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. 1987 Oct;80(4):970–978. doi: 10.1172/JCI113190

Role of the Na+/H+ antiporter in rat proximal tubule bicarbonate absorption.

P A Preisig 1, H E Ives 1, E J Cragoe Jr 1, R J Alpern 1, F C Rector Jr 1
PMCID: PMC442334  PMID: 2888788

Abstract

Amiloride and the more potent amiloride analog, 5-(N-t-butyl) amiloride (t-butylamiloride), were used to examine the role of the Na+/H+ antiporter in bicarbonate absorption in the in vivo microperfused rat proximal convoluted tubule. Bicarbonate absorption was inhibited 29, 46, and 47% by 0.9 mM or 4.3 mM amiloride, or 1 mM t-butylamiloride, respectively. Sensitivity of the Na+/H+ antiporter to these compounds in vivo was examined using fluorescent measurements of intracellular pH with (2', 7')-bis(carboxyethyl)-(5,6)-carboxyfluorescein (BCECF). Amiloride and t-butylamiloride were shown to be as potent against the antiporter in vivo as in brush border membrane vesicles. A model of proximal tubule bicarbonate absorption was used to correct for changes in the luminal profiles for pH and inhibitor concentration, and for changes in luminal flow rate in the various series. We conclude that the majority of apical membrane proton secretion involved in transepithelial bicarbonate absorption is mediated by the Na+-dependent, amiloride-sensitive Na+H+ antiporter. However, a second mechanism of proton secretion contributes significantly to bicarbonate absorption. This mechanism is Na+-independent and amiloride-insensitive.

