Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Jul 15;88(14):6003–6007. doi: 10.1073/pnas.88.14.6003

Calcium and cAMP activate different chloride channels in the apical membrane of normal and cystic fibrosis epithelia.

M P Anderson 1, M J Welsh 1
PMCID: PMC52010  PMID: 1712478

Abstract

The genetic disease cystic fibrosis (CF) causes decreased Cl- transport in several epithelia. cAMP-dependent regulation of apical membrane Cl- channels is defective in CF airway epithelia; as a result, CF epithelia fail to secrete Cl-. In contrast, Ca(2+)-stimulated Cl- secretion is intact in CF airway epithelia and thus has the potential to bypass the CF Cl- secretory defect. For a Cl- channel to govern Cl- secretion, it must be located in the apical membrane. To specifically investigate apical membrane Cl- channels, we studied cells grown on permeable filter supports and measured Cl- currents across the apical membrane. We found that Ca2+ and cAMP activate different Cl- channels in the apical membrane. (i) Ca(2+)-activated Cl- channels were present in the apical membrane of airway but not in intestinal epithelia. (ii) cAMP- but not Ca(2+)-activated Cl- channels were defective in CF airway epithelia. (iii) Ca(2+)- but not cAMP-activated Cl- channels were blocked by 4,4'-diisothiocyanato-2,2'-stilbenedisulfonate. (iv) Ca(2+)- and cAMP-activated apical channels had different anion permeabilities. (v) An increase in both second messengers produced an additive increase in Cl- current. These results also explain the puzzling observation that Ca(2+)-stimulated Cl- secretion is defective in CF intestine: the Ca(2+)-activated Cl- channels that could circumvent the Cl- secretory defect in CF airway are missing from the apical membrane of intestinal epithelia.

