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

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.

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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 71:736–746

    PubMed  CAS  Article  Google 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–F925

    Google 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 246:330–336

    PubMed  Article  CAS  Google 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–G1093

    CAS  Google 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–F299

    CAS  Google 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 269:29704–29709

    PubMed  CAS  Google 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 72:248–254

    PubMed  Article  CAS  Google 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–F717

    PubMed  CAS  Google 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–F843

    PubMed  CAS  Google 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) 513:341–357

    Article  CAS  Google 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–G56

    PubMed  CAS  Google 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 13:9808–9816

    Article  Google Scholar 

  13. McLean IW, Nakane PF (1974) Periodate-lysine paraformaldehyde fixative: a new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083

    PubMed  CAS  Google Scholar 

  14. Orlowski J, Grinstein S (2004) Diversity of the mammalian sodium/proton exchanger SLC9 gene family. Pflügers Arch-Eur J Physiol 447:549–565

    Article  CAS  Google Scholar 

  15. Orlowski J, Kandasamy RA, Shull GE (1992) Molecular cloning and putative members of the Na+/H+ exchange gene family. J Biol Chem 267:9331–9339

    PubMed  CAS  Google 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–G478

    Google 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 276:27042–27050

    PubMed  Article  CAS  Google 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–F517

    CAS  Google Scholar 

  19. Rickmann M, Wolff JR (1995) S100 protein expression in subpopulations of neurons of rat brain. Neuroscience 67:977–991

    PubMed  Article  CAS  Google 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 272:287–294

    PubMed  Article  CAS  Google Scholar 

  21. Roussa E (2001) H+ and HCO3 transporters in human salivary glands. An immunohistochemical study. Histochem J 33:337–344

    PubMed  Article  CAS  Google Scholar 

  22. Roussa E, Thévenod F (1998) Distribution of V-ATPase in rat salivary glands. Eur J Morphol 36 (Suppl):147–152

    PubMed  Google 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 46:91–100

    PubMed  CAS  Google 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–G1296

    Google 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 49:463–474

    PubMed  CAS  Google 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 51:125–137

    PubMed  Article  CAS  Google 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–G440

    Google 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 269:24410–24417

    PubMed  Google 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–F59

    CAS  Google 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 271:32749–32752

    PubMed  Article  CAS  Google Scholar 

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Acknowledgements

The authors thank Dr. P. S. Aronson (Section of Nephrology, Department of Internal Medicine, Yale University School of Medicine, New Haven, Conn.) and Dr. S. Grinstein (Division of Cell Biology, Hospital for Sick Children, Toronto, Canada) for providing the anti-NHE4 antibodies, and Karen Fricke for excellent technical assistance.

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Correspondence to Eleni Roussa.

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This study was supported by a grant from the Deutsche Forschungsgemeinschaft (RO24495/1–1).

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Oehlke, O., Sprysch, P., Rickmann, M. et al. Na+/H+ exchanger isoforms are differentially regulated in rat submandibular gland during acid/base disturbances in vivo. Cell Tissue Res 323, 253–262 (2006). https://doi.org/10.1007/s00441-005-0055-6

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Keywords

  • Salivary glands
  • Submandibular
  • Acidosis
  • Alkalosis
  • Rat (Wistar, male)