Skip to main content
SpringerLink
Log in

Chloride transport in cultured nasal epithelium of cystic fibrosis patients

  • Published:
Pflügers Archiv Aims and scope Submit manuscript

Abstract

In this study, nasal polyp epithelial cells from control and cystic fubrosis (CF) patients were cultured using a method which allows multiple passages. The cells were tested in Ussing chamber experiments to study transcellular ion transport. Cultured CF nasal polyp cells did not exhibit spontaneous transcellular chloride transport in the presence of amiloride, in contrast to normal cells. Forskolin increased the short circuit current (I sc) in control but not CF cells. Forskolin and isoproterenol increased the cAMP levels in control and CF cells. Histamine, bradykinin and isoproterenol transiently increased the intracellular calcium level and caused a parallel increase of the transcellular chloride current in both normal and CF cells. The transient effects of isoproterenol were not sensitive to the beta blocker atenol and could not be mimicked by forskolin. We conclude that in cultured nasal polyp cells a difference in chloride transport activity between CF and control cells is retained following multiple passages. Our results suggest that the active state of chloride channels in nasal polyp cells does not require activation of a second messenger pathway. This apparently spontaneous activity appears to be reduced in CF cells. The calcium- but not the cAMP-dependent activation of transepithelial chloride secretion is at least partially preserved in cultured CF airway epithelium.

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. 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

    PubMed  CAS  Google Scholar 

  2. Bijman J, Quinton PM (1984) Influence of abnormal CL impermeability on sweating in cystic fibrosis. Am J Physiol 247: C3-C9

    PubMed  CAS  Google Scholar 

  3. Bijman J, Englert HC, Lang HJ, Fromter E (1987) Characterisation of human sweat duct chloride conductance by chloride channel blockers. Pflügers Arch 408:511–514

    Article  PubMed  CAS  Google Scholar 

  4. Boucher RC, Stutts MJ, Knowles MR, Cantley L, Gatzy JT (1986) Na+ transport in cystic fibrosis respiratory epithelia. Abnormal basal rate and response to adenylate cyclase activation. J Clin Invest 78:1245–1252

    PubMed  CAS  Google Scholar 

  5. Boucher RC, Willumsen NJ, Knowles MR, Yankaskas J, Gatzy JT (1988) Na and Cl absorption in respiratory epithelia: the role of apical and basolateral membranes. In: Gianni M, Quinton PM (eds) Cellular and molecular basis of cystic fibrosis. San Francisco Press, San Francisco, pp 107–114

    Google Scholar 

  6. Cotton CU, Stutts MJ, Knowles MR, Gatzy JT, Boucher RC (1987) Abnormal apical cell membrane in cystic fibrosis respiratory epithelium. An in vitro electrophysiologic analysis. J Clin Invest 79:80–85

    PubMed  CAS  Google Scholar 

  7. De Jonge HR, Bijman J, Sinaasappel M (1987) Relation of regulatory enzyme levels to chloride transport in intestinal epithelial cells. Pediatr Pulmonol [Suppl] 1:54–57

    Google Scholar 

  8. De Jonge HR, Bijman J, Sinaasappel M (1988) Application of regulatory principles to CF. Pediatr Pulmonol [Suppl] 2:92–95

    Google Scholar 

  9. Dharmsathaphorn K, Mandel KG, Masui H, McRoberts JA (1985) Vasoactive intestinal polypeptide-induced chloride secretion by a colonic epithelial cell line. J Clin Invest 75:462–471

    PubMed  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  12. Jongkind JF, Verkerk A, Visser WJ, Van Dongen JM (1982) Isolation of autofluorescent aged human fibroblasts by flow sorting. Exp Cell Res 138:409–417

    Article  PubMed  CAS  Google Scholar 

  13. Knowles MR, Stutts MJ, Spock A, Fischer N, Gatzy JT, Boucher RC (1983) Abnormal ion permeation through cystic fibrosis respiratory epithelium. Science 221:1067–1070

    Article  PubMed  CAS  Google Scholar 

  14. Knowles MR, Stutts MJ, Cotton CU, Earp HS, Yankaskas JR, Gatzy JT, Boucher RC (1988) Chloride secretion across normal and cystic fibrosis respiratory epithelia. In: Gianni M, Quinton PM (eds) Cellular and molecular basis of cystic fibrosis. San Francisco Press, San Francisco, pp 165–170

