title: Properties of CFTR activated by the xanthine derivative X-33 in human airway Calu-3 cells.
abstract: The pharmacological activation of the cystic fibrosis gene protein cystic fibrosis transmembrane conductance regulator (CFTR) was studied in human airway epithelial Calu-3 cells, which express a high level of CFTR protein as assessed by Western blot and in vitro phosphorylation. Immunolocalization shows that CFTR is located in the apical membrane. We performed iodide efflux, whole cell patch-clamp, and short-circuit recordings to demonstrate that the novel synthesized xanthine derivative 3, 7-dimethyl-1-isobutylxanthine (X-33) is an activator of the CFTR channel in Calu-3 cells. Whole cell current activated by X-33 or IBMX is linear, inhibited by glibenclamide and diphenylamine-2-carboxylate but not by DIDS or TS-TM calix[4]arene. Intracellular cAMP was not affected by X-33. An outwardly rectifying Cl(-) current was recorded in the absence of cAMP and X-33 stimulation, inhibited by DIDS and TS-TM calix[4]arene. With the use of short-circuit recordings, X-33 and IBMX were able to stimulate a large concentration-dependent CFTR transport that was blocked by glibenclamide but not by DIDS. Our results show that manipulating the chemical structure of xanthine derivatives offers an opportunity to identify further specific activators of CFTR in airway cells.
authors.affiliation: Laboratoire de Physiologie des Régulations Cellulaires, Unité Mixte de Recherche 6558, 86022 Poitiers, France.
authors.forename: L
authors.initials: L
authors.lastname: Bulteau
authors.forename: R
authors.initials: R
authors.lastname: Dérand
authors.forename: Y
authors.initials: Y
authors.lastname: Mettey
authors.forename: T
authors.initials: T
authors.lastname: Métayé
authors.forename: M R
authors.initials: MR
authors.lastname: Morris
authors.forename: C M
authors.initials: CM
authors.lastname: McNeilly
authors.forename: C
authors.initials: C
authors.lastname: Folli
authors.forename: L J
authors.initials: LJ
authors.lastname: Galietta
authors.forename: O
authors.initials: O
authors.lastname: Zegarra-Moran
authors.forename: M M
authors.initials: MM
authors.lastname: Pereira
authors.forename: C
authors.initials: C
authors.lastname: Jougla
authors.forename: R L
authors.initials: RL
authors.lastname: Dormer
authors.forename: J M
authors.initials: JM
authors.lastname: Vierfond
authors.forename: M
authors.initials: M
authors.lastname: Joffre
authors.forename: F
authors.initials: F
authors.lastname: Becq
mesh_terms: D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D016466:CHO Cells; D002110:Caffeine / Q000494:pharmacology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002712:Chlorides / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005905:Glyburide / Q000494:pharmacology; D007004:Hypoglycemic Agents / Q000494:pharmacology; D066298:In Vitro Techniques; D007454:Iodides / Q000493:pharmacokinetics; D007457:Iodine Radioisotopes; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism; D018408:Patch-Clamp Techniques; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010766:Phosphorylation; D020545:Respiratory Mucosa / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D013806:Theophylline / Q000031:analogs & derivatives* / Q000494:pharmacology; D014970:Xanthines / Q000138:chemical synthesis / Q000494:pharmacology*; D062367:ortho-Aminobenzoates / Q000494:pharmacology
chemical_list: D002121:Calcium Channel Blockers; D002712:Chlorides; D007004:Hypoglycemic Agents; D007454:Iodides; D007457:Iodine Radioisotopes; D008565:Membrane Proteins; D010726:Phosphodiesterase Inhibitors; C417199:X 33 compound; D014970:Xanthines; D062367:ortho-Aminobenzoates; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005576:Colforsin; C053907:8-cyclopentyl-1,3-dimethylxanthine; D002110:Caffeine; C001209:fenamic acid; D013806:Theophylline; D000242:Cyclic AMP; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D005905:Glyburide; D015056:1-Methyl-3-isobutylxanthine