Full text

PDF
6003

Images in this article

6006Image
on p.6006
6006Image
on p.6006
6007Image
on p.6007

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Augeron C., Laboisse C. L. Emergence of permanently differentiated cell clones in a human colonic cancer cell line in culture after treatment with sodium butyrate. Cancer Res. 1984 Sep;44(9):3961–3969. [PubMed] [Google Scholar]
  2. Berschneider H. M., Knowles M. R., Azizkhan R. G., Boucher R. C., Tobey N. A., Orlando R. C., Powell D. W. Altered intestinal chloride transport in cystic fibrosis. FASEB J. 1988 Jul;2(10):2625–2629. doi: 10.1096/fasebj.2.10.2838365. [DOI] [PubMed] [Google Scholar]
  3. Cliff W. H., Frizzell R. A. Separate Cl- conductances activated by cAMP and Ca2+ in Cl(-)-secreting epithelial cells. Proc Natl Acad Sci U S A. 1990 Jul;87(13):4956–4960. doi: 10.1073/pnas.87.13.4956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cotton C. U., Stutts M. J., Knowles M. R., Gatzy J. T., Boucher R. C. Abnormal apical cell membrane in cystic fibrosis respiratory epithelium. An in vitro electrophysiologic analysis. J Clin Invest. 1987 Jan;79(1):80–85. doi: 10.1172/JCI112812. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dawson D. C., Van Driessche W., Helman S. I. Osmotically induced basolateral K+ conductance in turtle colon: lidocaine-induced K+ channel noise. Am J Physiol. 1988 Jan;254(1 Pt 1):C165–C174. doi: 10.1152/ajpcell.1988.254.1.C165. [DOI] [PubMed] [Google Scholar]
  6. Devor D. C., Simasko S. M., Duffey M. E. Carbachol induces oscillations of membrane potassium conductance in a colonic cell line, T84. Am J Physiol. 1990 Feb;258(2 Pt 1):C318–C326. doi: 10.1152/ajpcell.1990.258.2.C318. [DOI] [PubMed] [Google Scholar]
  7. Grasset E., Bernabeu J., Pinto M. Epithelial properties of human colonic carcinoma cell line Caco-2: effect of secretagogues. Am J Physiol. 1985 May;248(5 Pt 1):C410–C418. doi: 10.1152/ajpcell.1985.248.5.C410. [DOI] [PubMed] [Google Scholar]
  8. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  9. Horn R., Marty A. Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol. 1988 Aug;92(2):145–159. doi: 10.1085/jgp.92.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jefferson D. M., Valentich J. D., Marini F. C., Grubman S. A., Iannuzzi M. C., Dorkin H. L., Li M., Klinger K. W., Welsh M. J. Expression of normal and cystic fibrosis phenotypes by continuous airway epithelial cell lines. Am J Physiol. 1990 Dec;259(6 Pt 1):L496–L505. doi: 10.1152/ajplung.1990.259.6.L496. [DOI] [PubMed] [Google Scholar]
  11. Madara J. L., Stafford J., Dharmsathaphorn K., Carlson S. Structural analysis of a human intestinal epithelial cell line. Gastroenterology. 1987 May;92(5 Pt 1):1133–1145. doi: 10.1016/s0016-5085(87)91069-9. [DOI] [PubMed] [Google Scholar]
  12. Mandel K. G., Dharmsathaphorn K., McRoberts J. A. Characterization of a cyclic AMP-activated Cl-transport pathway in the apical membrane of a human colonic epithelial cell line. J Biol Chem. 1986 Jan 15;261(2):704–712. [PubMed] [Google Scholar]
  13. McCann J. D., Bhalla R. C., Welsh M. J. Release of intracellular calcium by two different second messengers in airway epithelium. Am J Physiol. 1989 Aug;257(2 Pt 1):L116–L124. doi: 10.1152/ajplung.1989.257.2.L116. [DOI] [PubMed] [Google Scholar]
  14. McCann J. D., Li M., Welsh M. J. Identification and regulation of whole-cell chloride currents in airway epithelium. J Gen Physiol. 1989 Dec;94(6):1015–1036. doi: 10.1085/jgp.94.6.1015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McCann J. D., Matsuda J., Garcia M., Kaczorowski G., Welsh M. J. Basolateral K+ channels in airway epithelia. I. Regulation by Ca2+ and block by charybdotoxin. Am J Physiol. 1990 Jun;258(6 Pt 1):L334–L342. doi: 10.1152/ajplung.1990.258.6.L334. [DOI] [PubMed] [Google Scholar]
  16. McCann J. D., Welsh M. J. Basolateral K+ channels in airway epithelia. II. Role in Cl- secretion and evidence for two types of K+ channel. Am J Physiol. 1990 Jun;258(6 Pt 1):L343–L348. doi: 10.1152/ajplung.1990.258.6.L343. [DOI] [PubMed] [Google Scholar]
  17. McRoberts J. A., Beuerlein G., Dharmsathaphorn K. Cyclic AMP and Ca2+-activated K+ transport in a human colonic epithelial cell line. J Biol Chem. 1985 Nov 15;260(26):14163–14172. [PubMed] [Google Scholar]
  18. Quinton P. M. Cystic fibrosis: a disease in electrolyte transport. FASEB J. 1990 Jul;4(10):2709–2717. doi: 10.1096/fasebj.4.10.2197151. [DOI] [PubMed] [Google Scholar]
  19. Reinlib L., Mikkelsen R., Zahniser D., Dharmsathaphorn K., Donowitz M. Carbachol-induced cytosolic free Ca2+ increases in T84 colonic cells seen by microfluorimetry. Am J Physiol. 1989 Dec;257(6 Pt 1):G950–G960. doi: 10.1152/ajpgi.1989.257.6.G950. [DOI] [PubMed] [Google Scholar]
  20. Rich D. P., Anderson M. P., Gregory R. J., Cheng S. H., Paul S., Jefferson D. M., McCann J. D., Klinger K. W., Smith A. E., Welsh M. J. Expression of cystic fibrosis transmembrane conductance regulator corrects defective chloride channel regulation in cystic fibrosis airway epithelial cells. Nature. 1990 Sep 27;347(6291):358–363. doi: 10.1038/347358a0. [DOI] [PubMed] [Google Scholar]
  21. Riordan J. R., Rommens J. M., Kerem B., Alon N., Rozmahel R., Grzelczak Z., Zielenski J., Lok S., Plavsic N., Chou J. L. Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA. Science. 1989 Sep 8;245(4922):1066–1073. doi: 10.1126/science.2475911. [DOI] [PubMed] [Google Scholar]
  22. Smith J. J., McCann J. D., Welsh M. J. Bradykinin stimulates airway epithelial Cl- secretion via two second messenger pathways. Am J Physiol. 1990 Jun;258(6 Pt 1):L369–L377. doi: 10.1152/ajplung.1990.258.6.L369. [DOI] [PubMed] [Google Scholar]
  23. Smith P. L., Welsh M. J., Stoff J. S., Frizzell R. A. Chloride secretion by canine tracheal epithelium: I. Role of intracellular c AMP levels. J Membr Biol. 1982;70(3):217–226. doi: 10.1007/BF01870564. [DOI] [PubMed] [Google Scholar]
  24. Taylor C. J., Baxter P. S., Hardcastle J., Hardcastle P. T. Failure to induce secretion in jejunal biopsies from children with cystic fibrosis. Gut. 1988 Jul;29(7):957–962. doi: 10.1136/gut.29.7.957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Welsh M. J. Abnormal regulation of ion channels in cystic fibrosis epithelia. FASEB J. 1990 Jul;4(10):2718–2725. doi: 10.1096/fasebj.4.10.1695593. [DOI] [PubMed] [Google Scholar]
  26. Welsh M. J. Electrolyte transport by airway epithelia. Physiol Rev. 1987 Oct;67(4):1143–1184. doi: 10.1152/physrev.1987.67.4.1143. [DOI] [PubMed] [Google Scholar]
  27. Welsh M. J. Ion transport by primary cultures of canine tracheal epithelium: methodology, morphology, and electrophysiology. J Membr Biol. 1985;88(2):149–163. doi: 10.1007/BF01868429. [DOI] [PubMed] [Google Scholar]
  28. Widdicombe J. H. Cystic fibrosis and beta-adrenergic response of airway epithelial cell cultures. Am J Physiol. 1986 Oct;251(4 Pt 2):R818–R822. doi: 10.1152/ajpregu.1986.251.4.R818. [DOI] [PubMed] [Google Scholar]
  29. Widdicombe J. H., Welsh M. J., Finkbeiner W. E. Cystic fibrosis decreases the apical membrane chloride permeability of monolayers cultured from cells of tracheal epithelium. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6167–6171. doi: 10.1073/pnas.82.18.6167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Willumsen N. J., Boucher R. C. Activation of an apical Cl- conductance by Ca2+ ionophores in cystic fibrosis airway epithelia. Am J Physiol. 1989 Feb;256(2 Pt 1):C226–C233. doi: 10.1152/ajpcell.1989.256.2.C226. [DOI] [PubMed] [Google Scholar]
  31. de Jonge H. R., van den Berghe N., Tilly B. C., Kansen M., Bijman J. (Dys)regulation of epithelial chloride channels. Biochem Soc Trans. 1989 Oct;17(5):816–818. doi: 10.1042/bst0170816. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES