Abstract
Escherichia coli heat stable enterotoxin (STa) caused Cl- secretion across T84 cell monolayers in a dose-dependent manner only when applied to the apical membrane surface and not when applied to the basolateral surface. Measurement of cAMP, cGMP, and free cytosolic Ca2+ in response to STa suggested that cGMP alone mediated the Cl- secretory response. Studies utilizing blockers of the Na+,K+-ATPase pump, a Na+,K+,Cl- cotransport system, a K+ channel, and a Cl- channel suggest that all of them participate in the Cl- secretory process induced by STa. The results suggest that the Cl- secretory response induced by STa is mediated by cGMP after the enterotoxin binds to its receptor on the apical membrane. The enterotoxin, by increasing cGMP, opens a K+ channel on the basolateral membrane as well as a Cl- channel on the apical membrane. The activation of these ion exit mechanisms, together with activations of the Na+,K+,Cl- cotransporter and the Na+,K+-ATPase pump drives Cl- exit through the Cl- channel on the apical membrane.
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Cartwright C. A., McRoberts J. A., Mandel K. G., Dharmsathaphorn K. Synergistic action of cyclic adenosine monophosphate- and calcium-mediated chloride secretion in a colonic epithelial cell line. J Clin Invest. 1985 Nov;76(5):1837–1842. doi: 10.1172/JCI112176. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dharmsathaphorn K., Huott P., Cartwright C. A., McRoberts J. A., Mandel K. G., Beuerlein G. Inhibition of ATP levels by quinidine in a human colonic epithelial cell line. Am J Physiol. 1986 Jun;250(6 Pt 1):G806–G813. doi: 10.1152/ajpgi.1986.250.6.G806. [DOI] [PubMed] [Google Scholar]
- Dharmsathaphorn K., Mandel K. G., Masui H., McRoberts J. A. 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. 1985 Feb;75(2):462–471. doi: 10.1172/JCI111721. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dharmsathaphorn K., Pandol S. J. Mechanism of chloride secretion induced by carbachol in a colonic epithelial cell line. J Clin Invest. 1986 Feb;77(2):348–354. doi: 10.1172/JCI112311. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Garty H., Rudy B., Karlish S. J. A simple and sensitive procedure for measuring isotope fluxes through ion-specific channels in heterogenous populations of membrane vesicles. J Biol Chem. 1983 Nov 10;258(21):13094–13099. [PubMed] [Google Scholar]
- Giannella R. A., Drake K. W. Effect of purified Escherichia coli heat-stable enterotoxin on intestinal cyclic nucleotide metabolism and fluid secretion. Infect Immun. 1979 Apr;24(1):19–23. doi: 10.1128/iai.24.1.19-23.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grinstein S., Rothstein A., Sarkadi B., Gelfand E. W. Responses of lymphocytes to anisotonic media: volume-regulating behavior. Am J Physiol. 1984 Mar;246(3 Pt 1):C204–C215. doi: 10.1152/ajpcell.1984.246.3.C204. [DOI] [PubMed] [Google Scholar]
- Guandalini S., Migliavacca M., de Campora E., Rubino A. Cyclic guanosine monophosphate effects on nutrient and electrolyte transport in rabbit ileum. Gastroenterology. 1982 Jul;83(1 Pt 1):15–21. [PubMed] [Google Scholar]
- Guarino A., Cohen M., Thompson M., Dharmsathaphorn K., Giannella R. T84 cell receptor binding and guanyl cyclase activation by Escherichia coli heat-stable toxin. Am J Physiol. 1987 Dec;253(6 Pt 1):G775–G780. doi: 10.1152/ajpgi.1987.253.6.G775. [DOI] [PubMed] [Google Scholar]
- Madara J. L., Dharmsathaphorn K. Occluding junction structure-function relationships in a cultured epithelial monolayer. J Cell Biol. 1985 Dec;101(6):2124–2133. doi: 10.1083/jcb.101.6.2124. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Mandel K. G., McRoberts J. A., Beuerlein G., Foster E. S., Dharmsathaphorn K. Ba2+ inhibition of VIP- and A23187-stimulated Cl- secretion by T84 cell monolayers. Am J Physiol. 1986 Mar;250(3 Pt 1):C486–C494. doi: 10.1152/ajpcell.1986.250.3.C486. [DOI] [PubMed] [Google Scholar]
- 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]
- O'Grady S. M., Field M., Nash N. T., Rao M. C. Atrial natriuretic factor inhibits Na-K-Cl cotransport in teleost intestine. Am J Physiol. 1985 Nov;249(5 Pt 1):C531–C534. doi: 10.1152/ajpcell.1985.249.5.C531. [DOI] [PubMed] [Google Scholar]
- Poenie M., Alderton J., Tsien R. Y., Steinhardt R. A. Changes of free calcium levels with stages of the cell division cycle. Nature. 1985 May 9;315(6015):147–149. doi: 10.1038/315147a0. [DOI] [PubMed] [Google Scholar]
- Rangachari P. K. Ba++ on the resting frog stomach: effects on electrical and secretory parameters. Am J Physiol. 1975 Feb;228(2):511–517. doi: 10.1152/ajplegacy.1975.228.2.511. [DOI] [PubMed] [Google Scholar]
- Rao M. C., Nash N. T., Field M. Differing effects of cGMP and cAMP on ion transport across flounder intestine. Am J Physiol. 1984 Jan;246(1 Pt 1):C167–C171. doi: 10.1152/ajpcell.1984.246.1.C167. [DOI] [PubMed] [Google Scholar]
- Rao M. C., Orellana S. A., Field M., Robertson D. C., Giannella R. A. Comparison of the biological actions of three purified heat-stable enterotoxins: effects on ion transport and guanylate cyclase activity in rabbit ileum in vitro. Infect Immun. 1981 Jul;33(1):165–170. doi: 10.1128/iai.33.1.165-170.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spiegel A. M., Gierschik P., Levine M. A., Downs R. W., Jr Clinical implications of guanine nucleotide-binding proteins as receptor-effector couplers. N Engl J Med. 1985 Jan 3;312(1):26–33. doi: 10.1056/NEJM198501033120106. [DOI] [PubMed] [Google Scholar]
- Staples S. J., Asher S. E., Giannella R. A. Purification and characterization of heat-stable enterotoxin produced by a strain of E. coli pathogenic for man. J Biol Chem. 1980 May 25;255(10):4716–4721. [PubMed] [Google Scholar]
- Thompson M. R., Giannella R. A. Revised amino acid sequence for a heat-stable enterotoxin produced by an Escherichia coli strain (18D) that is pathogenic for humans. Infect Immun. 1985 Mar;47(3):834–836. doi: 10.1128/iai.47.3.834-836.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson M. R., Luttrell M., Overmann G., Giannella R. A. Biological and immunological characteristics of 125I-4Tyr and -18Tyr Escherichia coli heat-stable enterotoxin species purified by high-performance liquid chromatography. Anal Biochem. 1985 Jul;148(1):26–36. doi: 10.1016/0003-2697(85)90623-2. [DOI] [PubMed] [Google Scholar]
- Tsien R. Y., Rink T. J., Poenie M. Measurement of cytosolic free Ca2+ in individual small cells using fluorescence microscopy with dual excitation wavelengths. Cell Calcium. 1985 Apr;6(1-2):145–157. doi: 10.1016/0143-4160(85)90041-7. [DOI] [PubMed] [Google Scholar]
- Wasserman S. I., Barrett K. E., Huott P. A., Beuerlein G., Kagnoff M. F., Dharmsathaphorn K. Immune-related intestinal Cl- secretion. I. Effect of histamine on the T84 cell line. Am J Physiol. 1988 Jan;254(1 Pt 1):C53–C62. doi: 10.1152/ajpcell.1988.254.1.C53. [DOI] [PubMed] [Google Scholar]
- Weymer A., Huott P., Liu W., McRoberts J. A., Dharmsathaphorn K. Chloride secretory mechanism induced by prostaglandin E1 in a colonic epithelial cell line. J Clin Invest. 1985 Nov;76(5):1828–1836. doi: 10.1172/JCI112175. [DOI] [PMC free article] [PubMed] [Google Scholar]