    Google Scholar 

  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  17. Liedtke CM, Romero M, Hopfer U (1988) Adrenergic control of the Na, K, 2Cl cotransporter in airway epithelial cells. In: Gianni M, Quinton PM (eds) Cellular and molecular basis of cystic fibrosis. San Francisco Press, San Francisco, pp 307–313

    Google Scholar 

  18. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  19. McPherson MA, Dormer RL (1987) The molecular and biochemical basis of cystic fibrosis. Biosci Rep 7:167–185

    Article  PubMed  CAS  Google Scholar 

  20. Moll R, Franke WW, Volc-Platzer B, Krepler R (1982) Different keratin polypeptides in epidermis and other epithelia and human skin. J Cell Biol 95:285–295

    Article  PubMed  CAS  Google Scholar 

  21. Rheinwald RH, Green H (1975) Serial cultivation of strains of human epidermal keratinocytes. Cell 6:331–344

    Article  PubMed  CAS  Google Scholar 

  22. Stutts MJ, Cotton CU, Yankaskas JR, Cheng E, Knowles MR, Gatzy JT, Boucher RC (1985) Chloride uptake into cultured airway epithelial cells from cystic fibrosis patients and normal individuals. Proc Natl Acad Sci USA 82:6677–6681

    Article  PubMed  CAS  Google Scholar 

  23. Taylor CJ, Baxter PS, Hardcastle J, Hardcastle PT (1987) Absence of secretory response in jejunal biopsy samples from children with cystic fibrosis. Lancet 1:107–108

    Article  Google Scholar 

  24. Towbin H, Staehelin T, Gordin J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Proc Natl Acad Sci USA 76:4350–4354

    Article  PubMed  CAS  Google Scholar 

  25. Van Muyen GNP, Ruiter DJ, Franke WW, Achtstaetter T, Haasnoot WHB, Ponec M, Warnaar SO (1986) Cell type heterogeneity of cytokeratin expression in complex epithelia and carcinomas as demonstrated by monoclonal antibodies specific for cytokeratins 4 and 13. Exp Cell Res 162:97–113

    Article  Google Scholar 

  26. Welsh MJ, Fick RB (1987) Cystic fibrosis (perspectives). J Clin Invest 80:1523–1526

    Article  PubMed  CAS  Google Scholar 

  27. Widdicombe JH (1986) Cystic fibrosis and beta-adrenergic response of airway epithelial cell cultures. Am J Physiol 251: R818-R222

    PubMed  CAS  Google Scholar 

  28. Willumsen NJ, Boucher RC (1989) Activation of an apical chloride conductance by Ca ionophores in CF airway epithelial cells. Am J Physiol 256:C226-C233

    PubMed  CAS  Google Scholar 

  29. Willumsen NJ, Boucher RC (1989) Shunt resistances and ion permeabilities in normal and cystic fibrosis airway epithelia. Am J Physiol 256:C1054-C1063

    PubMed  CAS  Google Scholar 

  30. Willumsen NJ, Davis W, Boucher RC (1989) Intracellular Clactivity and cellular Cl-pathways in cultured human airway epithelium. Am J Physiol 256:C1033-C1044

    PubMed  CAS  Google Scholar 

  31. Yankaskas JR, Cotton CU, Knowles MR, Gatzy JT, Boucher RC (1985) Culture of human nasal epithelial cells on collagen matrix supports. A comparison of bioelectric properties of normal and cystic fibrosis epithelia. Am Rev Respir Dis 132:1281–1287

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cell Biology and Genetics, Erasmus Universiteit, P/O box 1738, NL-3000, DR Rotterdam, The Netherlands

    Elly Verbeek, Jan Bijman, Joke Keulemans, Arthur W. M. van der Kamp & Bob J. Scholte

  2. Department of Biochemistry I, Erasmus Universiteit, P/O box 1738, NL-3000, DR Rotterdam, The Netherlands

    Hugo R. de Jonge

  3. Department of Pediatrics, Erasmus Universiteit, P/O box 1738, NL-3000, DR Rotterdam, The Netherlands

    Maarten Sinaasappel

Authors
  1. Elly Verbeek
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hugo R. de Jonge
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jan Bijman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Joke Keulemans
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Maarten Sinaasappel
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Arthur W. M. van der Kamp
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bob J. Scholte
    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

Verbeek, E., de Jonge, H.R., Bijman, J. et al. Chloride transport in cultured nasal epithelium of cystic fibrosis patients. Pflugers Arch. 415, 540–546 (1990). https://doi.org/10.1007/BF02583504

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02583504

Key words

Search

Navigation