitude controlled by intracellular Cl− and a high rate of CO2 production in ciliated human nasal epithelial cells
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The ciliary transport is controlled by two parameters of the ciliary beating, frequency (CBF) and amplitude. In this study, we developed a novel method to measure both CBF and ciliary bend distance (CBD, an index of ciliary beating amplitude) in ciliated human nasal epithelial cells (cHNECs) in primary culture, which are prepared from patients contracting allergic rhinitis and chronic sinusitis. An application of Cl−-free NO3− solution or bumetanide (an inhibitor of Na+/K+/2Cl− cotransport), which decreases intracellular Cl− concentration ([Cl−]i), increased CBD, not CBF, at 37 °C; however, it increased both CBD and CBF at 25 °C. Conversely, addition of Cl− channel blockers (5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) and 4-[[4-Oxo-2-thioxo-3-[3-trifluoromethyl]phenyl]-5-thiazolidinylidene]methyl] benzoic acid (CFTR(inh)-172)), which increase [Cl−]i, decreased both CBD and CBF, suggesting that CFTR plays a crucial role for maintaining [Cl−]i in these cells. We speculate that Cl− modulates activities of the molecular motors regulating both CBD and CBF in cHNECs. Moreover, application of the CO2/HCO3−-free solution did not change intracellular pH (pHi), and addition of an inhibitor of carbonic anhydrase (acetazolamide) sustained pHi increase induced by the NH4+ pulse, which transiently increased pHi in the absence of acetazolamide. These results indicate that the cHNEC produces a large amount of CO2, which maintains a constant pHi even under the CO2/HCO3−-free condition.
KeywordsAirway Intracellular Cl− Intracellular pH NH4+ pulse Ciliated human nasal epithelial cell
The authors thank Osaka medical College for renting out the video-microscope equipped with a high speed camera. Experiments were carried out in Kyoto Prefectural University of Medicine (2018–2019) and in Ritsumeikan University (2019).
This work was supported by JSPS KAKENHI to YM (No. JP18H03182), JSPS KAKENHI to MY (No. JP18K09325), and research funding from Saisei Mirai.
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