The phorbol ester PMA and cyclic AMP activate different Cl- and HCO3/- fluxes in C127 cells expressing CFTR

Olga Zegarra-Moran, Anna Maria Porcelli, Michela Rugolo

Research output: Contribution to journalArticlepeer-review

Abstract

In mouse mammary epithelial C127 cells expressing wild-type cystic fibrosis transmembrane conductance regulator (CFTR), chloride efflux, measured with the Cl- sensitive dye 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ), was stimulated by activation of protein kinase A with cyclic AMP elevating agents forskolin plus 3-isobutyl-1-methyl-xanthine (IBMX) and, to a less extent, by activation of protein kinase C with the phorbol 12-myristate 13-acetate (PMA). Conversely, bicarbonate influx, determined by intracellular alkalinization of cells incubated with the pH-sensitive dye 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluoresceintetraacetoxym ethyl ester (BCECF-AM), was stimulated by cyclic AMP elevation, but not by PMA. Patch clamp analysis revealed that PMA activated a Cl- current with the typical biophysical characteristics of swelling-activated current and not of CFTR.

Original languageEnglish
Pages (from-to)120-127
Number of pages8
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1535
Issue number2
DOIs
Publication statusPublished - Feb 14 2001

Keywords

  • CFTR
  • Cystic fibrosis
  • HCO/ influx
  • Patch clamp
  • Phorbol ester (PMA)
  • Swelling-activated chloride current

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Biophysics

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