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Chloride transport in apical membrane vesicles from bovine tracheal epithelium: Characterization using a fluorescent indicator

The Journal of Membrane Biology volume 104, pages 233–239 (1988)Cite this article

Summary

Cl transport in apical membrane vesicles derived from bovine tracheal epithelial cells was studied using the Cl-sensitive fluorescent indicator 6-methoxy-N-(3-sulfopropyl) quinolinium. With an inwardly directed 50 mM Cl gradient at 23°C, the initial rate of Cl entry (J Cl) was increased significantly from 0.32±0.12 nmol · sec−1 · mg protein−1 (mean±sem) to 0.50±0.07 nmol · sec−1 · mg protein−1 when membrane potential was changed from 0 to +60 mV with K/valinomycin. At 37°C, with membrane potential clamped at 0 mV, there was a 34±7% (n=5) decrease inJ Cl from a control value of 0.37±0.03 nmol · sec−1 · mg protein−1 upon addition of 0.2mm diphenylamine-2-carboxylate. The following did not alterJ Cl significantly (J Cl values gives as percent change from control): 50mm cis Na (−1±5%), 0.1mm furosemide (−3±4%), 0.1mm furosemide in the presence of 50mm cis Na (−5±2%), 0.1mm H2DIDS (−18±9%), a 1.5 pH unit inwardly directed H gradient (−7±7%), and 0.1mm H2DIDS in the presence of a 1.5 unit pH gradient (4±18%). With inward 50mm anion gradients, the initial rates of Br and I entry (J Br andJ 1, respectively) were not significantly different fromJ Cl.J Cl was a saturable function of Cl concentration with apparentK d of 24mm and apparentV max of 0.54 nmol · sec−1 · mg protein−1. Measurement of the temperature dependence ofJ Cl yielded an activation energy of 5.0 kcal/mol (16–37°C). These results demonstrate that Cl transport in tracheal apical membrane vesicles is voltage-dependent and inhibited by diphenylamine-2-carboxylate. There is no significant contribution from the Na/K/2Cl, Na/Cl, or Cl/OH(H) transporters. The conductive pathway does not discriminate between Cl, Br, and I and is saturable. The low activation energy supports a pore-type mechanism for the conductance.

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Affiliations

  1. Department of Physiology Cardiovascular Research Institute, and Cystic Fibrosis Research Center, University of California, San Francisco, California

    P. Fong & J. H. Widdicombe

  2. Department of Medicine, Cardiovascular Research Institute and Cystic Fibrosis Research Center, University of California, San Francisco, California

    Nicholas P. Illsley & A. S. Verkman

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  1. P. Fong
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  2. Nicholas P. Illsley
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  3. J. H. Widdicombe
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  4. A. S. Verkman
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Fong, P., Illsley, N.P., Widdicombe, J.H. et al. Chloride transport in apical membrane vesicles from bovine tracheal epithelium: Characterization using a fluorescent indicator. J. Membrain Biol. 104, 233–239 (1988). https://doi.org/10.1007/BF01872325

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  • DOI: https://doi.org/10.1007/BF01872325

Key Words

  • trachea
  • ion permeability
  • chloride channel
  • fluorescence