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Functional coupling of TRPV4 cationic channel and large conductance, calcium-dependent potassium channel in human bronchial epithelial cell lines

  • Ion Channels, Receptors and Transporters
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Abstract

Calcium-dependent potassium channels are implicated in electrolyte transport, cell volume regulation and mechanical responses in epithelia, although the pathways for calcium entry and their coupling to the activation of potassium channels are not fully understood. We now show molecular evidence for the presence of TRPV4, a calcium permeable channel sensitive to osmotic and mechanical stress, and its functional coupling to the large conductance calcium-dependent potassium channel (BKCa) in a human bronchial epithelial cell line (HBE). Reverse transcriptase polymerase chain reaction, intracellular calcium imaging and whole-cell patch–clamp experiments using HBE cells demonstrated the presence of TRPV4 messenger and Ca2+ entry, and outwardly rectifying cationic currents elicited by the TRPV4 specific activator 4α-phorbol 12,13-didecanoate (4αPDD). Cell-attached and whole-cell patch–clamp of HBE cells exposed to 4αPDD, and hypotonic and high-viscosity solutions (related to mechanical stress) revealed the activation of BKCa channels subsequent to extracellular Ca2+ influx via TRPV4, an effect lost upon antisense-mediated knock-down of TRPV4. Further analysis of BKCa modulation after TRPV4 activation showed that the Ca2+ signal can be generated away from the BKCa location at the plasma membrane, and it is not mediated by intracellular Ca2+ release via ryanodine receptors. Finally, we have shown that, unlike the reported disengagement of TRPV4 and BKCa in response to hypotonic solutions, cystic fibrosis bronchial epithelial cells (CFBE) preserve the functional coupling of TRPV4 and BKCa in response to high-viscous solutions.

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Acknowledgment

We thank D. C. Gruenert (San Francisco, USA) for the gift of the cell lines. Work funded by the Spanish Ministry of Education and Science (SAF2006-04973 to M.A.V. and SAF2006-13893-C02-02 to J.M.F.-F.), Ministry of Health (Fondo de Investigación Sanitaria, red HERACLES RD06/0009 to M.A.V.), Generalitat de Catalunya (SGR05-266 to M.A.V.), and Fundació la Marató de TV3 (061331 to J.M.F.-F.). J.M.F.-F. is a Ramón y Cajal Fellow. The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Laboratory of Molecular Physiology and Channelopathies, Department of Experimental and Health Sciences, Universitat Pompeu Fabra, PRBB, C/ Dr. Aiguader 88, Barcelona, 08003, Spain

    José M. Fernández-Fernández, Yaniré N. Andrade, Maite Arniges, Jacqueline Fernandes, Cristina Plata, Francisca Rubio-Moscardo, Esther Vázquez & Miguel A. Valverde

  2. Laboratory of Molecular Physiology and Channelopathies, Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Parc de Recerca Biomedica de Barcelona, C/ Dr. Aiguader 88, Barcelona, 08003, Spain

    Miguel A. Valverde

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  1. José M. Fernández-Fernández
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Correspondence to Miguel A. Valverde.

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José M. Fernández-Fernández and Yaniré N. Andrade contributed equally to this work, and both should be considered first authors.

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Fernández-Fernández, J.M., Andrade, Y.N., Arniges, M. et al. Functional coupling of TRPV4 cationic channel and large conductance, calcium-dependent potassium channel in human bronchial epithelial cell lines. Pflugers Arch - Eur J Physiol 457, 149–159 (2008). https://doi.org/10.1007/s00424-008-0516-3

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