Summary
Experiments were performed in the distal tubule of the doubly-perfused kidney of Amphiuma to determine active and passive forces, involved in the transport processes of potassium, sodium and chloride. Ion-sensitive microelectrodes and conventional microelectrodes were applied to estimate intracellular ion activities, cell membrane potentials and net flux of potassium and chloride under control conditions and during inhibition of active transport. Sodium chloride cotransport, located in the luminal cell membrane is postulated, based on the following observations:
Total omission of sodium from the tubular lumen inhibits furosemide sensitive chloride reabsorption, decreases the lumen positive transepithelial potential difference and leads to a dramatic decrease of intracellular chloride. The experiments further suggest that potassium ions are involved in the sodium chloride transport system because potassium reabsorption is inhibited by furosemide and because intracellular sodium falls significantly when potassium ions are removed from the tubular fluid. Furthermore, there is experimental evidence that the luminal potassium uptake mechanism is suppressed after potassium adaptation. Under these conditions potassium transport is found to be insensitive to furosemide.
The data suggest a furosemide sensitive contransport system for sodium, chloride and potassium, operative in the luminal cell membrane. The energy for this carrier-mediated transport process is provided by the large “downhill” gradient of sodium across the luminal cell membrane which is maintained by the sodium pump located in the peritubular cell membrane.
Zusammenfassung
Experimente am distalen Tubulus der doppelt perfundierten Niere des Amphiuma wurden ausgeführt, um die aktiven und passiven Kräfte zu bestimmen, die in die Transportprozesse von Kalium, Natrium und Chlorid involviert sind. Ionen-sensitive und konventionelle Mikroelektroden wurden verwendet, um intrazelluläre Ionenaktivitäten, Zellmembranpotentiale und Kalium- und Chlorid Nettoflüsse unter Kontrollbedingungen und während Hemmung des aktiven Transports abzuschätzen.
Auf der Basis folgender Beobachtungen wird ein Natrium-Chlorid Kotransport postuliert, der in der luminalen Zellmembran lokalisiert ist: Entfernung von Natrium aus dem Tubuluslumen hemmt die Furosemid empfindliche Chloridresorption, verringert die luminal positive transepitheliale Potentialdifferenz und führt zu dramatischem Abfall des intrazellulären Chlorids. Die Experimente schlagen ferner vor, daß Kaliumionen im Natrium-Chlorid Transportsystem involviert sind, weil die Kaliumresorption durch Furosemid gehemmt wird, und weil intrazelluläres Natrium signifikant abfällt, wenn die Kaliumionen aus der Tubulusflüssigkeit entfernt werden. Weiters gibt es experimentelle Hinweise, daß nach der Kalium Adaptation der luminale Kalium-Aufnahmemechanismus unterdrückt ist. Unter diesen Bedingungen ist der Kaliumtransport unempfindiich auf Furosemid.
Die Daten schlagen ein Furosemid empfindliches Kotransport-System für Natrium, Chlorid und Kalium in der luminalen Zellmembran vor. Die Energie für diesen Carriervermittelten Transportprozeß wird von einem großen „Bergab“-Gradienten von Natrium über die luminale Zellmembran bereitgestellt, der seinerseits durch die in der peritubulären Zellmembran lokalisierte Natriumpumpe aufrechterhalten wird.
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References
Berliner RW, Kennedy TJ Jr, Hilton JG (1950) Renal mechanisms for excretion of potassium. Amer J Physiol 162:348–367
Boulpaep EL, Giebisch G (1978) Electrophysiological measurements on the renal tubule. In: Manuel Martinez-Maldonado (Hrsg) Methods in pharmac. Plen Publ Corp Vol 4B, pp 165–193
Burg MD, Green N (1973) Function of the thick ascending limb of Henle's loop. Amer J Physiol 224:659–668
Burg MD, Stoner L, Cardinal J, Green N (1973) Furosemide effect on isolated perfused tubules. Amer J Physiol 225:119–124
Burg MD, Bourdeau JE (1978) Function of the thick ascending limb of Henle's loop. In: Vogel HG and Ullrich KJ (Hrsg) New aspects of renal function. Excerpta Medica, Amsterdam, Oxford IV, p 91
Eveloff J, Kinne R, Kinne-Saffran E, Murer H, Silva P, Epstein FH, Stoff J, Kinter WB (1978) Coupled sodium and chloride transport into plasma membrane vesicles prepared from dogfish rectal gland. Pflügers Arch 378:87–92
Finkelstein FO, Hayslett JP (1975) Role of medullary Na-K-ATPase in renal potassium adaptation. Amer J Physiol 229:524–528
Fossat B, and Lahlou B (1979) The mechanism of coupled transport of sodium and chloride in isolated urinary bladder of the trout. J Physiol 294:211–222
