title: Functional coupling of TRPV4 cationic channel and large conductance, calcium-dependent potassium channel in human bronchial epithelial cell lines.
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 (BK(Ca)) 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 Ca(2+) entry, and outwardly rectifying cationic currents elicited by the TRPV4 specific activator 4alpha-phorbol 12,13-didecanoate (4alphaPDD). Cell-attached and whole-cell patch-clamp of HBE cells exposed to 4alphaPDD, and hypotonic and high-viscosity solutions (related to mechanical stress) revealed the activation of BK(Ca) channels subsequent to extracellular Ca(2+) influx via TRPV4, an effect lost upon antisense-mediated knock-down of TRPV4. Further analysis of BK(Ca) modulation after TRPV4 activation showed that the Ca(2+) signal can be generated away from the BK(Ca) location at the plasma membrane, and it is not mediated by intracellular Ca(2+) release via ryanodine receptors. Finally, we have shown that, unlike the reported disengagement of TRPV4 and BK(Ca) in response to hypotonic solutions, cystic fibrosis bronchial epithelial cells (CFBE) preserve the functional coupling of TRPV4 and BK(Ca) in response to high-viscous solutions.
authors.affiliation: Department of Experimental and Health Sciences, Universitat Pompeu Fabra, PRBB, C/ Dr. Aiguader 88, Barcelona, 08003, Spain.
authors.forename: José M
authors.initials: JM
authors.lastname: Fernández-Fernández
authors.forename: Yaniré N
authors.initials: YN
authors.lastname: Andrade
authors.forename: Maite
authors.initials: M
authors.lastname: Arniges
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Fernandes
authors.forename: Cristina
authors.initials: C
authors.lastname: Plata
authors.forename: Francisca
authors.initials: F
authors.lastname: Rubio-Moscardo
authors.forename: Esther
authors.initials: E
authors.lastname: Vázquez
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Valverde
mesh_terms: D001980:Bronchi / Q000166:cytology / Q000502:physiology*; D002460:Cell Line; D003550:Cystic Fibrosis / Q000473:pathology; D004594:Electrophysiology; D004847:Epithelial Cells / Q000502:physiology*; D006801:Humans; D007038:Hypotonic Solutions / Q000494:pharmacology; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000502:physiology*; D020320:Oligoribonucleotides, Antisense / Q000494:pharmacology; D009997:Osmotic Pressure; D010703:Phorbol Esters / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050916:TRPV Cation Channels / Q000502:physiology*; D014783:Viscosity
chemical_list: D007038:Hypotonic Solutions; D051036:Large-Conductance Calcium-Activated Potassium Channels; D020320:Oligoribonucleotides, Antisense; D010703:Phorbol Esters; D050916:TRPV Cation Channels; C416986:TRPV4 protein, human; C008876:phorbol-12,13-didecanoate