Both CFTR and outwardly rectifying chloride channels contribute to cAMP- stimulated whole cell chloride currents

E. M. Schwiebert, T. Flotte, G. R. Cutting, W. B. Guggino

Research output: Contribution to journalArticlepeer-review

Abstract

From whole cell patch-clamp recordings at 35°C utilizing either nystatin perforation or conventional methods with 5 mM MgATP in the pipette solution, it was demonstrated that both cystic fibrosis transmembrane conductance regulator (CFTR) chloride (Cl-) channels and outwardly rectifying Cl- channels (ORCC) contribute to adenosine 3',5'-cyclic monophosphate (cAMP)- activated whole cell Cl- currents in cultured human airway epithelial cells. These results were similar whether recordings were performed on two normal human cell lines or on two cystic fibrosis (CF) cell lines stably complemented with wild-type CF gene. These results were obtained by exploiting dissimilar biophysical properties of CFTR and ORCC currents such as the degree of rectification of the current-voltage relationship, the difference in sensitivity to Cl- channel-blocking drugs such as 4,4'- diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), calixarenes, and diphenylamine carboxylic acid (DPC), and the opposing Cl- relative to I- permeabilities of the two channels. In normal cells or complemented CF cells, 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate stimulated outwardly rectifying whole cell Cl- currents. Addition of DIDS in the presence of cAMP inhibited the outwardly rectifying portion of the cAMP- activated Cl- current. The remaining cAMP-activated, DIDS-insensitive, linear CFTR Cl- current was inhibited completely by DPC. Additional results showed that not only do ORCC and CFTR Cl- channels contribute to cAMP- activated Cl- currents in airway epithelial cells where wild-type CFTR is expressed but that both channels fail to respond to cAMP in ΔF508-CFTR- containing CF airway cells. We conclude that CFTR not only functions as a cAMP-regulated Cl- channel in airway epithelial cells but also controls the regulation of ORCC.

Original languageEnglish (US)
Pages (from-to)C1464-C1477
JournalAmerican Journal of Physiology - Cell Physiology
Volume266
Issue number5 35-5
DOIs
StatePublished - 1994

Keywords

  • adeno-associated viral vectors
  • adenosine 3',5'-cyclic monophosphate
  • airway epithelia
  • cell culture
  • cystic fibrosis
  • cystic fibrosis transmembrane conductance regulator
  • gene transfection
  • patch clamp

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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