Skip to main content
SpringerLink
Log in

Properties and regulation of chloride channels in cystic fibrosis and normal airway cells

  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

The present study examines the properties of Cl−channels in cultured respiratory cells of cystic fibrosis (CF) patients and normal (N) individuals. In excised membrane patches the conductances for CF and N Cl− channels were larger at positive as compared to negative clamp voltages (V c): 74±2.6 (V c > 0) and 47±2.0 pS (V c < 0) for CF (n= 57) and 69±3.6 (V c > 0) and 45±2.3 pS (V c < 0) for N (n=35). The open probability (P o) of the channel increased markedly with depolarization. Both the voltage dependence of the conductance and of P o contribute to the outward rectification of the channel. The time histogram analysis reveals two open and two closed time constants. The selectivity of the channel was Cl−=Br− =I− > NO −3 ≫ gluconate. The channel was inhibited reversibly by 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB) at 10−7 mol/l to 10−5 mol/l. While Cl− channels were present in cell attached patches of N cells, they were absent in those of CF cells. The mean conductance for cell attached (N) Cl− channels was 76±3.2 pS for positive clamp voltages (V c) and 46±3.9 pS for negative V c (n=8). When the membrane patches were excised from CF cells Cl− currents appeared spontaneously (n=19). The immediate appearance (within 1 s) of Cl− channels after excision was observed at positive (n=6) as well as at negative clamp voltage (n=13). “Excision activation” of CF Cl− channels was observed at low (< 10−9 mol/l) or high (10−3 mol/l) calcium activities on the cytosolic side of the excised patch. Variation of the Ca+ activity (< 10−9–10−3 mol/l) or pH (6.5–8.5) on the cytosolic side exerted no effects on these Cl− channels. These results suggest that Cl− channels are present in the apical membrane of CF and N respiratory cells but they seem to be inhibited in intact CF cells. Excision of the patch and hence removal of the cytosolic “inhibitor” leads to an activation of Cl− channels. The Cl− channels in excised patches of N and CF cells have identical properties.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Argent BE, Arkle S, Gray MA, Greenwell JR (1987) Two types of calcium-sensitive channels in isolated rat pancreatic duct cells. J Physiol 386:82P

    Google Scholar 

  2. Barthelson RA, Jacoby DB, Widdicombe JH (1987) Regulation of chloride secretion in dog tracheal epithelium by protein kinase C. Am J Physiol 253:C802-C808

    Google Scholar 

  3. Baxter PS, Goldhill J, Hardcastle J, Hardcastle PT, Taylor CJ (1988) Intestinal secretion in cystic fibrosis. Disorders of chloride channel regulation: Cystic fibrosis and secretory diarrhea. Koninklijke Nederlandse Academie van Wetenschappen, Amsterdam

    Google Scholar 

  4. Berschneider HM, Knowles MR, Azizkhan RG, Boucher RC, Tobey NA, Orlando RC, Powell DW (1988) Altered intestinal chloride transport in cystic fibrosis. FASEB J 2:2625–2629

    Google Scholar 

  5. Bijman J, Frömter E (1986) Direct demonstration of high transepithelial chloride-conductance in normal human sweat duct which is absent in cystic fibrosis. Pflügers Arch 407:123–127

    Google Scholar 

  6. Bijman J, Kansen M, Hoogeveen AH, Scholte B, Van der Kamp A, De Jonge H (1988) Electrolyte transport in normal and CF epithelia. In: Wong PYD, Young JA (eds) Exocrine secretion. Hong Kong University Press, pp 17–19

  7. Bijman J, Schölte B, De Jonge HR, Hoogeveen AT, Kansen M, Sinaasappel M, van der Kamp AWM (1988) Chloride transport in cystic fibrosis: chloride channel regulation in cultured sweat duct and cultured nasal epithelium. In: Mastella G, Quinton PM. Cellular and molecular basis of cystic fibrosis, San Francisco Press, San Francisco, pp 133–139

