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

  • Zegarra-Moran O
  • Porcelli A
  • Rugolo M
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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. © 2001 Elsevier Science B.V.

Author-supplied keywords

  • CFTR
  • Cystic fibrosis
  • HCO3/-influx
  • Patch clamp
  • Phorbol ester (PMA)
  • Swelling-activated chloride current

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