Abstract
The effect of ANG II on pHi, [Ca2+]i and cell volume was investigated in T84 cells, a cell line originated from colon epithelium, using the probes BCECF-AM, Fluo 4-AM and acridine orange, respectively. The recovery rate of pHi via the Na+/H+ exchanger was examined in the first 2 min following the acidification of pHi with a NH4Cl pulse. In the control situation, the pHi recovery rate was 0.118 ± 0.001 (n = 52) pH units/min and ANG II (10−12 M or 10−9 M) increased this value (by 106% or 32%, respectively) but ANG II (10−7 M) decreased it to 47%. The control [Ca2+]i was 99 ± 4 (n = 45) nM and ANG II increased this value in a dose-dependent manner. The ANG II effects on cell volume were minor and late and should not interfere in the measurements of pHi recovery and [Ca2+]i. To document the signaling pathways in the hormonal effects we used: Staurosporine (a PKC inhibitor), W13 (a calcium-dependent calmodulin antagonist), H89 (a PKA inhibitor) or Econazole (an inhibitor of cytochrome P450 epoxygenase). Our results indicate that the biphasic effect of ANG II on Na+/H+ exchanger is a cAMP-independent mechanism and is the result of: 1) stimulation of the exchanger by PKC signaling pathway activation (at 10−12 – 10−7 M ANG II) and by increases of [Ca2+]i in the lower range (at 10−12 M ANG II) and 2) inhibition of the exchanger at high [Ca2+]i levels (at 10−9 – 10−7 M ANG II) through cytochrome P450 epoxygenase-dependent metabolites of the arachidonic acid signaling pathway.
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References
Binder H.J., Sandle G.E. 1994. Electrolyte transport in the mammalian colon. In: L.R. Johnson, editor. Physiology of the Gastrointestinal Tract. Raven Press, New York pp 2133–2171
Binder H.J. 2003. Intestinal fluid and electrolyte movement. In: W.F. Boron, E.L. Boulpaep, editors. Medical Physiology. Saunders, Philadelphia pp 930–946
Boron W.F., Weer P. 1976. Intracellular pH transients in squid giant axons caused by CO2, NH3, and metabolic inhibitors. J. Gen. Physiol. 67:91–112
Cano A., Miller R.T., Alpern R.J., Preisig P.A. 1994. Angiotensin II stimulation of Na-H antiporter activity is cAMP independent in OKP cells. Am. J. Physiol. 266:C1603–C1608
Devor D.C., Frizzell R.A. 1998. Modulation of K+ channels by arachidonic acid in T84 cells. Inhibition of Ca2+-dependent K+ channel. Am. J. Physiol. 274:C138–C148
Dharmsathaphorn K., McRoberts J.A., Mandel K.G., Tisdale L.D., Masui H. 1984. A human colonic tumor cell line that maintains vectorial electrolyte transport. Am. J. Physiol. 246:G204–G208
Douglas J.G., Hopper U. 1994. Novel aspects of angiotensin receptors and signal transduction in the kidney. Annu. Rev. Physiol. 56:649–669
Du Z., Ferguson W., Wang T. 2003. Role of PKC and calcium in modulation of effects of angiotensin II on sodium transport in proximal tubule. Am. J. Physiol. 284:F688–F692
Foster E., Dudeja P., Brasitus T. 1986. Na+/H+ exchange in rat colonic brush-border membrane vesicles. Am. J. Physiol. 250:G781–G787
Grynkiewicz G., Poenie M., Tsien R.Y. 1985. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J. Biol. Chem. 260:3440–3450
Harris P.J., Young J.A. 1977. Dose-dependent stimulation and inhibition of proximal tubular sodium reabsorption by angiotensin II in the rat kidney. Pfluegers Arch. 367:295–297
Hecht G., Hodges K., Gill R.K., Kear F., Tyagi S., Malakooti J., Ramaswamy K., Dudeja P.K. 2004. Differential regulation of Na+/H+ exchange isoform activities by enteropathogenic E. coli in human intestinal epithelial cells. Am. J. Physiol. 287:G370–G378
Hirasawa K., Sato Y., Hosoda Y., Yamamoto T., Hanai H. 2002. Immunohistochemical localization of angiotensin II receptor and local renin-angiotensin system in human colonic mucosa. J. Histochem. Cytochem. 50:275–282