    Google Scholar 

  8. Boucher RC, Larsen EH (1988) Comparison of ion transport by cultured secretory and absorptive canine airway epithelia. Am J Physiol 254:C535-C547

    Google Scholar 

  9. Boucher RC, Knowles MR, Stutts MJ, Gatzy JT (1983) Epithelial dysfunction in cystic fibrosis lung disease. Lung 161:1–17

    Google Scholar 

  10. Candia OA, Montoreano R, Podos SM (1977) Effects of the ionophore A23187 on chloride transport across isolated frog cornea. Am J Physiol 233:F94-F101

    Google Scholar 

  11. Cartwright CA, McRoberts JA, Mandel KG, Dharmsathaphorn K (1985) Synergistic action of cyclic adenosine monophosphate- and calcium-mediated chloride secretion in a colonic epithelial cell line. J Clin Invest 76:1837–1842

    Google Scholar 

  12. Chase H, Wong S (1988) Simultaneous measurements of intracellular free [Ca] and Cl secretion in T84 cells reveal Ca's role in the action of carbachol. J Gen Physiol 42:25a

    Google Scholar 

  13. Colquhoun D, Hawkes AG (1983) The principles of the stochastic interpretation of ion channel mechanism. In: Sakmann B, Neher E (eds) Single channel recording. Plenum Press, New York, pp 135–175

    Google Scholar 

  14. Dreinhöfer J, Gögelein H, Greger R (1988) Blocking kinetics of Cl− channels in colonic carcinoma cells (HT29) as revealed by 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB). Biochim Biophys Acta 956:135–142

    Google Scholar 

  15. Evans MG, Marty A, Tan YP, Trautmann A (1986) Blockage of Ca-activated Cl conductance by furosemide in rat lacrimal glands. Pflügers Arch 406:65–68

    Google Scholar 

  16. Findlay I, Petersen OH (1985) Acetylcholine stimulates a Ca2+-dependent Cl− conductance in mouse lacrimal acinar cells. Pflügers Arch 403:328–330

    Google Scholar 

  17. Frizzell RA (1987) Cystic fibrosis: a disease of ion channels? TINS 10:190–193

    Google Scholar 

  18. Frizzell RA, Field M, Schultz SG (1979) Sodium-coupled chloride transport by epithelial tissues. Am J Physiol 236:F1-F8

    Google Scholar 

  19. Frizzell RA, Rechkammer GR, Shoemaker RL (1986) Altered regulation of airway epithelial cell chloride channels in cystic fibrosis. Science 233:558–560

    Google Scholar 

  20. Frizzell RA, Schoumacher RA, Shoemaker RL, Halm DR (1987) Chloride channel regulation in secretory epithelial cells. Pediatr Pulmonol 1 (Suppl):24–25

    Google Scholar 

  21. Frömter E, Disser J (1988) Properties of Na+ channels and Cl−channels in sweat duct and nasal polyp epithelia. Disorders of chloride channel regulation: Cystic fibrosis and secretory diarrhea. Koninklijke Nederlandse Academie van Wetenschappen, Amsterdam

    Google Scholar 

  22. Fulford GR, Blake JR (1986) Mucociliary transport in the lung. J Theor Biol 121:381–402

    Google Scholar 

  23. Gögelein H (1988) Chloride channels in epithelia. Biochim Biophys Acta 947:521–547

    Google Scholar 

  24. Gögelein H, Greger R (1984) Single channel recordings from the basolateral and apical membranes of renal proximal tubules. Pflügers Arch 401:424–426

    Google Scholar 

  25. Gögelein H, Greger R (1986) A voltage-dependent ionic channel in the basolateral membrane of late proximal tubules of the rabbit kidney. Pflügers Arch 407:142–148

    Google Scholar 

  26. Gögelein H, Greger R, Schlatter E (1987) Potassium channels in the basolateral membrane of the rectal gland of squalus acanthias. Regulation and inhibitors. Pflügers Arch 409:107–113

