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Epidermal Growth Factor Enhances Intracellular pH Regulation via Calcium Signaling in Acid-Exposed Primary Cultured Rabbit Gastric Epithelial Cells

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Abstract

We have elucidated the role of different ion transporters and epidermal growth factor(EGF) during luminal acid exposure in primary cultured rabbit surface epithelial cells by measuring intracellular calcium and pH. Amiloride, DIDS, or sodium or bicarbonate substitutions were used to inhibit ion transport. During luminal acid exposure the dominant intracellular pH regulator is the Na+/H+ antiport, and bicarbonate transport has only a secondary role, which is uncovered as the Na+/H+ function fails. The decrease in intracellular pH caused by luminal acid was significantly smaller in serosal EGF-treated epithelia than in controls. This defensive function of EGF was abolished by verapamil, BAPTA, and calmidazolium but not by TMB-8. EGF increased intracellular calcium, which was prevented by verapamil but not by TMB-8. EGF enhances gastric epithelial defense against luminal acid by inducing intracellular calcium signaling via plasma membrane verapamil-sensitive calcium channels and thereby enhancing the function of the Na+/H+ antiport.

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References

  1. Rossmann H, Sonnentag T, Heinzmann A, Seidler B, Bachmann O, Vieillard-Baron D, Gregor M, Seidler U (2001) Differential expression and regulation of Na(+)/H(+) exchanger isoforms in rabbit parietal and mucous cells. Am J Physiol Gastrointest Liver Physiol 281:G447–G458

    PubMed  CAS  Google Scholar 

  2. Rossmann H, Bachmann O, Vieillard-Baron D, Gregor M, Seidler U (1999) Na+/HCO3 − cotransport and expression of NBC1 and NBC2 in rabbit gastric parietal and mucous cells. Gastroenterology 116:1389–1398

    Article  PubMed  CAS  Google Scholar 

  3. Xu J, Barone S, Petrovic S, Wang Z, Seidler U, Riederer B, Ramaswamy K, Dudeja PK, Shull GE, Soleimani M (2003) Identification of an apical Cl−/HCO3 − exchanger in gastric surface mucous and duodenal villus cells. Am J Physiol Gastrointest Liver Physiol 285:G1225–G1234

    PubMed  CAS  Google Scholar 

  4. Rossmann H, Bachmann O, Wang Z, Shull GE, Obermaier B, Stuart-Tilley A, Alper SL, Seidler U (2001) Differential expression and regulation of AE2 anion exchanger subtypes in rabbit parietal and mucous cells. J Physiol 534:837–848

    Article  PubMed  CAS  Google Scholar 

  5. Puceat M (1999) pHi regulatory ion transporters: an update on structure, regulation and cell function. Cell Mol Life Sci 55:1216–1229

    Article  PubMed  CAS  Google Scholar 

  6. Orlowski J, Grinstein S (2004) Diversity of the mammalian sodium/proton exchanger SLC9 gene family [Review]. Pflugers Arch Eur J Physiol 447:549–565

    Article  CAS  Google Scholar 

  7. Wakabayashi S, Bertrand B, Shigekawa M, Fafournoux P, Pouyssegur J (1994) Growth factor activation and “H(+)-sensing” of the Na+/H+ exchanger isoform 1 (NHE1). Evidence for an additional mechanism not requiring direct phosphorylation. J Biol Chem 269:5583–5588

    PubMed  CAS  Google Scholar 

  8. Kiviluoto T, Paimela H, Mustonen H, Kivilaakso E (1990) Intracellular pH in isolated Necturus antral mucosa exposed to luminal acid. Gastroenterology 98:901–908

    PubMed  CAS  Google Scholar 

  9. Kiviluoto T, Mustonen H, Salo J, Kivilaakso E (1995) Regulation of intracellular pH in isolated Necturus gastric mucosa during short-term exposure to luminal acid. Gastroenterology 108:999–1004

    Article  PubMed  CAS  Google Scholar 

  10. Yanaka A, Suzuki H, Shibahara T, Matsui H, Nakahara A, Tanaka N (2002) EGF promotes gastric mucosal restitution by activating Na(+)/H(+) exchange of epithelial cells. Am J Physiol Gastrointest Liver Physiol 282:G866–G876

    PubMed  CAS  Google Scholar 

  11. Wakabayashi S, Ikeda T, Iwamoto T, Pouyssegur J, Shigekawa M (1997) Calmodulin-binding autoinhibitory domain controls “pH-sensing” in the Na+/H+ exchanger NHE1 through sequence-specific interaction. Biochemistry 36:12854–12861

    Article  PubMed  CAS  Google Scholar 

  12. Ranta-Knuuttila T, Kiviluoto T, Mustonen H, Puolakkainen P, Watanabe S, Sato N, Kivilaakso E (2002) Migration of primary cultured rabbit gastric epithelial cells requires intact protein kinase C and Ca2+/calmodulin activity. Dig Dis Sci 47:1008–1014

    Article  PubMed  CAS  Google Scholar 

  13. Grynkiewicz G, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    PubMed  CAS  Google Scholar 

  14. Takahashi A, Camacho P, Lechleiter JD, Herman B (1999) Measurement of intracellular calcium. Physiol Rev 79:1089– 1125

