Cell and Tissue Research

, Volume 323, Issue 2, pp 253–262

Na+/H+ exchanger isoforms are differentially regulated in rat submandibular gland during acid/base disturbances in vivo

  • Oliver Oehlke
  • Patricia Sprysch
  • Michael Rickmann
  • Eleni Roussa
Regular Article

Abstract

Acute metabolic acidosis and alkalosis cause a series of homeostatic adaptive responses in the kidney and other epithelia. We hypothesized that acid/base disturbances might affect the expression of Na+/H+ exchanger (NHE) isoforms in salivary glands and determined the expression and cellular distribution of NHE3 and NHE4 in rat submandibular glands of controls and after imposed acute or chronic metabolic acidosis or alkalosis in vivo. Reverse transcription/polymerase chain reaction, in situ hybridization, and immunohistochemistry were applied by using specific primers, antisense probes, and antibodies, respectively. The results showed NHE3 and NHE4 transcript expression and protein abundance in rat submandibular gland. NHE3 was apically localized in duct cells, whereas NHE4 was found basolaterally distributed in acinar and duct cells. Acute acidosis and alkalosis and chronic acidosis had no effect on NHE3 and NHE4 expression and localization. In contrast, chronic metabolic alkalosis significantly decreased the number of apically stained NHE3 duct cells but had no effect on NHE3 mRNA expression. The results demonstrate, for the first time, the presence of NHE4 protein in salivary glands. The data also indicate the distinct regulation and adaptive changes of different isoforms of the same transporter in rat submandibular gland as a response to acid/base disturbances.

Keywords

Salivary glands Submandibular Acidosis Alkalosis Rat (Wistar, male) 

