Abstract
The uptake of adenosine in brush border vesicles of the proximal tubule of the rat kidney has been studied with a filtration technique. The initial rate of uptake was almost 6 times greater in the presence of NaCl than in the presence of KCl. The stimulatory effect of Na+ was strictly dependent on a gradient of Na+ (out>in). The time course of uptake showed an overshoot with a maximum at 20 s with a gradient of NaCl, but not with KCl. Inosine and 5′-AMP were produced from adenosine within the vesicles. In the presence of an inhibitor or adenosine deaminase adenosine was not significantly metabolized during the first 20 s of uptake. Thus, kinetic parameters of transport could be studied in the absence of interferences with metabolism. AK m of 1.1 μM and aV max of 232 pmol · min−1 · mg protein−1 were calculated for the Na+ gradient-dependent transport. The dependency on a Na+ gradient, the capacity for uphill transport and the high affinity for adenosine situate this transport system apart from the mechanisms of transport of nucleosides described so far. It may be relevant in regard to the role of adenosine in the regulation of glomerular filtration.
Similar content being viewed by others
Abbreviations
- EHNA:
-
erythro-9-(2-hydroxy-3-nonyl)adenine
- FCCP:
-
carbonylcyanide p-trifluoromethoxy-phenylhydrazone
- HEPES:
-
N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid
- Tris:
-
tris (hydroxymethyl)-aminomethane
References
Angielski S, Le Hir M, Dubach UC (1983) Transport of adenosine by renal brush border membranes. Pflügers Arch 397:75–77
Aronson PS, Saktor B (1975) The Na+ gradient-dependent transport ofD-glucose in renal brush border membranes. J Biol Chem 250:6032–6039
Barberis C, Minn A, Gayet J (1981) Adenosine transport into guinea pig synaptosomes. J Neurochem 36:341–354
Bender AS, Wu PH, Phillis JW (1980) The characterization of (3H) adenosine uptake into rat cerebral cortical synaptosomes. J Neurochem 35:629–640
Bender AS, Wu PH, Phillis JW (1981)The rapid uptake and release of (3H) adenosine by rat cerebral cortical synaptosomes. J Neurochem 36:651–660
Berlin RD (1973) Temperature dependence of nucleoside membrane transport in rabbit alveolar macrophages and polymorphonuclear leucocytes. J Biol Chem 248:4724–4730
Berlin RD, Oliver JM (1975) Membrane transport of purine and pyrimidine bases and nucleotides in animal cells. Int Rev Cytol 42:287–336
Berner W, Kinne R, Murer H (1976) Phosphate transport into brush border membrane vesicles isolated from rat small intestine. Biochem J 160:467–474
Biber J, Stieger B, Haase W, Murer H (1981) A high yield preparation of rat kidney brush border membranes. Different behaviour of lysosomal markers. Biochim Biophys Acta 647:169–176
Chello PL, Sirotnak FM, Dorick DM, Yang CH, Montgomery JA (1983) Intial rate kinetics and evidence for duality and mediated transport of adenosine, related purine nucleosides, and nucleoside analogues in L 1210 cells. Cancer Res 43:97–103
Fleit H, Conklyn M, Stebbins RD, Slber R (1975) Function of 5′-nucleotidase in the uptake of adenosine from AMP by human lymphocytes. J Biol Chem 250:8889–8892
Green RD (1980) Adenosine transport by a variant of C 1300 murine neuroblastoma cells deficient in adenosine kinase. Biochim Biophys Acta 598:366–374
Harley ER, Paterson RP, Cass CE (1982) Initial rate kinetics of the transport of adenosine and 4-amino-7(beta-D-ribofuranasyl)-pyrrolol[2,3-d] pyrimidin (tubercidin) in cultured cells. Cancer Res 42:1289–1295
Hertz L (1978) Kinetics of adenosine uptake into astrocytes. J Neurochem 31:55–62
Kinsella JL, Aronson PS (1980) Properties of the Na+−K+ exchanger in renal microvillus membrane vesicles. Am J Physiol 241:F374-F379
Klabunde RE, Althouse DG (1981) Adenosine metabolism in dog whole blood: effect of dipyridamole. Life Sci 28:2631–2641
Kuttesch JF, Nelson JA (1982) Renal handling of 2′-deoxy-adenosine and adenosine in humans and mice. Cancer Chemother Pharmacol 8:221–229
Li CC, Hochstadt J (1976) Membrane-associated enzymes involved in nucleoside processing by plasma membrane vesicles isolated from L 929 cells grown in defined medium. J Biol Chem 1181–1187
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 265–275
Lum CT, Marz R, Plagemann PGW, Wohlhueter RM (1979) Adenosine transport and metabolism in mouse leukemia cells and in canine thymocytes and peripheral blood leucocytes. J Cell Physiol 101:173–200
Manfredi JP, Sparks HV (1982) Adenosine's role in coronary vasodilation induced by atrial pacing and norepinephrine. Am J Physiol 243:H536-H545
Meunier FM, Morel N (1978) Adenosine uptake by cholinergic synaptosomes from torpedo electric organ. J. Neurochem 31:845–851
Osswald H, Hermes HH, Nabakowski G (1982) Role of adenosine in signal transmission of tubuloglomerular feedback. Kidney Int (Suppl) 12:136–143
Plagemann PGW, Wohlhueter RM (1980) Permeation of nucleosides, nucleic acid bases and nucleotides in animal cells. Curr Top Membr Transp 14:225–330
Plagemann PGW, Wohlhueter RM (1983) Adenosine and tubercidine binding and transport in Chinese Hamster ovary and Novikoff hepatoma cells. J Cell Physiol 116:247–255
Plagemann PGW, Wohlhueter RM, Erbe J (1982) Nucleoside transport in human erythrocytes. A simple carrier with directional symmetry and differential mobility of loaded and empty carrier. J Biol Chem 257:12069–12074
Rubio R, Berne RM (1980) Localization of purine and pyrimidine nucleoside phosphorylases in heart, kidney and liver. Am J Physiol 239:4721–4730
Sato T, Kuninaka A, Yoshino H, Ui M (1982) A sensitive radioimmune assay for adenosine in biological samples. Anal Biochem 121:409–420
Slowiaczeck P, Tattersall MHN (1982) The determination of purine levels in human and mouse plasma. Anal Biochem 125:6–12
Spielman WS, Thompson CI (1982) A proposed role for adenosine in the regulation of renal hemodynamics and renine release. Am J Physiol 242:F423-F435
Strauss PR, Sheehan J, Kashket E (1976) Membrane transport by murine lymophocytes. I. A rapid sampling technique as applied to the adenosine and thymidine systems. J Exp Med 144:1009–1021
Taube RA, Berlin RD (1972) Membrane transport of nucleosides in rabbit polymorphonuclear leucocytes. Biochim Biophys Acta 255:6–18
Thampy G, Barnes EM (1983a) Adenosine transport by primary cultures of neurons from chick embryo brain. J Neurochem 40:874–879
Thampy G, Barnes EM (1983b) Adenosine transport by cultured glyal cells from chick embryo brain. Arch Biochem Biophys 220:340–346
Wohlhueter RM, Plagemann PGW (1982) On the functional symmetry of nucleoside transport in mammalian cells. Biochim Biophys Acta 689:249–260
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Le Hir, M., Dubach, U.C. Sodium gradient-energized concentrative transport of adenosine in renal brush border vesicles. Pflugers Arch. 401, 58–63 (1984). https://doi.org/10.1007/BF00581533
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00581533