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Pflügers Archiv

, Volume 423, Issue 3–4, pp 213–220 | Cite as

Characterisation of volume-activated ion transport across epithelial monolayers of human intestinal T84 cells

  • G. T. A. McEwan
  • C. D. A. Brown
  • B. H. Hirst
  • N. L. Simmons
Transport Processes, Metabolism and Endocrinology; Kidney, Gastrointestinal Tract, and Exocrine Glands

Abstract

The effects of hypo-osmolarity upon transepithelial ion transport in human intestinal cell layers have been investigated. Exposure of the basal-lateral surfaces to hypo-osmotic media resulted in a transient stimulation of inward short-circuit current (Isc). This transient stimulation of inward current by hypo-osmotic media was abolished by 100 μmol/l 4,4′-diisothiocyanostilbene 2,2′-disulphonic acid (DIDS). After prestimulation of inward Isc by vasoactive intestinal peptide (VIP) or by combinations of carbachol and prostaglandin E1 hypoosmotic exposure of the basal-lateral surfaces resulted in a further transient stimulation of Isc. The stimulation of Isc in these conditions was largely insensitive to DIDS inhibition. Exposure of the basal-lateral surfaces to hypo-osmotic media resulted in a stimulation of loop-diuretic-insensitive 86Rb efflux across the basal-lateral surfaces. In addition, hypo-osmotic exposure of T84 cells is also associated with an increase in cytosolic Ca2+. It is concluded that the effects of hypo-osmotic exposure of T84 cells on secretory Isc are consistent with the activation of a DIDS-sensitive apical Cl conductance and a basal-lateral K+ conductance. With prior activation of inward Isc by VIP via a cAMP-activated DIDS-insensitive apical Cl conductance, augmentation of the secretory current by hypo-osmotic exposure is likely to result primarily from increased basal-lateral K+ current and loop-diuretic-sensitive Cl uptake.