Houillier P., Chambrey R., Achard J.M., Froissart M., Poggioli J., Paillard M. 1996. Signaling pathways in the biphasic effect of angiotensin II on apical Na+/H+ antiport activity in proximal tubule. Kidney Int. 50:1496–1505
Liu F.Y., Cogan M.G. 1987. Angiotensin II: a potent regulator of acidification in the rat early proximal convoluted tubule. J. Clin. Invest. 80:272–275
Musa-Aziz R., Mello-Aires M. 2002. Angiotensin II modulates Na+/H+ exchanger in epithelial colon cells. J. Am. Soc. Nephrol 13:SU–P0052
Musa-Aziz, R., Mello-Aires, M. 2005. Action of ANG II and ANP on colon epithelial cells. Pfluegers Arch. in press
Navar L.G., Harrison-Bernard L.M., Wang C.T., Cervenka L., Mitchell K.D. 1999. Concentrations and actions of intraluminal angiotensin II. J. Am. Soc. Nephrol. 10:S189–S195
Oliveira-Souza M., Mello-Aires M. 2000. Interaction of angiotensin II and atrial natriuretic peptide on pHi regulation in MDCK cells. Am. J. Physiol. 279:F944–F953
Oliveira-Souza M., Musa-Aziz R., Malnic G., Mello-Aires M. 2004. Arginine vasopressin stimulates H+-ATPase in MDCK cells via VI (cell Ca2+) and V2 (cAMP) receptors. Am. J. Physiol. 286:F402–F408
Phillips M.I., Speakman E.A., Kimura B. 1993. Levels of angiotensin and molecular biology of the tissue renin angiotensin systems. Regul. Pept. 43:1–20
Poggioli J., Lazar G., Houillier P., Gardin J.P., Achard J.M., Paillard M. 1992. Effects of angiotensin II and non peptide receptor antagonists on transduction pathways in rat proximal tubule. Am. J. Physiol. 263:C750–C758
Pouysségur J. 1994. Molecular biology and hormonal regulation of vertebrate Na+/H+ exchanger isoforms. Renal Physiol. Biochem. 17:190–193
Ramirez, M.A., Beltran, A.R., Malnic, G., Rebouças, N.A. 2003. Kinetics of an apical Na+/H+ exchanger in T84 colon cells: effect of heat-stable E. coli enterotoxin (STA) (abstract). World Congress of Nephrology, M3
Ramirez M.A., Toriano R., Parisi M., Malnic G. 2000. Control of cell pH in the T84 colon cell line. J. Membrane Biol. 177:149–157
Reilly A.M., Harris P.J., Williams D.A. 1995. Biphasic effect of angiotensin II on intracellular sodium concentration in rat proximal tubules. Am. J. Physiol. 269:F374–F380
Romero M.F., Hopfer U., Madhun Z.T., Zhou W., Douglas J.G. 1991. Angiotensin II actions in the rabbit proximal tubule. Renal Physiol. Biochem. 14:199–207
Sechi L.A., Valentin J.P., Griffin C.A., Schambelan M. 1993. Autoradiographic characterization of angiotensin II receptor subtypes in rat intestine. Am. J. Physiol. 265:G21–G27
Tararthuch A.L., Fernandez R., Ramirez M.A., Malnic G. 2002. Factors affecting ammonium uptake by C11 clone of MDCK cells. Pfluegers Arch. 445:194–201
Wakabayashi S., Bertrand B., Ikeda T., Pouysségur 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
Wakabayashi S., Shigekawa M., Pouysségur J. 1997. Molecular physiology of vertebrate Na+/H+ exchangers. Physiol. Rev. 77:51–74
Weinman E.J., Hanley R., Morell G., Yuan N., Steplock D., Bui G., Shenolikar S. 1992. Regulation of the renal Na+-H+ exchanger by calcium calmodulin-dependent multifunctional protein kinase II. Miner. Electrol. Metab. 18:35–39
Weintraub W.H., Machen T.E. 1989. pH regulation in hepatoma cells: roles for Na-H exchange, Cl-HCO3 exchange, and Na-HCO3 cotransport. Am. J. Physiol. 257:G317–G327
Acknowledgments
This work was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP), Programa de Apoio à Núcleos de Excelência (PRONEX) and Conselho Nacional de Pesquisas (CNPq). The authors thank Dr. Gerhard Malnic for careful reading of the manuscript.
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Musa-Aziz, R., Oliveira-Souza, M. & Mello-Aires, M. Signaling Pathways in the Biphasic Effect of ANG II on Na+/H+ Exchanger in T84 Cells. J Membrane Biol 205, 49–60 (2005). https://doi.org/10.1007/s00232-005-0762-7
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DOI: https://doi.org/10.1007/s00232-005-0762-7