The human CFTR protein expressed in CHO cells activates aquaporin-3 in a cAMP-dependent pathway: study by digital holographic microscopy

Handle URI:
http://hdl.handle.net/10754/555774
Title:
The human CFTR protein expressed in CHO cells activates aquaporin-3 in a cAMP-dependent pathway: study by digital holographic microscopy
Authors:
Jourdain, P.; Becq, F.; Lengacher, S.; Boinot, C.; Magistretti, Pierre J. ( 0000-0002-6678-320X ) ; Marquet, P.
Abstract:
The transmembrane water movements during cellular processes and their relationship to ionic channel activity remain largely unknown. As an example, in epithelial cells it was proposed that the movement of water could be directly linked to cystic fibrosis transmembrane conductance regulator (CFTR) protein activity through a cAMP-stimulated aqueous pore, or be dependent on aquaporin. Here, we used digital holographic microscopy (DHM) an interferometric technique to quantify in situ the transmembrane water fluxes during the activity of the epithelial chloride channel, CFTR, measured by patch-clamp and iodide efflux techniques. We showed that the water transport measured by DHM is fully inhibited by the selective CFTR blocker CFTRinh172 and is absent in cells lacking CFTR. Of note, in cells expressing the mutated version of CFTR (F508del-CFTR), which mimics the most common genetic alteration encountered in cystic fibrosis, we also show that the water movement is profoundly altered but restored by pharmacological manipulation of F508del-CFTR-defective trafficking. Importantly, whereas activation of this endogenous water channel required a cAMP-dependent stimulation of CFTR, activation of CFTR or F508del-CFTR by two cAMP-independent CFTR activators, genistein and MPB91, failed to trigger water movements. Finally, using a specific small-interfering RNA against the endogenous aquaporin AQP3, the water transport accompanying CFTR activity decreased. We conclude that water fluxes accompanying CFTR activity are linked to AQP3 but not to a cAMP-stimulated aqueous pore in the CFTR protein.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division
Citation:
The human CFTR protein expressed in CHO cells activates aquaporin-3 in a cAMP-dependent pathway: study by digital holographic microscopy 2013, 127 (3):546 Journal of Cell Science
Publisher:
The Company of Biologists
Journal:
Journal of Cell Science
Issue Date:
11-Dec-2013
DOI:
10.1242/jcs.133629
Type:
Article
ISSN:
0021-9533; 1477-9137
Additional Links:
http://jcs.biologists.org/cgi/doi/10.1242/jcs.133629
Appears in Collections:
Articles; Biological and Environmental Sciences and Engineering (BESE) Division; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorJourdain, P.en
dc.contributor.authorBecq, F.en
dc.contributor.authorLengacher, S.en
dc.contributor.authorBoinot, C.en
dc.contributor.authorMagistretti, Pierre J.en
dc.contributor.authorMarquet, P.en
dc.date.accessioned2015-05-26T08:04:24Zen
dc.date.available2015-05-26T08:04:24Zen
dc.date.issued2013-12-11en
dc.identifier.citationThe human CFTR protein expressed in CHO cells activates aquaporin-3 in a cAMP-dependent pathway: study by digital holographic microscopy 2013, 127 (3):546 Journal of Cell Scienceen
dc.identifier.issn0021-9533en
dc.identifier.issn1477-9137en
dc.identifier.doi10.1242/jcs.133629en
dc.identifier.urihttp://hdl.handle.net/10754/555774en
dc.description.abstractThe transmembrane water movements during cellular processes and their relationship to ionic channel activity remain largely unknown. As an example, in epithelial cells it was proposed that the movement of water could be directly linked to cystic fibrosis transmembrane conductance regulator (CFTR) protein activity through a cAMP-stimulated aqueous pore, or be dependent on aquaporin. Here, we used digital holographic microscopy (DHM) an interferometric technique to quantify in situ the transmembrane water fluxes during the activity of the epithelial chloride channel, CFTR, measured by patch-clamp and iodide efflux techniques. We showed that the water transport measured by DHM is fully inhibited by the selective CFTR blocker CFTRinh172 and is absent in cells lacking CFTR. Of note, in cells expressing the mutated version of CFTR (F508del-CFTR), which mimics the most common genetic alteration encountered in cystic fibrosis, we also show that the water movement is profoundly altered but restored by pharmacological manipulation of F508del-CFTR-defective trafficking. Importantly, whereas activation of this endogenous water channel required a cAMP-dependent stimulation of CFTR, activation of CFTR or F508del-CFTR by two cAMP-independent CFTR activators, genistein and MPB91, failed to trigger water movements. Finally, using a specific small-interfering RNA against the endogenous aquaporin AQP3, the water transport accompanying CFTR activity decreased. We conclude that water fluxes accompanying CFTR activity are linked to AQP3 but not to a cAMP-stimulated aqueous pore in the CFTR protein.en
dc.publisherThe Company of Biologistsen
dc.relation.urlhttp://jcs.biologists.org/cgi/doi/10.1242/jcs.133629en
dc.rightsArchived with thanks to Journal of Cell Scienceen
dc.subjectDigital holographic microscopyen
dc.subjectCFTR proteinen
dc.subjectAquaporin AQP3en
dc.subjectWater transporten
dc.subjectcAMP pathwayen
dc.subjectF508del-CFTRen
dc.titleThe human CFTR protein expressed in CHO cells activates aquaporin-3 in a cAMP-dependent pathway: study by digital holographic microscopyen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalJournal of Cell Scienceen
dc.eprint.versionPublisher's Version/PDFen
dc.contributor.institutionBrain Mind Institute, Ecole Polytechnique Fédérale de Lausanne (EPFL), 1015 Lausanne, Switzerlanden
dc.contributor.institutionInstitut de Physiologie et Biologie Cellulaires, CNRS, Université de Poitiers, 1 rue Georges Bonnet, 86022 Poitiers, Franceen
dc.contributor.institutionDepartment of Psychiatry DP-CHUV, Center for Psychiatric Neuroscience, 1008 Prilly-Lausanne, Switzerlanden
kaust.authorMagistretti, Pierre J.en
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