Pflügers Archiv

, Volume 451, Issue 3, pp 464–473 | Cite as

Water permeability in human airway epithelium

  • Peter Steen Pedersen
  • Kristina Procida
  • Per Leganger Larsen
  • Niels-Henrik Holstein-Rathlou
  • Ole Frederiksen
Ion Channels, Transporters

Abstract

Osmotic water permeability (Pf) was studied in spheroid-shaped human airway epithelia explants derived from nasal polyps by the use of a new improved tissue collection and isolation procedure. The fluid-filled spheroids were lined with a single cell layer with the ciliated apical cell membrane facing the outside. They were capable of surviving hours of experiment involving continuous superfusion of the bathing medium and changes of osmolarity. A new image analysis technique was developed for measuring the spheroid diameters, giving high time and measurement resolutions. The transepithelial Pf, determined by the changes of the apical solution osmolarity, was not influenced by the presence of glucose, Na+, or Na+/glucose-cotransport inhibitors in the bath, but was sensitive to the aquaporin (AQP) inhibitor HgCl2. The measured Pf levels and the values of activation energy were in the range of those seen in AQP-associated water transport. Together, these results indicate the presence of an AQP in the apical membrane of the spheroids. Notably, identical values for Pf were found in CF and non-CF airway preparations, as was the case also for the calculated spontaneous fluid absorption rates.

Keywords

Human airway epithelia Spheroids Cystic fibrosis Water permeability Fluid absorption Aquaporin 