    PubMed  CAS  Google Scholar 

  15. Marlink KL, Bacon KD, Sheppard BC, Ashktorab H, Smoot DT, Cover TL, Deveney CW, Rutten MJ (2003) Effects of Helicobacter pylori on intracellular Ca2+ signaling in normal human gastric mucous epithelial cells. Am J Physiol Gastrointest Liver Physiol 285:G163–G176

    PubMed  CAS  Google Scholar 

  16. Mustonen H, Kiviluoto T, Paimela H, Puolakkainen P, Kivilaakso E (2005) Calcium signaling is involved in ethanol induced volume decrease and gap junction closure in cultured rat gastric mucosal cells. Dig Dis Sci 50:103–110

    Article  PubMed  CAS  Google Scholar 

  17. Mustonen H, Kiviluoto T, Puolakkainen P, Kivilaakso E (2004) Ethanol induces volume changes and gap junction closure via intracellular Ca2+ signalling pathway in cultured rabbit gastric epithelial cells. Scand J Gastroenterol 39:104–110

    Article  PubMed  CAS  Google Scholar 

  18. Athmann C, Zeng N, Scott DR, Sachs G (2000) Regulation of parietal cell calcium signaling in gastric glands. Am J Physiol Gastrointest Liver Physiol 279:G1048–G1058

    PubMed  CAS  Google Scholar 

  19. Thomas JA, Buchsbaum RN, Zimniak A, Racker E (1979) Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ. Biochemistry 18:2210–2218

    Article  PubMed  CAS  Google Scholar 

  20. Masereel B, Pochet L, Laeckmann D (2003) An overview of inhibitors of Na(+)/H(+) exchanger. Eur J Med Chem 38:547–554

    Article  PubMed  CAS  Google Scholar 

  21. Nylander-Koski O, Mustonen H, Kiviluoto T, Kivilaakso E (2005) Cell volume regulation during hyperosmotic shrinkage is mediated By Na+/K+-ATPase and Na+K+2Cl− cotransporter in necturus gastric surface epithelial cells. Dig Dis Sci 50(11), 2043–2049

    Google Scholar 

  22. Paris S, Pouyssegur J (1984) Growth factors activate the Na+/H+ antiporter in quiescent fibroblasts by increasing its affinity for intracellular H+. J Biol Chem 259:10989–10994

    PubMed  CAS  Google Scholar 

  23. Bianchini L, L’Allemain G, Pouyssegur J (1997) The p42/p44 mitogen-activated protein kinase cascade is determinant in mediating activation of the Na+/H+ exchanger (NHE1 isoform) in response to growth factors. J Biol Chem 272:271–279

    Article  PubMed  CAS  Google Scholar 

  24. Phan VN, Kusuhara M, Lucchesi PA, Berk BC (1997) A 90-kD Na(+)-H+ exchanger kinase has increased activity in spontaneously hypertensive rat vascular smooth muscle cells. Hypertension 29:1265–1272

    PubMed  CAS  Google Scholar 

  25. Wakabayashi S, Bertrand B, Ikeda T, Pouyssegur J, Shigekawa M (1994) Mutation of calmodulin-binding site renders the Na+/H+ exchanger (NHE1) highly H(+)-sensitive and Ca2+ regulation-defective. J Biol Chem 269:13710–13715

    PubMed  CAS  Google Scholar 

  26. Wakabayashi S, Ikeda T, Noel J, Schmitt B, Orlowski J, Pouyssegur J, Shigekawa M (1995) Cytoplasmic domain of the ubiquitous Na+/H+ exchanger NHE1 can confer Ca2+ responsiveness to the apical isoform NHE3. J Biol Chem 270:26460– 26465

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by the Research Foundation of Helsinki University Central Hospital, the Academy of Finland, and the Jenny and Antti Wihuri Foundation, Helsinki, Finland. The authors thank Mrs Paula Kokko for excellent technical assistance.

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Authors and Affiliations

  1. Department of Gastrointestinal Surgery, Helsinki University Central Hospital, Helsinki, Finland

    Outi Nylander-Koski, Harri Mustonen, Pauli Puolakkainen, Tuula Kiviluoto & Eero Kivilaakso

  2. Department of Surgery, Helsinki University Central Hospital, Biomedicum Helsinki, Haartmaninkatu 8, 00290, Helsinki, Finland

    Harri Mustonen

Authors
  1. Outi Nylander-Koski
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  2. Harri Mustonen
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  3. Pauli Puolakkainen
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  4. Tuula Kiviluoto
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  5. Eero Kivilaakso
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Correspondence to Harri Mustonen.

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Nylander-Koski, O., Mustonen, H., Puolakkainen, P. et al. Epidermal Growth Factor Enhances Intracellular pH Regulation via Calcium Signaling in Acid-Exposed Primary Cultured Rabbit Gastric Epithelial Cells. Dig Dis Sci 51, 1322–1330 (2006). https://doi.org/10.1007/s10620-006-9075-7

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  • DOI: https://doi.org/10.1007/s10620-006-9075-7

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