Properties of CFTR activated by the xanthine derivative X-33 in human airway Calu-3 cells

Laurence Bulteau, Renaud Dérand, Yvette Mettey, Thierry Métayé, M. Rachel Morris, Ceinwen M. McNeilly, Chiara Folli, Luis J V Galietta, Olga Zegarra-Moran, Malcolm M C Pereira, Chantal Jougla, Robert L. Dormer, Jean Michel Vierfond, Michel Joffre, Frédéric Becq

Research output: Contribution to journalArticle

31 Citations (Scopus)

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.

Original languageEnglish
JournalAmerican Journal of Physiology - Cell Physiology
Volume279
Issue number6 48-6
Publication statusPublished - 2000

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Xanthine
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
Derivatives
1-Methyl-3-isobutylxanthine
Glyburide
Short circuit currents
Phosphorylation
Clamping devices
Iodides
Cystic Fibrosis
Proteins
Western Blotting
Chemical activation
Pharmacology
Membranes

Keywords

  • Chloride conductance
  • Cystic fibrosis transmembrane conductance regulator
  • Pharmacology

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Physiology (medical)

Cite this

Bulteau, L., Dérand, R., Mettey, Y., Métayé, T., Morris, M. R., McNeilly, C. M., ... Becq, F. (2000). Properties of CFTR activated by the xanthine derivative X-33 in human airway Calu-3 cells. American Journal of Physiology - Cell Physiology, 279(6 48-6).

Properties of CFTR activated by the xanthine derivative X-33 in human airway Calu-3 cells. / Bulteau, Laurence; Dérand, Renaud; Mettey, Yvette; Métayé, Thierry; Morris, M. Rachel; McNeilly, Ceinwen M.; Folli, Chiara; Galietta, Luis J V; Zegarra-Moran, Olga; Pereira, Malcolm M C; Jougla, Chantal; Dormer, Robert L.; Vierfond, Jean Michel; Joffre, Michel; Becq, Frédéric.

In: American Journal of Physiology - Cell Physiology, Vol. 279, No. 6 48-6, 2000.

Research output: Contribution to journalArticle

Bulteau, L, Dérand, R, Mettey, Y, Métayé, T, Morris, MR, McNeilly, CM, Folli, C, Galietta, LJV, Zegarra-Moran, O, Pereira, MMC, Jougla, C, Dormer, RL, Vierfond, JM, Joffre, M & Becq, F 2000, 'Properties of CFTR activated by the xanthine derivative X-33 in human airway Calu-3 cells', American Journal of Physiology - Cell Physiology, vol. 279, no. 6 48-6.
Bulteau L, Dérand R, Mettey Y, Métayé T, Morris MR, McNeilly CM et al. Properties of CFTR activated by the xanthine derivative X-33 in human airway Calu-3 cells. American Journal of Physiology - Cell Physiology. 2000;279(6 48-6).
Bulteau, Laurence ; Dérand, Renaud ; Mettey, Yvette ; Métayé, Thierry ; Morris, M. Rachel ; McNeilly, Ceinwen M. ; Folli, Chiara ; Galietta, Luis J V ; Zegarra-Moran, Olga ; Pereira, Malcolm M C ; Jougla, Chantal ; Dormer, Robert L. ; Vierfond, Jean Michel ; Joffre, Michel ; Becq, Frédéric. / Properties of CFTR activated by the xanthine derivative X-33 in human airway Calu-3 cells. In: American Journal of Physiology - Cell Physiology. 2000 ; Vol. 279, No. 6 48-6.
@article{d74bc422652a4acaacc413dc83bc604b,
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.",
keywords = "Chloride conductance, Cystic fibrosis transmembrane conductance regulator, Pharmacology",
author = "Laurence Bulteau and Renaud D{\'e}rand and Yvette Mettey and Thierry M{\'e}tay{\'e} and Morris, {M. Rachel} and McNeilly, {Ceinwen M.} and Chiara Folli and Galietta, {Luis J V} and Olga Zegarra-Moran and Pereira, {Malcolm M C} and Chantal Jougla and Dormer, {Robert L.} and Vierfond, {Jean Michel} and Michel Joffre and Fr{\'e}d{\'e}ric Becq",
year = "2000",
language = "English",
volume = "279",
journal = "American Journal of Physiology",
issn = "0363-6119",
publisher = "American Physiological Society",
number = "6 48-6",

}

TY - JOUR

T1 - Properties of CFTR activated by the xanthine derivative X-33 in human airway Calu-3 cells

AU - Bulteau, Laurence

AU - Dérand, Renaud

AU - Mettey, Yvette

AU - Métayé, Thierry

AU - Morris, M. Rachel

AU - McNeilly, Ceinwen M.

AU - Folli, Chiara

AU - Galietta, Luis J V

AU - Zegarra-Moran, Olga

AU - Pereira, Malcolm M C

AU - Jougla, Chantal

AU - Dormer, Robert L.

AU - Vierfond, Jean Michel

AU - Joffre, Michel

AU - Becq, Frédéric

PY - 2000

Y1 - 2000

N2 - 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.

AB - 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.

KW - Chloride conductance

KW - Cystic fibrosis transmembrane conductance regulator

KW - Pharmacology

UR - http://www.scopus.com/inward/record.url?scp=0033638908&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033638908&partnerID=8YFLogxK

M3 - Article

C2 - 11078708

AN - SCOPUS:0033638908

VL - 279

JO - American Journal of Physiology

JF - American Journal of Physiology

SN - 0363-6119

IS - 6 48-6

ER -