References

  1. Alpern RJ, Cogan MG, Rector FCJ (1983) Effects of extracellular fluid volume and plasma bicarbonate concentration on proximal acidification in the rat. J Clin Invest 71:736–746PubMedCrossRefGoogle Scholar
  2. Ambühl PM, Amemiya M, Danczkay M, Lötscher M, Kaissling B, Moe OW, Preisig PA, Alpern RJ (1996) Chronic metabolic acidosis increases NHE3 protein abundance in rat kidney. Am J Physiol Renal Physiol 271:F917–F925Google Scholar
  3. Anderie I, Blum R, Haase W, Grinstein S, Thévenod F (1998) Expression of NHE1 and NHE4 in rat pancreatic zymogen granule membranes. Biochem Biophys Res Commun 246:330–336PubMedCrossRefGoogle Scholar
  4. Bachmann O, Sonnentag T, Siegel W-K, Lambrecht G, Weichert A, Gregor M, Seidler U (1998) Different acid secretagogues activate different Na+/H+ exchanger isoforms in rabbit parietal cells. Am J Physiol Gastrointest Liver Physiol 275:G1085–G1093Google Scholar
  5. Biemesderfer D, Rutherford PA, Nagy T, Pizzonia JH, Abu-Alfa AK, Aronson PS (1997) Monoclonal antibodies for high-resolution localization of NHE3 in adult and neonatal rat kidney. Am J Physiol Renal Physiol 273:F289–F299Google Scholar
  6. Bookstein C, Musch MW, DePaoli A, Xie Z, Villeral M, Rao MC, Chang EB (1994) A unique sodium/hydrogen exchange isoform (NHE-4) of the inner medulla of the rat kidney is induced by hyperosmolarity. J Biol Chem 269:29704–29709PubMedGoogle Scholar
  7. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  8. Chambrey R, St John PL, Eladari D, Quentin F, Warnock DG, Abrahamson DR, Podevin RA, Paillard M (2001) Localization and functional characterization of Na+/H+ exchanger isoform NHE4 in rat thick ascending limbs. Am J Physiol Renal Physiol 281:F707–F717PubMedGoogle Scholar
  9. Eladari D, Leviel F, Pezy F, Paillard M, Chambrey R (2002) Rat proximal NHE3 adapts to chronic acid-base disorders but not to chronic changes in dietary NaCl intake. Am J Physiol Renal Physiol 282:F835–F843PubMedGoogle Scholar
  10. Lee MG, Schultheis PJ, Yan Ming, Shull GE, Bookstein C, Chang E, Tse M, Donowitz M, Park K, Muallem S (1998) Membrane-limited expression of Na+-H+ exchanger isoforms by P2 receptors in the rat submandibular gland duct. J Physiol (Lond) 513:341–357CrossRefGoogle Scholar
  11. Lucioni A, Womack C, Musch MW, Rocha FL, Bookstein C, Chang EB (2002) Metabolic acidosis in rats increases intestinal NHE2 and NHE3 expression and function. Am J Physiol Gastrointest Liver Physiol 283:G51–G56PubMedGoogle Scholar
  12. Luo X, Choi JY, Ko SBH, Pushkin A, Kurtz I, Ahn W, Lee MG, Muallem S (2001) HCO3 salvage mechanisms in the submandibular gland acinar and duct cells. J Biol Chem 13:9808–9816CrossRefGoogle Scholar
  13. McLean IW, Nakane PF (1974) Periodate-lysine paraformaldehyde fixative: a new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083PubMedGoogle Scholar
  14. Orlowski J, Grinstein S (2004) Diversity of the mammalian sodium/proton exchanger SLC9 gene family. Pflügers Arch-Eur J Physiol 447:549–565CrossRefGoogle Scholar
  15. Orlowski J, Kandasamy RA, Shull GE (1992) Molecular cloning and putative members of the Na+/H+ exchange gene family. J Biol Chem 267:9331–9339PubMedGoogle Scholar
  16. Park K, Olschowska JA, Richardson LA, Bookstein C, Chang EB, Melvin JE (1999) Expression of multiple Na+/H+ exchanger isoforms in rat parotid acinar and ductal cells. Am J Physiol Gastrointest Liver Physiol 39:G470–G478Google Scholar
  17. Park K, Evans RL, Watson GE, Nehrke K, Richardson L, Bell SM, Schultheis PJ, Hand A, Shull GE, Melvin JE (2001) Defective fluid secretion and NaCl absorption in the parotid glands of Na+/H+ exchanger-deficient mice. J Biol Chem 276:27042–27050PubMedCrossRefGoogle Scholar
  18. Pizzonia JH, Biemesderfer D, Abu-Alfa AK, Wu M-S, Exner M, Isenring P, Igarashi P, Aronson PS (1998) Immunochemical characterization of Na+/H+ exchanger isoform NHE4. Am J Physiol Renal Physiol 275:F510–F517Google Scholar
  19. Rickmann M, Wolff JR (1995) S100 protein expression in subpopulations of neurons of rat brain. Neuroscience 67:977–991PubMedCrossRefGoogle Scholar
  20. Robertson MA, Woodside M, Foskett JK, Orlowski J, Grinstein S (1997) Muscarinic agonists induce phosphorylation-independent activation of the NHE1-isoform of the Na+/H+ antiporter in salivary acinar cells. J Biol Chem 272:287–294PubMedCrossRefGoogle Scholar
  21. Roussa E (2001) H+ and HCO3 transporters in human salivary glands. An immunohistochemical study. Histochem J 33:337–344PubMedCrossRefGoogle Scholar
  22. Roussa E, Thévenod F (1998) Distribution of V-ATPase in rat salivary glands. Eur J Morphol 36 (Suppl):147–152PubMedGoogle Scholar
  23. Roussa E, Thévenod F, Sabolic I, Herak-Kramberger CM, Nastainczyk W, Bock R, Schulz I (1998) Immunolocalization of vacuolar type H+-ATPase in rat submandibular gland and adaptive changes induced by acid-base disturbances. J Histochem Cytochem 46:91–100PubMedGoogle Scholar
  24. Roussa E, Romero MF, Schmitt BM, Boron WF, Alper SL, Thévenod F (1999) Immunolocalization of anion exchanger AE2 and Na+-HCO3 cotransporter in rat parotid and submandibular glands. Am J Physiol Gastrointest Liver Physiol 40:G1288–G1296Google Scholar
  25. Roussa E, Alper SL, Thévenod F (2001) Immunolocalization of anion exchanger AE2, Na+/H+ exchangers NHE1 and NHE4, and vacuolar type H+-ATPase in rat pancreas. J Histochem Cytochem 49:463–474PubMedGoogle Scholar
  26. Sabolic I, Brown D, Gluck SL, Alper SL (1997) Regulation of AE1 anion exchanger and H+-ATPase in rat cortex by acute metabolic acidosis and alkalosis. Kidney Int 51:125–137PubMedCrossRefGoogle Scholar
  27. Thévenod F, Schulz I (1988) H+-dependent calcium uptake into an IP3-sensitive calcium pool from rat parotid gland. Am J Physiol Gastrointest Liver Physiol 255:G429–G440Google Scholar
  28. Thévenod F, Anderie I, and Schulz I (1994) Monoclonal antibodies against MDR1 P-glycoprotein inhibit chloride conductance and label a 65-kDa protein in pancreatic zymogen granule membranes. J Biol Chem 269:24410–24417PubMedGoogle Scholar
  29. Wright PA, Knepper MA (1990) Glutamate dehydrogenase activities in microdissected rat nephron segments: effects of acid-base loading. Am J Physiol Renal Physiol 259:F53–F59Google Scholar
  30. Wu M-S, Biemesderfer D, Giebisch G, Aronson PS (1996) Role of NHE3 in mediating renal brush border Na+/H+ exchange. Adaptation to metabolic acidosis. J Biol Chem 271:32749–32752PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Oliver Oehlke
    • 1
  • Patricia Sprysch
    • 1
  • Michael Rickmann
    • 1
  • Eleni Roussa
    • 1
  1. 1.Center for Anatomy, Department of NeuroanatomyGeorg August UniversityGoettingenGermany

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