    Google Scholar 

  27. Goldstein JL, Nash NT, Al-Bazzaz F, Layden TJ, Rao MC (1988) Rectum has abnormal ion transport but normal cAMP-binding proteins in cystic fibrosis. Am J Physiol 254:C719-C724

    Google Scholar 

  28. Greger R, Kunzelmann K (1988) Chloride channels. Disorders of chloride channel regulation: Cystic fibrosis and secretory diarrhea. Koninklijke Nederlandse Academie van Wetenschappen, Amsterdam

    Google Scholar 

  29. Greger R, Schlatter E (1984) Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias). I. Experiments in isolated in vitro perfused rectal gland tubules. Pflügers Arch 402:63–75

    Google Scholar 

  30. Greger R, Schlatter E (1984) Mechanism of NaCl secretion in rectal gland tubules of spiny dogfish (Squalus acanthias). II. Effects of inhibitors. Pflügers Arch 402:364–375

    Google Scholar 

  31. Greger R, Gögelein H, Schlatter E (1987) Potassium channels in the basolateral membrane of the rectal gland of the dogfish (Squalus acanthias). Pflügers Arch 409:100–106

    Google Scholar 

  32. Greger R, Schlatter E, Gögelein H (1987) Chloride channels in the luminal membrane of the rectal gland of the dogfish (Squalus acanthias). Properties of the “larger” conductance channel. Pflügers Arch 409:114–121

    Google Scholar 

  33. Halm DR, Rechkemmer GR, Schoumacher RA, Frizzell RA (1988) Apical membrane chloride channels in a colonic cell line activated by secretory agonists. Am J Physiol 254:C505-C 511

    Google Scholar 

  34. 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

    Google Scholar 

  35. Hayslett JP, Gögelein H, Kunzelmann K, Greger R (1987) Characteristics of apical chloride channels in human colon cells (HT29). Pflügers Arch 410:487–494

    Google Scholar 

  36. Knowles MR, Gatzy JT, Boucher RC (1983) Relative ion permeability of normal and cystic fibrosis nasal epithelium. J Clin Invest 71:1410–1417

    Google Scholar 

  37. Kunzelmann K, Ünal Ö, Beck C, Emmrich P, Arndt HJ, Greger R (1988) Ion channels of cultured respiratory epithelial cells of patients with cystic fibrosis. Pflügers Arch 411:R68

    Google Scholar 

  38. Kunzelmann K, Pavenstädt H, Beck C, Ünal Ö, Emmrich P, Arndt HJ, Greger R (1989) Characterization of potassium channels in respiratory cells. I. General properties. Pflügers Arch 414:291–296

    Google Scholar 

  39. Kunzelmann K, Pavenstädt H, Greger R (1989) Characterization of potassium channels in respiratory cells. II. Inhibitors and Regulation. Pflügers Arch 414:297–303

    Google Scholar 

  40. Li M, McCann JD, Liedtke CM, Nairn AC, Greengard P, Welsh MJ (1988) Cyclic AMP-dependent protein kinase opens chloride channels in normal but not cystic fibrosis airway epithelium. Nature 331:358–360

    Google Scholar 

  41. Marin MG, Zaremba MM (1978) Effect of calcium on ion transport and electrical properties of tracheal epithelium. J Appl Physiol 44:900–904

    Google Scholar 

  42. Miller C, White MH (1984) Dimeric structure of single chloride channels from Torpedo electroplax. Proc Natl Acad Sci USA 81:2772–2775

    Google Scholar 

  43. Olver RE, Davis B, Marin MG, Nadel JA (1975) Active transport of Na+ and Cl− across the canine tracheal epithelium in vitro. Am Rev Respir Dis 112:811–815

    Google Scholar 

  44. Orlando RC, Powell DW, Boucher RC, Knowles MR (1985) Colonic and esophageal potential difference measurements in cystic fibrosis. Gastroenterology 88:1524

