Abstract
The single channel inside-out patch clamp technique was used to characterize ion channels in the apical membranes of ragweed-sensitized and control canine tracheal epithelial cells maintained in primary culture. Patches were obtained from single isolated cells or from cells at the edges of confluent sheets. A new type of chloride channel was seen in sensitized cells but not in control cells. The channel showed inward rectification in symmetric chloride solutions with conductance varying from 95 pS to 52 pS over the range of −60 mV to 60 mV membrane potential. Channel gating was voltage dependent with maximal opening at about −30 mV Kinetic analysis showed that distributions of closed and open times could both be well fitted by the sums of three exponential components. Rate constants for transitions between the states of a linear kinetic model were calculated, with only one rate being significantly voltage dependent. The possible significance of this channel is discussed.
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Becker AB, Hershkovich J, Simons KJ, Lilley MK, Simons FER, Kepron MW (1989) Development of chronic airway hyperresponsiveness in ragweed-sensitized allergic dogs. J Appl Physiol 66:2691–2697
Colquhoun D, Hawkes AG (1977) Relaxation and fluctuations of membrane currents that flow through drug-operated ion channels. Proc R Soc Lond B199:231–262
Colquhoun D, Hawkes AG (1981) On the stochastic properties of single ion channels. Proc R Soc Lond B211:205–235
Colquhoun D, Sigworth FJ (1983) Fitting and statistical analysis of single-channel records. In: Sakmann B, Neher E (eds) Single-channel recording. Plenum Press, New York, pp 191–263
Duszyk M, French AS, Man SFP (1989) Cystic fibrosis affects chloride and sodium channels in human airway epithelia. Can J Physiol Pharmacol 67:1362–1365
Frizzell RA (1987) Cystic fibrosis: a disease of ionic channels? TINS 10:190–195
Frizzell RA, Halm DR, Rechkemmer G, Shoemaker RL (1986) Chloride channel regulation in secretory epithelia. Fed Proc 45:2727–2731
Gögelein H (1988) Chloride channels in epithelia. Biochim Biophys Acta 947:521–547
Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high resolution current recording from cells and cell-free membrane patches. Pflügers Arch 391:85–100
Hanrahan JW, Alles WP, Lewis SA (1985) Single anion-selective channels in basolateral membrane of a mammalian tight epithelium Proc Natl Acad Sci USA 82:7791–7795
Horn R (1987) Statistical methods for model discrimination. Applications to gating kinetics and permeation of the acetylcholine receptor channel. Biophys J 51:255–263
Korn SJ, Horn R (1988) Statistical discrimination of fractal and Markov models of single-channel gating. Biophys J 54:871–877
Kunzelmann K, Pavenstädt H, Beck C, Ünal Ö, Emmerich P, Arndt HJ, Greger R (1989) Characterization of potassium channels in respiratory cells. I. General properties. Pflügers Arch 414:291–296
Man SFP, Duszyk M, French AS (1989) Sodium channels in the apical membrane of human airway epithelial cells. Am Rev Respir Dis 139:A477, 1989
Shoemaker RL, Frizzell RA, Dwyer TM, Farley JM (1986) Single chloride channel currents from canine tracheal cells. Biochim Biophys Acta 858:235–242
Sigworth FJ, Sine S (1987) Data transformation for improved display and fitting of single-channel dwell time histograms. Biophys J 52:1047–1054
Wu R, Yankaskas J, Cheng E, Knowles MR, Boucher RC (1985) Growth and differentiation of human nasal epithelial cells in culture: serum free, hormone-supplemented medium and proteoglycan synthesis. Am Rev Respir Dis 132:311–320
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Duszyk, M., French, A.S., Man, S.F.P. et al. An inwardly rectifying chloride channel in ragweed-sensitized canine tracheal epithelial cells. Eur Biophys J 20, 65–69 (1991). https://doi.org/10.1007/BF00186254
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DOI: https://doi.org/10.1007/BF00186254