1887

Abstract

Tight junctions, paracellular permeability barriers that define epithelial cell polarity, play an essential role in transepithelial transport, cell–cell adhesion and lymphocyte transmigration. They are also important for the maintenance of innate immune defence and intestinal antigen uptake. Ammonium () is elevated in the gastric aspirates of -infected patients and has been implicated in the disruption of tight-junction functional integrity and the induction of gastric mucosal damage during infection. The precise mechanism of the effect of ammonium and the molecular targets of ammonium in host tissue are not yet identified. To study the effects of ammonium on epithelial tight junctions, the human colon carcinoma cell line Caco-2 was cultured on permeable supports and the transepithelial resistance (TER) was measured at different time intervals following exposure to ammonium salts or -derived ammonium. A biphasic response to treatment with ammonium was found. Acute exposure to ammonium salts or NH/ derived from urea metabolism by wild-type resulted in a 20–30 % decrease in TER. After 24 h, the NHCl-treated cells showed a partial recovery of TER. In contrast, the control culture, or cultures that were exposed to supernatants derived from urease-deficient , showed no significant decrease in TER. Occludin-specific immunoblots revealed the expression of a low-molecular-weight form of occludin of 42 kDa upon NH/ exposure. The results indicate that modulation of tight-junction function by is ammonium-dependent and linked to the accumulation of a low-molecular-weight and detergent-soluble form of occludin.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.28049-0
2005-10-01
2019-10-22
Loading full text...

Full text loading...

/deliver/fulltext/micro/151/10/3267.html?itemId=/content/journal/micro/10.1099/mic.0.28049-0&mimeType=html&fmt=ahah

References

  1. Alexander, J. S., Dayton, T., Davis, C. & 9 other authors ( 1998; ). Activated T-lymphocytes express occludin, a component of tight junctions. Inflammation 22, 573–582.[CrossRef]
    [Google Scholar]
  2. Amieva, M. R., Vogelmann, R., Covacci, A., Tompkins, L. S., Nelson, W. J. & Falkow, S. ( 2003; ). Disruption of the epithelial apical-junctional complex by Helicobacter pylori CagA. Science 300, 1430–1434.[CrossRef]
    [Google Scholar]
  3. Blaser, M. J. & Atherton, J. C. ( 2004; ). Helicobacter pylori persistence: biology and disease. J Clin Invest 113, 321–333.[CrossRef]
    [Google Scholar]
  4. Bojarski, C., Weiske, J., Schöneberg, T. & 7 other authors ( 2004; ). The specific fates of tight junction proteins in apoptotic epithelial cells. J Cell Sci 117, 2097–2107.[CrossRef]
    [Google Scholar]
  5. Bruewer, M., Luegering, A., Kucharzik, T., Parkos, C. A., Madara, J. L., Hopkins, A. & Nusrat, A. ( 2003; ). Proinflammatory cytokines disrupt epithelial barrier function by apoptosis-independent mechanisms. J Immunol 171, 6164–6172.[CrossRef]
    [Google Scholar]
  6. Chong, S., Dando, S. A. & Morrison, R. A. ( 1997; ). Evaluation of Biocoat intestinal epithelium differentiation environment (3-day cultured Caco-2 cells) as an absorption screening model with improved productivity. Pharm Res 12, 1835–1837.
    [Google Scholar]
  7. Eaton, K. A., Brooks, C. L., Morgan, D. R. & Krakowka, S. ( 1991; ). Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect Immun 59, 2470–2475.
    [Google Scholar]
  8. Gebert, B., Fischer, W., Weiss, E., Hoffmann, R. & Haas, R. ( 2003; ). Helicobacter pylori vacuolating cytotoxin inhibits T lymphocyte activation. Science 301, 1099–1102.[CrossRef]
    [Google Scholar]
  9. Gillen, A. D., Wirz, A. A., Neithercut, W. D., Ardill, J. E. S. & McColl, K. E. L. ( 1999; ). Helicobacter pylori infection potentiates the inhibition of gastric acid secretion by omeprazole. Gut 44, 468–475.[CrossRef]
    [Google Scholar]
  10. Guillemin, K., Salama, N. R., Tompkins, L. S. & Falkow, S. ( 2002; ). Cag pathogenicity island-specific responses of gastric epithelial cells to Helicobacter pylori infection. Proc Natl Acad Sci U S A 99, 15136–15141.[CrossRef]
    [Google Scholar]
  11. Hagen, A. S., Wu, J. H. & Morrison, S. W. ( 2000; ). NH4Cl inhibition of acid secretion: possible involvement of an apical K+ channel in bullfrog oxyntic cells. Am J Physiol Gastrointest Liver Physiol 279, G400–G410.
    [Google Scholar]
  12. Handlogten, M. E., Hong, S.-P., Westhoff, C. M. & Weiner, D. ( 2004; ). Basolateral ammonium transport by the mouse inner medullary collecting duct cell (mIMCD-3). Am J Physiol Renal Physiol 287, F628–F638.[CrossRef]
    [Google Scholar]
  13. Ivanov, A. I., Nusrat, A. & Parkos, C. ( 2004; ). Endocytosis of epithelial apical junctional proteins by a clathrin-mediated pathway into a unique storage compartment. Mol Biol Cell 15, 176–188.
    [Google Scholar]
  14. Kirsten, E., Gerez, C. & Kirsten, R. ( 1963; ). An enzymatic microdetermination method for ammonia, specifically for extracts of animal tissues and fluids. Determination of NH4 ions in blood. Biochem Z 337, 312–319.
    [Google Scholar]
  15. Kleiner, D. ( 1981; ). The transport of NH3 and NH4 + across biological membranes. Biochim Biophys Acta 639, 41–52.[CrossRef]
    [Google Scholar]
  16. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  17. Li, N., Lewis, P., Samuelson, D., Liboni, K. & Neu, J. ( 2004; ). Glutamine regulates CACO-2 cell tight junction proteins. Am J Physiol Gastrointest Liver Physiol 287, G726–G733.[CrossRef]
    [Google Scholar]
  18. Lichtenberger, L. M. & Romero, J. J. ( 1994; ). Effect of ammonium ion on the hydrophobic and barrier properties of the gastric mucus gel layer: implications on the role of ammonium in H. pylori-induced gastritis. J Gastroenterol Hepatol 9, S13–S19.[CrossRef]
    [Google Scholar]
  19. Mankertz, J., Waller, J. S., Hillenbrand, B., Tavalali, S., Florian, P., Schöneberg, T., Fromm, M. & Schulzke, J. D. ( 2002; ). Gene expression of the tight junction protein occludin includes differential splicing and alternative promoter usage. Biochem Biophys Res Comm 298, 657–666.[CrossRef]
    [Google Scholar]
  20. Nusrat, A., Turner, J. R. & Madara, J. L. ( 2000a; ). Molecular physiology and pathophysiology of tight junctions IV. Regulation of tight junctions by extracellular stimuli: nutrients, cytokines, and immune cells. Am J Physiol Gastrointest Liver Physiol 279, G851–G857.
    [Google Scholar]
  21. Nusrat, A., Chen, J. A., Foley, C. S., Liang, T. W., Tom, J., Cromwell, M., Quan, C. & Mrsny, R. J. ( 2000b; ). The coiled-coil domain of occludin can act to organise structural and functional elements of the epithelial tight junction. J Biol Chem 275, 29816–29822.[CrossRef]
    [Google Scholar]
  22. Papini, E., Satin, B., Norais, N., de Bernard, M., Telford, J. L., Rappuoli, R. & Montecucco, C. ( 1998; ). Selective increase of the permeability of polarized epithelial cell monolayers by Helicobacter pylori vacuolating toxin. J Clin Invest 102, 813–820.[CrossRef]
    [Google Scholar]
  23. Rieder, G., Einsiedl, W., Hatz, R. A., Stolte, M., Enders, G. A. & Walz, A. ( 2001; ). Comparison of CXC chemokines ENA-78 and interleukin-8 expression in Helicobacter pylori-associated gastritis. Infect Immun 69, 81–88.[CrossRef]
    [Google Scholar]
  24. Sidebotham, R. L., Worku, M., Karim, Q., Najma, Q., Dhir, N., Baron, K. & Hugh, J. ( 2003; ). How Helicobacter pylori urease may affect external pH and influence growth and mobility in the mucus environment: evidence from in vitro studies. Eur J Gastroenterol Hepatol 15, 395–401.[CrossRef]
    [Google Scholar]
  25. Singh, U., Van Itallie, C. M., Mitie, L. L., Anderson, J. M. & McClane, B. A. ( 2000; ). CaCo-2 cells treated with Clostridium perfringens enterotoxin form multiple large complex species, one of which contains the tight junctional protein occludin. J Biol Chem 275, 18407–18417.[CrossRef]
    [Google Scholar]
  26. Smoot, D. T., Mobley, H. L. T., Chippendale, G. R., Lewison, J. F. & Resau, J. H. ( 1990; ). Helicobacter pylori urease activity is toxic to human gastric epithelial cells. Infect Immun 58, 1992–1994.
    [Google Scholar]
  27. Stingl, K., Altendorf, K. & Bakker, E. P. ( 2002; ). Acid survival of Helicobacter pylori: how does urease activity trigger cytoplasmic pH homeostasis? Trends Microbiol 10, 70–74.[CrossRef]
    [Google Scholar]
  28. Suzuki, H., Yanaka, A. & Muto, H. ( 2000; ). Luminal ammonia retards restitution of guinea pig injured gastric mucosa in vitro. Am J Physiol Gastrointest Liver Physiol 279, G107–G117.
    [Google Scholar]
  29. Suzuki, H., Yanaka, A., Shibahara, T., Matsui, H., Nakahara, A., Tanaka, N., Muto, H., Momoi, T. & Uchiyama, Y. ( 2002; ). Ammonia-induced apoptosis is accelerated at higher pH in gastric surface mucous cells. Am J Physiol Gastrointest Liver Physiol 283, 986–995.[CrossRef]
    [Google Scholar]
  30. Tavelin, S., Hashimoto, K., Malkinson, J., Lazorova, L., Toth, I. & Artursson, P. ( 2003; ). A new principle for tight junction modulation based on occludin peptides. Mol Pharmacol 64, 1530–1540.[CrossRef]
    [Google Scholar]
  31. Terres, A. M., Windle, H. J., Ardini, E. & Kelleher, D. P. ( 2003; ). Soluble extracts from Helicobacter pylori induce dome formation in polarized intestinal epithelial monolayers in a laminin-dependent manner. Infect Immun 71, 4067–4078.[CrossRef]
    [Google Scholar]
  32. Triebling, A. T., Korsten, M. A., Dlugosz, J. W., Paronetto, F. & Lieber, C. S. ( 1991; ). Severity of Helicobacter-induced gastric injury correlates with gastric juice ammonia. Dig Dis Sci 36, 1089–1096.[CrossRef]
    [Google Scholar]
  33. Vastag, M., Neuhofer, W., Nagel, W. & Beck, F. X. ( 2005; ). Ammonium affects tight junctions and the cytoskeleton in MDCK cells. Pflügers Archiv Eur J Physiol 449, 384–391.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.28049-0
Loading
/content/journal/micro/10.1099/mic.0.28049-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error