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Selected References

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  1. Akiba T., Alpern R. J., Eveloff J., Calamina J., Warnock D. G. Electrogenic sodium/bicarbonate cotransport in rabbit renal cortical basolateral membrane vesicles. J Clin Invest. 1986 Dec;78(6):1472–1478. doi: 10.1172/JCI112738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alpern R. J., Chambers M. Cell pH in the rat proximal convoluted tubule. Regulation by luminal and peritubular pH and sodium concentration. J Clin Invest. 1986 Aug;78(2):502–510. doi: 10.1172/JCI112602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alpern R. J., Cogan M. G., Rector F. C., Jr Effect of luminal bicarbonate concentration on proximal acidification in the rat. Am J Physiol. 1982 Jul;243(1):F53–F59. doi: 10.1152/ajprenal.1982.243.1.F53. [DOI] [PubMed] [Google Scholar]
  4. Alpern R. J. Mechanism of basolateral membrane H+/OH-/HCO-3 transport in the rat proximal convoluted tubule. A sodium-coupled electrogenic process. J Gen Physiol. 1985 Nov;86(5):613–636. doi: 10.1085/jgp.86.5.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Alpern R. J., Rector F. C., Jr A model of proximal tubular bicarbonate absorption. Am J Physiol. 1985 Feb;248(2 Pt 2):F272–F281. doi: 10.1152/ajprenal.1985.248.2.F272. [DOI] [PubMed] [Google Scholar]
  6. Aronson P. S., Nee J., Suhm M. A. Modifier role of internal H+ in activating the Na+-H+ exchanger in renal microvillus membrane vesicles. Nature. 1982 Sep 9;299(5879):161–163. doi: 10.1038/299161a0. [DOI] [PubMed] [Google Scholar]
  7. Aronson P. S., Suhm M. A., Nee J. Interaction of external H+ with the Na+-H+ exchanger in renal microvillus membrane vesicles. J Biol Chem. 1983 Jun 10;258(11):6767–6771. [PubMed] [Google Scholar]
  8. Berry C. A. Electrical effects of acidification in the rabbit proximal convoluted tubule. Am J Physiol. 1981 May;240(5):F459–F470. doi: 10.1152/ajprenal.1981.240.5.F459. [DOI] [PubMed] [Google Scholar]
  9. Biagi B. A. Effects of the anion transport inhibitor, SITS, on the proximal straight tubule of the rabbit perfused in vitro. J Membr Biol. 1985;88(1):25–31. doi: 10.1007/BF01871210. [DOI] [PubMed] [Google Scholar]
  10. Boron W. F., Boulpaep E. L. Intracellular pH regulation in the renal proximal tubule of the salamander. Basolateral HCO3- transport. J Gen Physiol. 1983 Jan;81(1):53–94. doi: 10.1085/jgp.81.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Burg M., Green N. Bicarbonate transport by isolated perfused rabbit proximal convoluted tubules. Am J Physiol. 1977 Oct;233(4):F307–F314. doi: 10.1152/ajprenal.1977.233.4.F307. [DOI] [PubMed] [Google Scholar]
  12. Burg M., Patlak C., Green N., Villey D. Organic solutes in fluid absorption by renal proximal convoluted tubules. Am J Physiol. 1976 Aug;231(2):627–637. doi: 10.1152/ajplegacy.1976.231.2.627. [DOI] [PubMed] [Google Scholar]
  13. Burnham C., Munzesheimer C., Rabon E., Sachs G. Ion pathways in renal brush border membranes. Biochim Biophys Acta. 1982 Mar 8;685(3):260–272. doi: 10.1016/0005-2736(82)90066-9. [DOI] [PubMed] [Google Scholar]
  14. Cassano G., Stieger B., Murer H. Na/H- and Cl/OH-exchange in rat jejunal and rat proximal tubular brush border membrane vesicles. Studies with acridine orange. Pflugers Arch. 1984 Mar;400(3):309–317. doi: 10.1007/BF00581565. [DOI] [PubMed] [Google Scholar]
  15. Chan Y. L., Giebisch G. Relationship between sodium and bicarbonate transport in the rat proximal convoluted tubule. Am J Physiol. 1981 Mar;240(3):F222–F230. doi: 10.1152/ajprenal.1981.240.3.F222. [DOI] [PubMed] [Google Scholar]
  16. Chantrelle B., Cogan M. G., Rector F. C., Jr Evidence for coupled sodium/hydrogen exchange in the rat superficial proximal convoluted tubule. Pflugers Arch. 1982 Nov 11;395(3):186–189. doi: 10.1007/BF00584807. [DOI] [PubMed] [Google Scholar]
  17. Cragoe E. J., Jr, Woltersdorf O. W., Jr, Bicking J. B., Kwong S. F., Jones J. H. Pyrazine diuretics. II. N-amidino-3-amino-5-substituted 6-halopyrazinecarboxamides. J Med Chem. 1967 Jan;10(1):66–75. doi: 10.1021/jm00313a014. [DOI] [PubMed] [Google Scholar]
  18. Frömter E., Gessner K. Active transport potentials, membrane diffusion potentials and streaming potentials across rat kidney proximal tubule. Pflugers Arch. 1974;351(1):85–98. doi: 10.1007/BF00603513. [DOI] [PubMed] [Google Scholar]
  19. Gurich R. W., Warnock D. G. Electrically neutral Na+-H+ exchange in endosomes obtained from rabbit renal cortex. Am J Physiol. 1986 Oct;251(4 Pt 2):F702–F709. doi: 10.1152/ajprenal.1986.251.4.F702. [DOI] [PubMed] [Google Scholar]
  20. Howlin K. J., Alpern R. J., Rector F. C., Jr Amiloride inhibition of proximal tubular acidification. Am J Physiol. 1985 Jun;248(6 Pt 2):F773–F778. doi: 10.1152/ajprenal.1985.248.6.F773. [DOI] [PubMed] [Google Scholar]
  21. Ives H. E., Yee V. J., Warnock D. G. Asymmetric distribution of the Na+/H+ antiporter in the renal proximal tubule epithelial cell. J Biol Chem. 1983 Nov 25;258(22):13513–13516. [PubMed] [Google Scholar]
  22. Ives H. E., Yee V. J., Warnock D. G. Mixed type inhibition of the renal Na+/H+ antiporter by Li+ and amiloride. Evidence for a modifier site. J Biol Chem. 1983 Aug 25;258(16):9710–9716. [PubMed] [Google Scholar]
