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

Biochim Biophys Acta. 2001 Feb 14;1535(2):120-7. doi: 10.1016/s0925-4439(00)00089-2.

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)-carboxyfluoresceintetraacetoxymethyl 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiporters / metabolism
  • Bicarbonates / metabolism*
  • Biological Transport
  • Cell Line
  • Cell Membrane / metabolism
  • Chloride-Bicarbonate Antiporters
  • Chlorides / metabolism*
  • Cyclic AMP / pharmacology*
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism*
  • Enzyme Activation / drug effects
  • Fluorescent Dyes
  • Mice
  • Patch-Clamp Techniques
  • Protein Kinase C / metabolism
  • Quinolinium Compounds
  • Tetradecanoylphorbol Acetate / pharmacology*

Substances

  • Antiporters
  • Bicarbonates
  • Chloride-Bicarbonate Antiporters
  • Chlorides
  • Fluorescent Dyes
  • Quinolinium Compounds
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • 6-methoxy-N-(3-sulfopropyl)quinolinium
  • Cyclic AMP
  • Cyclic AMP-Dependent Protein Kinases
  • Protein Kinase C
  • Tetradecanoylphorbol Acetate