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Pflügers Archiv

, Volume 422, Issue 6, pp 539–545 | Cite as

Regulation of chloride channels in the human colon carcinoma cell line HT29.cl19A

  • Maarten Kansen
  • Rajesh B. Bajnath
  • Jack A. Groot
  • Hugo R. de Jonge
  • Bob Scholte
  • André T. Hoogeveen
  • Jan Bijman
Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands

Abstract

Chloride (Cl) channels are important in the regulation of salt and water transport in secretory epithelial cells. A disturbed Cl secretion is the most consistent characteristic in the genetic disease cystic fibrosis. An outwardly rectifying Cl channel (OR) with a conductance of 25–50 pS had been proposed to play a major role in Cl secretion. Activation by Ca2+ and the protein kinases (PK) A and C (at less than 10 nM Ca2+) as well as inhibition by PKC (at 1 μM Ca2+) has been reported. In the present study, we have identified and characterized the OR in HT29.cl19A human colon carcinoma cells. The OR displayed a conductance of 31±4 pS (n=25). Its open probability in 10 nM Ca2+ was voltage-dependent in 50% of the patches, starting from 0.2 at -70 mV to 0.8 at 70 mV. The spontaneous activation in excised inside-out patches at −60 mV was Ca2+-dependent and decreased from 29% in 1 mM Ca2+ to 2% in 10 nM Ca2+. Active OR were found in (a) 25% of patches exposed to 10 nM Ca2+, ATP and cAMP only, (b) 42% of the patches exposed to 10 nM Ca2+, ATP and the catalytic subunit of PKA (CAK) and (c) 67% of the patches exposed to 1 mM Ca2+, ATP plus CAK. Inhibition of voltage-activated channels by addition of PKC in 1 μM or 1 mM Ca2+ was not observed. Attempts to activate the OR in cell-attached patches by increasing cAMP levels under different experimental conditions were unsuccessful. Our data suggest that the OR may not be as important in Cl secretion as has been thought.

Key words

Chloride channel Patch clamp Intestine 

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Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Maarten Kansen
    • 1
  • Rajesh B. Bajnath
    • 3
  • Jack A. Groot
    • 3
  • Hugo R. de Jonge
    • 2
  • Bob Scholte
    • 1
  • André T. Hoogeveen
    • 1
  • Jan Bijman
    • 1
  1. 1.Department of Cell Biology and Genetics, Medical FacultyErasmus University RotterdamRotterdamThe Netherlands
  2. 2.Department of Biochemistry I, Medical FacultyErasmus University RotterdamRotterdamThe Netherlands
  3. 3.Department of Experimental ZoologyUniversity of AmsterdamAmsterdamThe Netherlands

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