  23. Jacobson H. R., Kokko J. P. Intrinsic differences in various segments of the proximal convoluted tubule. J Clin Invest. 1976 Apr;57(4):818–825. doi: 10.1172/JCI108357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kinne-Saffran E., Beauwens R., Kinne R. An ATP-driven proton pump in brush-border membranes from rat renal cortex. J Membr Biol. 1982;64(1-2):67–76. doi: 10.1007/BF01870769. [DOI] [PubMed] [Google Scholar]
  25. Kinne-Saffran E., Kinne R. Further evidence for the existence of an intrinsic bicarbonate-stimulated Mg2+-ATPase in brush border membranes isolated from rat kidney cortex. J Membr Biol. 1979 Sep;49(3):235–251. doi: 10.1007/BF01871120. [DOI] [PubMed] [Google Scholar]
  26. Kinne-Saffran E., Kinne R. Presence of bicarbonate stimulated ATPase in the brush border microvillus membranes of the proximal tubule. Proc Soc Exp Biol Med. 1974 Jul;146(3):751–753. doi: 10.3181/00379727-146-38186. [DOI] [PubMed] [Google Scholar]
  27. Kinsella J. L., Aronson P. S. Amiloride inhibition of the Na+-H+ exchanger in renal microvillus membrane vesicles. Am J Physiol. 1981 Oct;241(4):F374–F379. doi: 10.1152/ajprenal.1981.241.4.F374. [DOI] [PubMed] [Google Scholar]
  28. Kinsella J. L., Freiberg J. M., Sacktor B. Glucocorticoid activation of Na+/H+ exchange in renal brush border vesicles: kinetic effects. Am J Physiol. 1985 Feb;248(2 Pt 2):F233–F239. doi: 10.1152/ajprenal.1985.248.2.F233. [DOI] [PubMed] [Google Scholar]
  29. Kinsella J., Sacktor B. Thyroid hormones increase Na+-H+ exchange activity in renal brush border membranes. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3606–3610. doi: 10.1073/pnas.82.11.3606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Liang C. T., Sacktor B. Bicarbonate-stimulated ATPase in the renal proximal tubule luminal (brush border) memebrane. Arch Biochem Biophys. 1976 Sep;176(1):285–297. doi: 10.1016/0003-9861(76)90167-3. [DOI] [PubMed] [Google Scholar]
  31. McKinney T. D., Burg M. B. Biocarbonate and fluid absorption by renal proximal straight tubules. Kidney Int. 1977 Jul;12(1):1–8. doi: 10.1038/ki.1977.72. [DOI] [PubMed] [Google Scholar]
  32. Murer H., Hopfer U., Kinne R. Sodium/proton antiport in brush-border-membrane vesicles isolated from rat small intestine and kidney. Biochem J. 1976 Mar 15;154(3):597–604. [PMC free article] [PubMed] [Google Scholar]
  33. Preisig P. A., Berry C. A. Evidence for transcellular osmotic water flow in rat proximal tubules. Am J Physiol. 1985 Jul;249(1 Pt 2):F124–F131. doi: 10.1152/ajprenal.1985.249.1.F124. [DOI] [PubMed] [Google Scholar]
  34. Sabolić I., Burckhardt G. Characteristics of the proton pump in rat renal cortical endocytotic vesicles. Am J Physiol. 1986 May;250(5 Pt 2):F817–F826. doi: 10.1152/ajprenal.1986.250.5.F817. [DOI] [PubMed] [Google Scholar]
  35. Sabolić I., Haase W., Burckhardt G. ATP-dependent H+ pump in membrane vesicles from rat kidney cortex. Am J Physiol. 1985 Jun;248(6 Pt 2):F835–F844. doi: 10.1152/ajprenal.1985.248.6.F835. [DOI] [PubMed] [Google Scholar]
  36. Sasaki S., Berry C. A., Rector F. C., Jr Effect of potassium concentration on bicarbonate reabsorption in the rabbit proximal convoluted tubule. Am J Physiol. 1983 Feb;244(2):F122–F128. doi: 10.1152/ajprenal.1983.244.2.F122. [DOI] [PubMed] [Google Scholar]
  37. Schwartz G. J., Al-Awqati Q. Carbon dioxide causes exocytosis of vesicles containing H+ pumps in isolated perfused proximal and collecting tubules. J Clin Invest. 1985 May;75(5):1638–1644. doi: 10.1172/JCI111871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ullrich K. J., Capasso G., Rumrich G., Papavassiliou F., Klöss S. Coupling between proximal tubular transport processes. Studies with ouabain, SITS and HCO3-free solutions. Pflugers Arch. 1977 Apr 25;368(3):245–252. doi: 10.1007/BF00585203. [DOI] [PubMed] [Google Scholar]
  39. Ullrich K. J., Rumrich G., Baumann K. Renal proximal tubular buffer-(glycodiazine) transport. Inhomogeneity of local transport rate, dependence on sodium, effect of inhibitors and chronic adaptation. Pflugers Arch. 1975 Jun 26;357(3-4):149–163. doi: 10.1007/BF00585971. [DOI] [PubMed] [Google Scholar]
  40. Vurek G. G., Warnock D. G., Corsey R. Measurement of picomole amounts of carbon dioxide by calorimetry. Anal Chem. 1975 Apr;47(4):765–767. doi: 10.1021/ac60354a024. [DOI] [PubMed] [Google Scholar]
  41. Wang W., Messner G., Oberleithner H., Lang F., Deetjen P. The effect of ouabain on intracellular activities of K+, Na+, Cl-, H+ and Ca2+ in proximal tubules of frog kidneys. Pflugers Arch. 1984 May;401(1):6–13. doi: 10.1007/BF00581526. [DOI] [PubMed] [Google Scholar]
  42. Warnock D. G., Reenstra W. W., Yee V. J. Na+/H+ antiporter of brush border vesicles: studies with acridine orange uptake. Am J Physiol. 1982 Jun;242(6):F733–F739. doi: 10.1152/ajprenal.1982.242.6.F733. [DOI] [PubMed] [Google Scholar]
  43. Yoshitomi K., Burckhardt B. C., Frömter E. Rheogenic sodium-bicarbonate cotransport in the peritubular cell membrane of rat renal proximal tubule. Pflugers Arch. 1985 Dec;405(4):360–366. doi: 10.1007/BF00595689. [DOI] [PubMed] [Google Scholar]

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