Abstract
The quenching of the acridine orange fluorescence was used to monitor the formation and/or dissipation of a Δ pH in brush border vesicles isolated from rat kidney cortex or rat jejunum. Similar findings were obtained with both brush border membrane vesicle preparations.
Acridine orange fluorescence was quenched by a preset Δ pH (intravesicular acid) or by the ionophore (valinomycin/CCCP) dependent development of a Δ pH (intravesicular acid) under conditions of potassium efflux.
Under sodium efflux conditions, an acidification of the intravesicular space occurred: a) due to indirect (electrical) coupling of sodium and proton fluxes; b) due to directly coupled sodium/proton exchange.
The initial rate of the dissipation of a preset Δ pH was accelerated by pulse injections of sodium in a saturable manner; lithium partially replaced sodium. The sodium dependent acceleration in the rate of dissipation of a preset Δ pH was not altered by replacing gluconate with chloride. Amiloride was an inhibitor of directly coupled sodium/proton exchange.
An inwardly directed chloride gradient did not induce intravesicular acidification. The initial rate of the dissipative proton fluxes (preset Δ pH) was slightly accelerated by an outwardly directed chloride gradient. Sodium/proton exchange dependent acidification of the intravesicular space was not altered by replacing gluconate with chloride.
These results clearly document the existence of sodium/proton exchange in both renal and intestinal brush border membrane vesicles. In contrast, Cl/OH exchange — under our experimental conditions — must have a much smaller rate than Na/H exchange.
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Abbreviations
- HEPES:
-
N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid
- MES:
-
2-[N-morpholino]ethanesulfonic acid
- Tris:
-
tris-(hydroxymethyl)aminomethane
- TMA:
-
tetramethylammonium
- CCCP:
-
carbonyl cyanide-m-chlorophenylhydrazone
References
Aronson PS, Nee J, Suhm MA (1982) Modifier role of internal, H+ in activating Na+-H+ exchanger in renal microvillus membrane vesicles. Nature 299:161–163
Aronson PS, Suhm MA, Nee J (1983) Interaction of external H+ with the Na+-H+ exchanger in renal microvillus membrane vesicles. J Biol Chem 258:6767–6771
Benos DJ (1982) Amiloride: A molecular probe of sodium transport in tissues and cells. Am J Physiol 242:C131-C145
Biber J, Stieger B, Haase W, Murer H (1981) A high yield preparation for rat kidney brush border membranes. Different behaviour of lysosomal markers. Biochim Biophys Acta 647:169–176
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 72:248–252
Burnham C, Munzesheimer C Rabou E, Sachs G (1982) Ion pathways in renal brush border vesicles. Biochim Biophys Acta 685:260–272
Dubinsky W, Frizzell RA (1982) A novel effect of amiloride on H+-dependent Na+-transport. Am J Physiol 245:C157-C159
Freiberg MJ, Kinsella J, Sacktor B (1982) Glucocorticoids increase the Na+-H+ exchange and decrease the Na+-gradient dependent phosphate uptake systems in renal brush border membrane vesicles. Proc Natl Acad Sci USA 79:4932–4936
Hauser H, Howell K, Dawson RML, Bowyer DE (1980) Rabbit small intestinal brush border membrane preparation and lipid composition. Biochim Biophys Acta 602:567–577
Kinsella JL, Aronson PS (1980) Properties of the Na+-H+ exchanger in renal microvillus membrane vesicles. Am J Physiol 238:F451-F469
Kinsella JL, Aronson PS (1981a) Amiloride inhibition of the Na+-H+ exchanger in renal microvillus membrane vesicles. Am J Physiol 241:F374-F379
Kinsella JL, Aronson PS (1981b) Interaction of Na+ and Li+ with the renal microvillus membrane Na+−H+ exchanger. Am J Physiol 241:C220-C226
Knickelbein RG, Aronson PS, Dobbins JW (1982) Na/H and Cl/HCO3 exchange in rabbit ileal brush border membrane vesicles. Fed Proc 41:1494
Knickelbein, RG, Aronson PS, Aherton W, Dobbins JW (1983) Sodium and chloride transport across rabbit ileal brush border. I. Evidence for Na−H exchange. Am J Physiol 245:G504-G510
Liedtke CM, Hopfer U (1982) Mechanism of Cl−-translocation across small intestinal brush border membranes. II. Demonstration of Cl−−OH−-exchange and Cl− conductance. Am J Physiol 242:G272-G280
Murer H, Burckhardt G (1983) Membrane transport of anions across epithelia of mammalian small intestine and kidney proximal tubule. Rev Physiol Biochem Pharmacol 96:1–51
Murer H, Hopfer U, Kinne R (1976) Sodium/proton antiport in brush border membrane vesicles isolated from rat small intestine and kidney. Biochem J 154:597–604
Nord EP, Wright E (1983) The quantitative contribution of the Na+/H+ antiporter and Na+-diffusive pathway to total Na+-flux across the luminal membrane of the proximal tubule in the absence of organic solutes. Kidney Int 23:264
Reenstra WW, Warnock DG, Yee VJ, Forte JG (1981) Proton gradients in renal cortex brush border membrane vesicles. J Biol Chem 256:11663–11666
Sabolic I, Burckhardt G (1983a) Apparent inhibition of Na+/H+ exchange by amiloride and harmaline in acridine orange studies. Biochim Biophys Acta 731:354–360
Sabolic I, Burckhardt G (1983b) Proton pathways in rat renal brush border and basolateral membranes. Biochim Biophys Acta 734:210–220
Seifter J, Knickelbein R, Aronson PS (1983) Cl-transport in rabbit renal and microvillus membrane vesicles: evidence against Cl-OH exchange. Kidney Int 23:266
Stieger B, Murer H (1983) Heterogeneity of brush-border-membrane vesicles from rat small intestine prepared by a precipitation method using Mg/EGTA. Eur J Biochem:95–101
Turner JR, Moran, A (1982) Further studies of proximal tubular brush border membrane. D-glucose transport heterogeneity. J Membr Biol 70:37–45
Warnock DG, Yee VJ (1981) Chloride uptake by brush border membrane vesicles isolated from rabbit renal cortex. Coupling to proton gradient and K diffusion potentials. J Clin Invest 67:103–115
Warnock DG, Eveloff J (1982) NaCl entry mechanisms in the luminal membrane of the renal tubule. Am J Physiol 242:F561-F574
Warnock DG, Reenstra WW, Yee VJ (1982) Na+/H+ antiporter of brush border vesicles: studies with acridine orange uptake. Am J Physiol 242:F733-F739
Wright EM, Schell RE, Krasne S (1983) Proton permeability of renal brush border membranes. Kidney Int 23:241
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Cassano, G., Stieger, B. & Murer, H. Na/H- and Cl/OH-exchange in rat jejunal and rat proximal tubular brush border membrane vesicles. Pflugers Arch. 400, 309–317 (1984). https://doi.org/10.1007/BF00581565
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DOI: https://doi.org/10.1007/BF00581565