References

  1. 1.
    Barry PH, Diamond JM (1984) Effects of unstirred layers on membrane phenomena. Physiol Rev 64:763–872PubMedGoogle Scholar
  2. 2.
    Boucher RC (1994) Human airway ion transport. Part one. Am J Respir Crit Care Med 150:271–281Google Scholar
  3. 3.
    Boucher RC (1994) Human airway ion transport. Part two. Am J Respir Crit Care Med 150:581–593Google Scholar
  4. 4.
    Boucher RC (2003) Regulation of airway surface liquid volume by human airway epithelia. Pflugers Arch 445:495–498PubMedGoogle Scholar
  5. 5.
    Carter EP, Umenishi F, Matthay MA, Verkman AS (1997) Developmental changes in water permeability across the alveolar barrier in perinatal rabbit lung. J Clin Invest 100:1071–1078PubMedGoogle Scholar
  6. 6.
    Castillon N, Hinnrasky J, Zahm JM, Kaplan H, Bonnet N, Corlieu P, Klossek JM, Taouil K, Avril-Delplanque A, Peault B, Puchelle E (2002) Polarized expression of cystic fibrosis transmembrane conductance regulator and associated epithelial proteins during the regeneration of human airway surface epithelium in three-dimensional culture. Lab Invest 82:989–998PubMedGoogle Scholar
  7. 7.
    Crews A, Taylor AE, Ballard ST (2001) Liquid transport properties of porcine tracheal epithelium. J Appl Physiol 91:797–802PubMedGoogle Scholar
  8. 8.
    Durand J, Durand-Arczynska W, Haab P (1981) Volume flow, hydraulic conductivity and electrical properties across bovine tracheal epithelium in vitro: effect of histamine. Pflugers Arch 392:40–45CrossRefPubMedGoogle Scholar
  9. 9.
    Farinas J, Kneen M, Moore M, Verkman AS (1997) Plasma membrane water permeability of cultured cells and epithelia measured by light microscopy with spatial filtering. J Gen Physiol 110:283–296CrossRefPubMedGoogle Scholar
  10. 10.
    Fischbarg J, Kuang KY, Vera JC, Arant S, Silverstein SC, Loike J, Rosen OM (1990) Glucose transporters serve as water channels. Proc Natl Acad Sci U S A 87:3244–3247PubMedGoogle Scholar
  11. 11.
    Folkesson HG, Matthay MA, Frigeri A, Verkman AS (1996) Transepithelial water permeability in microperfused distal airways. Evidence for channel-mediated water transport. J Clin Invest 97:664–671PubMedGoogle Scholar
  12. 12.
    Graham A, Steel DM, Alton EW, Geddes DM (1992) Second-messenger regulation of sodium transport in mammalian airway epithelia. J Physiol 453:475–491PubMedGoogle Scholar
  13. 13.
    Jayaraman S, Song Y, Verkman AS (2001) Airway surface liquid osmolality measured using fluorophore encapsulated liposomes. J Gen Physiol 117:423–430CrossRefPubMedGoogle Scholar
  14. 14.
    Jayaraman S, Song Y, Vetrivel L, Shankar L, Verkman AS (2001) Noninvasive in vivo fluorescence measurement of airway-surface liquid depth, salt concentration, and pH. J Clin Invest 107:317–324PubMedGoogle Scholar
  15. 15.
    Joris L, Quinton PM (1989) Evidence for electrogenic Na–glucose cotransport in tracheal epithelium. Pflugers Arch 415:118–120CrossRefPubMedGoogle Scholar
  16. 16.
    Joris L, Dab I, Quinton PM (1993) Elemental composition of human airway surface fluid in healthy and diseased airways. Am Rev Respir Dis 148:1633–1637PubMedGoogle Scholar
  17. 17.
    Jorissen M, Bessems A (1995) Normal ciliary beat frequency after ciliogenesis in nasal epithelial cells cultured sequentially as monolayer and in suspension. Acta Otolaryngol 115:66–70PubMedGoogle Scholar
  18. 18.
    Jun ES, Kim YS, Yoo, Roh HJ, Jung JS (2001) Changes in expression of ion channels and aquaporins mRNA during differentiation in normal human nasal epithelial cells. Life Sci 68:827–840CrossRefPubMedGoogle Scholar
  19. 19.
    King LS, Kozono D, Agre P (2004) From structure to disease: the evolving tale of aquaporin biology. Nat Rev Mol Cell Biol 5:687–698CrossRefPubMedGoogle Scholar
  20. 20.
    Krane CM, Fortner CN, Hand AR, McGraw DW, Lorenz JN, Wert SE, Towne JE, Paul RJ, Whitsett JA, Menon AG (2001) Aquaporin 5-deficient mouse lungs are hyperresponsive to cholinergic stimulation. Proc Natl Acad Sci U S A 98:14114–14119CrossRefPubMedGoogle Scholar
  21. 21.
    Kreda SM, Gynn MC, Fenstermacher DA, Boucher RC, Gabriel SE (2001) Expression and localization of epithelial aquaporins in the adult human lung. Am J Respir Cell Mol Biol 24:224–234PubMedGoogle Scholar
  22. 22.
    Landry JS, Eidelman DH (2001) Airway surface liquid: end of the controversy? J Gen Physiol 117:419–422CrossRefPubMedGoogle Scholar