Key words

Cell volume regulation T84 cell Intestinal epithelial cell Cl secretion 

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References

  1. 1.
    Aiton JF, Brown CDA, Ogden P, Simmons NL (1982) K+ transport in “tight” epithelial monolayers of MDCK cells. J Membr Biol 65:99–109Google Scholar
  2. 2.
    Andersen MP, Welsh MJ (1991) Calcium and cAMP activate different chloride channels in the apical membrane of normal and cystic fibrosis epithelia. Proc Natl Acad Sci USA 88:6003–6007Google Scholar
  3. 3.
    Butt AG, Clapp WL, Frizzell RA (1990) Potassium conductances in tracheal epithelium activated by secretion and cell swelling. Am J Physiol 258:C630-C638Google Scholar
  4. 4.
    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 76:1837–1842Google Scholar
  5. 5.
    Cliff WH, Frizzell RA (1990) Separate Cl conductances activated by cAMP and Ca2+ in Cl-secreting epithelial cells. Proc Natl Acad Sci USA 87:4956–4960Google Scholar
  6. 6.
    Dharmsathaphorn K, Pandol SJ (1986) Mechanism of chloride secretion induced by carbachol in a colonic epithelial cell line. J Clin Invest 77:348–354Google Scholar
  7. 7.
    Dharmsathaphorn K, McRoberts JA, Mandel KG, Tisdale LD, Masui H (1984) A human tumor cell line that maintains vectorial electrolyte transport. Am J Physiol 246:G204-G208Google Scholar
  8. 8.
    Dharmsathaphorn K, Mandel KG, Masui H, McRoberts JA (1985) Vasoactive intestinal polypeptide-induced chloride secretion by a colonic epithelial cell-line. Direct participation of a basolaterally localized Na+, K+, Cl cotransport system. J Clin Invest 75:462–471Google Scholar
  9. 9.
    Giraldez F, Valverde MA, Sepulveda FV (1988) Hypotonicity increases apical membrane Cl conductance in Necturus enterocytes. Biochim Biophys Acta 942:353–356Google Scholar
  10. 10.
    Haas M (1989) Properties and diversity of Na+/K+/Cl cotransporters. Annu Rev Physiol 51:443–457Google Scholar
  11. 11.
    Hazama A, Okada Y (1988) Ca2+ sensitivity of volume regulatory K+ and Cl channels in cultured human epithelial cells. J Physiol (Lond) 402:687–702Google Scholar
  12. 12.
    Hurst AM, Hunter M (1990) Stretch activated channels in single early distal tubule cells of the frog. J Physiol (Lond) 430:13–25Google Scholar
  13. 13.
    Kase H, Wakui M, Petersen OH (1991) Stimulatory and inhibitory actions of VIP and cyclic AMP on cytoplasmic Ca2+ signal generation in pancreatic acinar cells. Pflügers Arch 419:668–670Google Scholar
  14. 14.
    Kolb HA, Paulmichl M, Lang F (1987) Epinephrine activates an outward rectifying K+ channel in Madin Darby canine kidney cells. Pflügers Arch 408:584–591Google Scholar
  15. 15.
    Luckhoff A (1986) Measuring cytosolic free calcium concentration with indo-1: the pitfalls of using the ratio of two fluorescence intensities recorded at different wavelengths. Cell Calcium 7:233–248Google Scholar
  16. 16.
    McCann JD, Welsh MJ (1989) Identification and regulation of whole-cell chloride currents in airway epithelium. J Gen Physiol 94:1015–1036Google Scholar
  17. 17.
    McEwan GTA, Brown CDA, Hirst BH, Simmons NL (1992) Hypo-osmolar stimulation of transepithelial Cl secretion in cultured human T84 intestinal epithelial layers. Biochim Biophys Acta 1135:180–183Google Scholar
  18. 18.
    McEwan GTA, Hunter J, Hirst BH, Simmons NL (1992) Volume-activated Cl secretion and transepithelial vinblastine secretion mediated by P-glycoprotein are not correlated in cultured human T84 intestinal epithelial layers. FEBS Lett 304:233–236Google Scholar
  19. 19.
    Riordan JR, Rommens JM, Kerem B-S, Alon N, Rozmahel R, Grzelczak Z, Zielenski J, Lok S, Plavsic N, Chou J-L, Drumm MM, Iannuzzi MC, Collins FS, Tsui L-C (1989) Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science 241:1066–1072Google Scholar
  20. 20.
    Ritter M, Paulmichl M, Lang F (1991) Further characterisation of volume regulatory decrease in cultured renal epitheloid cells. Pflügers Arch 418:35–39Google Scholar
  21. 21.
    Simmons NL (1990) Tissue culture of established renal cell lines. Methods Enzymol 191:426–437Google Scholar
  22. 22.
    Simmons NL (1991) The effect of hypo-osmolarity upon transepithelial ion transport in cultured renal epithelial layers (MDCK). Pflügers Arch 419:572–578Google Scholar
  23. 23.
    Solc CK, Wine JJ (1991) Swelling-induced and depolarization-induced Cl channels in normal and cystic fibrosis epithelial cells. Am J Physiol 261:C658-C674Google Scholar
  24. 24.
    Sorscher EJ, Fuller CM, Bridges RJ, Tousson A, Marchase RB, Brinkley BR, Frizzell RA, Benos DJ (1992) Identification of a membrane protein from T84 cells using antibodies made against a DIDS-binding peptide. Am J Physiol 262:C136-C147Google Scholar
  25. 25.
    Tabcharani JA, Low W, Elie D, Hanrahan JW (1990) Low-conductance chloride channel activated by cAMP in the epithelial cell-line T84. FEBS Lett 270:157–164Google Scholar
  26. 26.
    Thiemann A, Grunder S, Pusch M, Jentsch TJ (1992) A chloride channel widely expressed in epithelial and non-epithelial cells. Nature 356:57–60Google Scholar
  27. 27.
    Welsh MJ (1986) Adrenergic regulation of ion transport by primary cultures of canine tracheal epithelium: Cellular electrophysiology. J Membr Biol 91:121–128Google Scholar
  28. 28.
    Weymer A, Huott P, Liu W, McRoberts JA, Dharmsathaphorn K (1985) Chloride secretory mechanism induced by prostaglandin E1 in a colonic epithelial cell-line. J Clin Invest 76:1828–1836Google Scholar
  29. 29.
    Widdicombe JH, Wine JJ (1991) The basic defect in cystic fibrosis. Trends Biochem Sci 16:474–477Google Scholar
  30. 30.
    Williams DA, Fay FS (1990) Intracellular calibration of the fluorescent calcium indicator Fura-2. Cell Calcium 11:75–83Google Scholar
  31. 31.
    Wong SME, Tesfaye A, DeBell MC, Chase HS (1990) Carbachol increases basolateral K+ conductance in T84 cells. Simultaneous measurements of cell Ca2+ and GK explore calcium's role. J Gen Physiol 96:1271–1285Google Scholar
  32. 32.
    Worrell RT, Butt GA, Cliff WH, Frizzell RA (1989) A volume-sensitive chloride conductance in human colonic cell line T84. Am J Physiol 256:C1111-C1119Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • G. T. A. McEwan
    • 1
  • C. D. A. Brown
    • 1
  • B. H. Hirst
    • 1
  • N. L. Simmons
    • 1
  1. 1.Gastrointestinal Drug Delivery Research Centre, Department of Physiological SciencesUniversity of Newcastle upon Tyne, The Medical SchoolNewcastle upon TyneUK

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