    Google Scholar 

  45. Quinton PM (1983) Chloride impermeability in cystic fibrosis. Nature 301:421–422

    Google Scholar 

  46. Quinton PM (1986) Missing Cl conductance in cystic fibrosis. Am J Physiol 251:C649-C652

    Google Scholar 

  47. Rohlicek V, Fröbe U, Gögelein H, Greger R (1989) Versatile supplement device with remote control for the control of patch clamp experiments. Pflügers Arch 413:444–446

    Google Scholar 

  48. Sakmann B, Neher E (1984) Patch clamp techniques for studying ionic channels in excitable membranes. Annu Rev Physiol 46:455–472

    Google Scholar 

  49. Sato K, Sato F (1981) Role of calcium in cholinergic and adrenergic mechanisms of eccrine sweat secretion. Am J Physiol 241:C113-C120

    Google Scholar 

  50. Schoumacher RA, Shoemaker RL, Halm DR, Tallant EA, Wallace RW, Frizzell RA (1987) Phosphorylation fails to activate chloride channels from cystic fibrosis airway cells. Nature 330:752–754

    Google Scholar 

  51. Smith PL, Welch MJ, Stoff JS, Frizzell RA (1982) Chloride secretion by canine tracheal epithelium. I. Role of intracellular cAMP levels. J Membr Biol 70:217–226

    Google Scholar 

  52. Wangemann P, Di Stefano A, Wittner M, Englert HC, Lang HJ, Schlatter E, Greger R (1986) Cl−-channel blockers in the thick ascending limb of the loop of Henle. Structure activity relationship. Pflügers Arch 407:S128-S141

    Google Scholar 

  53. Welsh MJ (1986) An apical-membrane chloride channel in human tracheal epithelium. Science 232:1648–1649

    Google Scholar 

  54. Welsh MJ (1987) Electrolyte transport by airway epithelia. Physiol Rev 67:1143–1184

    Google Scholar 

  55. Welsh MJ (1987) Effect of phorbol ester and calcium ionophore on chloride secretion in canine tracheal epithelium. Am J Physiol 253:C828-C834

    Google Scholar 

  56. Welsh MJ, Liedtke CM (1986) Chloride and potassium channels in cystic fibrosis airway epithelia. Nature 322:467–470

    Google Scholar 

  57. Welsh MJ, Mc Cann JD, Dearborn DG (1987) Chloride and sodium channel currents in normal and cystic fibrosis sweat duct epithelium. Fed Proc 46:5567

    Google Scholar 

  58. Widdicombe JH (1986) Cystic fibrosis and β-adrenergic response of airway epithelial cell cultures. Am J Physiol 251:R818-R822

    Google Scholar 

  59. Widdicombe JH, Welsh MJ, Finkbeiner WE (1985) Cystic fibrosis decreases the apical membrane chloride permeability of monolayers cultured from cells of tracheal epithelium. Proc Natl Acad Sci USA 82:6167–6171

    Google Scholar 

  60. Yankaskas JR, Cooton CU, Knowles MR, Gatzy JT (1985) Culture of human nasal epithelium cells on collagen matrix supports. Am Rev Respir Dis 132:1281–1287

    Google Scholar 

  61. Yankaskas JR, Gatzy JT, Knowles MR, Boucher RC (1985) Persistence of abnormal chloride ion permeability in cystic fibrosis nasal epithelial cells in heterologous culture. Lancet 1:954–956

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Physiologisches Institut der Albert-Ludwigs-Universität, Hermann-Herder-Strasse 7, D-7800, Freiburg, Federal Republic of Germany

    Karl Kunzelmann, Hermann Pavenstädt & Rainer Greger

Authors
  1. Karl Kunzelmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hermann Pavenstädt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rainer Greger
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kunzelmann, K., Pavenstädt, H. & Greger, R. Properties and regulation of chloride channels in cystic fibrosis and normal airway cells. Pflügers Arch 415, 172–182 (1989). https://doi.org/10.1007/BF00370589

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00370589

Key words

Search

Navigation