  23. 23.
    Lee MD, King LS, Agre P (1997) The aquaporin family of water channel proteins in clinical medicine. Medicine (Baltimore) 76:141–156CrossRefGoogle Scholar
  24. 24.
    Marsh DJ, Jensen PK, Spring KR (1985) Computer-based determination of size and shape in living cells. J Microsc 137:281–292PubMedGoogle Scholar
  25. 25.
    Matsui H, Grubb BR, Tarran R, Randell SH, Gatzy JT, Davis CW, Boucher RC (1998) Evidence for periciliary liquid layer depletion, not abnormal ion composition, in the pathogenesis of cystic fibrosis airways disease. Cell 95:1005–1015CrossRefPubMedGoogle Scholar
  26. 26.
    Matsui H, Davis CW, Tarran R, Boucher RC (2000) Osmotic water permeabilities of cultured, well-differentiated normal and cystic fibrosis airway epithelia. J Clin Invest 105:1419–1427PubMedGoogle Scholar
  27. 27.
    Nielsen S, King LS, Christensen BM, Agre P (1997) Aquaporins in complex tissues. II. Subcellular distribution in respiratory and glandular tissues of rat. Am J Physiol 273:C1549–C1561PubMedGoogle Scholar
  28. 28.
    Novotny JA, Jakobsson E (1996) Computational studies of ion–water flux coupling in the airway epithelium. II. Role of specific transport mechanisms. Am J Physiol 270:C1764–C1772PubMedGoogle Scholar
  29. 29.
    Novotny JA, Jakobsson E (1996) Computational studies of ion–water flux coupling in the airway epithelium. I. Construction of model. Am J Physiol 270:C1751–C1763PubMedGoogle Scholar
  30. 30.
    Pedersen PS, Frederiksen O, Holstein-Rathlou NH, Larsen PL, Qvortrup K (1999) Ion transport in epithelial spheroids derived from human airway cells. Am J Physiol 276:L75–L80PubMedGoogle Scholar
  31. 31.
    Pedersen PS, Holstein-Rathlou NH, Larsen PL, Qvortrup K, Frederiksen O (1999) Fluid absorption related to ion transport in human airway epithelial spheroids. Am J Physiol 277:L1096–L1103PubMedGoogle Scholar
  32. 32.
    Saumon G, Seigne E, Clerici C (1990) Evidence for a sodium-dependent sugar transport in rat tracheal epithelium. Biochim Biophys Acta 1023:313–318PubMedGoogle Scholar
  33. 33.
    Smith JJ, Travis SM, Greenberg EP, Welsh MJ (1996) Cystic fibrosis airway epithelia fail to kill bacteria because of abnormal airway surface fluid. Cell 85:229–236CrossRefPubMedGoogle Scholar
  34. 34.
    Verkman AS (2000) Water permeability measurement in living cells and complex tissues. J Membr Biol 173:73–87CrossRefPubMedGoogle Scholar
  35. 35.
    Verkman AS, Matthay MA, Song Y (2000) Aquaporin water channels and lung physiology. Am J Physiol Lung Cell Mol Physiol 278:L867–L879PubMedGoogle Scholar
  36. 36.
    Welsh MJ (1987) Electrolyte transport by airway epithelia. Physiol Rev 67:1143–1184PubMedGoogle Scholar
  37. 37.
    Widdicombe JH, Bastacky SJ, Wu DX, Lee CY (1997) Regulation of depth and composition of airway surface liquid. Eur Respir J 10:2892–2897CrossRefPubMedGoogle Scholar
  38. 38.
    Willumsen NJ, Davis CW, Boucher RC (1994) Selective response of human airway epithelia to luminal but not serosal solution hypertonicity. Possible role for proximal airway epithelia as an osmolality transducer. J Clin Invest 94:779–787Google Scholar
  39. 39.
    Wu DX, Lee CY, Uyekubo SN, Choi HK, Bastacky SJ, Widdicombe JH (1998) Regulation of the depth of surface liquid in bovine trachea. Am J Physiol 274:L388–L395PubMedGoogle Scholar
  40. 40.
    Zabner J, Smith JJ, Karp PH, Widdicombe JH, Welsh MJ (1998) Loss of CFTR chloride channels alters salt absorption by cystic fibrosis airway epithelia in vitro. Mol Cell 2:397–403CrossRefPubMedGoogle Scholar
  41. 41.
    Zahm JM, Baconnais S, Davidson DJ, Webb S, Dorin J, Bonnet N, Balossier G, Puchelle E (2001) X-ray microanalysis of airway surface liquid collected in cystic fibrosis mice. Am J Physiol Lung Cell Mol Physiol 281:L309–L313PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Peter Steen Pedersen
    • 1
    • 2
  • Kristina Procida
    • 2
  • Per Leganger Larsen
    • 3
  • Niels-Henrik Holstein-Rathlou
    • 2
  • Ole Frederiksen
    • 2
  1. 1.Department of Clinical Genetics, RigshospitaletUniversity of CopenhagenCopenhagenDenmark
  2. 2.Department of Medical PhysiologyThe Panum Institute, University of CopenhagenCopenhagenDenmark
  3. 3.Department of Otolaryngology, Head and Neck Surgery, RigshospitaletUniversity of CopenhagenCopenhagenDenmark

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