Frizzell RA, Dugas MC, Schultz SG (1975) Sodium chloride transport by rabbit gallbladder. J Gen Physiol 65:769–795
Frizzell RA, Smith PL, Vosburgh E, Field M (1979) Coupled sodium-chloride influx across brush border of flounder intestine. J Membrane Biol 46:27–36
Frömter E (1972) The route of passive ion movement through the epithelium of Necturus gallbladder. J Membrane Biol 8:259–301
Fujimoto M, Kubota T (1976) Physiochemical properties of a liquid ion exchanger microelectrode and its applications to biological fluids. Jap J Physiol 26:631–650
Fujimoto M (1981) Intracellular ion activity measurements in renal tubular epithelium. In: Karger S (Hrsg) Proc 8th Int Congr Nephrol, Athens, pp 949–955
Geck P, Pietrzyk C, Burckhardt BC, Pfeiffer P, Heinz E (1980) Electrically silent cotransport of Na+, K+ and Cl− in Ehrlich cells. Biochim Biophys Acta 600:432–447
Gertz K (1962) Direct measurement of the transtubular flux of electrolytes and non-electrolytes in the intact rat kidney. Proc 22nd Int Congr Physiol Sciences, Leyden, p 370
Giebisch G (1979) Renal potassium transport. In: Giebisch G, Tosteson D, Ussing HH (Hrsg) Transport across biological membranes. Springer, Berlin Heidelberg New York
Greger R (1981) Chloride reabsorption in the rabbit cortical thick ascending limb of the loop of Henle — a sodium dependent process. Pflügers Arch 390:38–43
Greger R, Schlatter E (1981) Presence of luminal K+, a prerequisite for active NaCl transport in the thick ascending limb of Henle's loop of rabbit kidney. Pflügers Arch 392:92–94
Hansen LL, Schilling AR, Wiederholt M (1981) Effect of calcium, furosemide and chlorothiazide on net volume reabsorption and basolateral membrane potential of the distal tubule. Pflügers Arch 389:121–126
Imai M (1977) Effect of bumetanide and furosemide on the thick ascending limb of Henle's loop of rabbits and rats perfused in vitro. Europ J Pharm 41:409–416
Khuri RN, Wiederholt M, Strieder N, Giebisch G (1975) Effects of flow rate and potassium intake on distal tubular potassium transfer. Amer J Physiol 228:1249–1261
Machen TE, McLennan WL (1980) Na+-dependent H+ and Cl− transport in in vitro frog gastric mucosa. Amer J Physiol 238:G403-G413
McManus TJ, Schmidt WF III. (1978) Ion and co-ion transport in avian red cells. In: Hoffman JF (Hrsg) Membrane transport processes, Vol. 1. Raven Press, New York, pp 97–106
Nellans HN, Frizzell RA, Schultz SG (1973) Coupled sodium-chloride influx across the brush border of rabbit ileum. Amer J Physiol 225:467–475
Oberleithner H, Giebisch G (1981) Effects of furosemide and low chloride on potassium (K) transport across Amphiuma distal tubule — linkage of potassium (K) with chloride (Cl) reabsorption. 8th Int Congr of Nephrol Athens, 7–12 June, Astr TT-079
Oberleithner H, Giebisch G (1981) Mechanism of potassium transport across distal tubular epithelium of Amphiuma. In: MacKnight ADC, Leader JP (Hrsg) Epithelial ion and water Transport. Raven Press, New York, pp 97–105
Oberleithner H, Guggino W, Giebisch G (1981) The cellular mechanism of potassium adaptation in the distal amphibian nephron. Proc Physiol Soc 76P
Oberleithner H, Guggino W, Giebisch G (1982) Mechanism of distal tubular chloride transport in Amphiuma kidney. Amer J Physiol 242:F331-F339
Palfrey HC, Feit PW, Greengard P (1980) cAMP-stimulated cation cotransport in avian erythrocytes: inhibition by “loop” diuretics. Amer J Physiol 238:C139-C148
Pfaller W, Fischer WM, Strieder N, Wurnig H, Deetjen P (1974) Morphologic changes of cortical nephron cells in potassium-adapted rats. Laboratory Invest 31:678–684
Rocha AS, Kokko JP (1973) Sodium chloride and water transport in the medullary thick ascending limb of Henle: Evidence for active chloride transport. J Clin Invest 52:612–623
Schultz SG (1977) Sodium-coupled solute transport by small intestine: a status report. Amer J Physiol 233:E249-E254
Shindo T, Spring KR (1981) Chloride movement across the basolateral membrane of proximal tubule cells. J Membrane Biol 58:35–42
Silva P, Brown RS, Epstein FH (1977) Adaptation to potassium. Kidney Int 11:466–475
Stanton B, Biemesderfer D, Wade JB, Giebisch G (1981) Structural and functional study of the rat distal nephron: Effects of potassium adaptation and depletion. Kidney Int 19:36–48
Steiner RA, Oehme M, Ammann D, Simon W (1979) Neutral carrier sodium ion-selective microelectrode for intracellular studies. Analyt Chem 51:351–353
Stoner LC (1977) Isolated perfused amphibian renal tubules: the diluting segment. Amer J Physiol 233:F438-F444
Sullivan WJ (1968) Electrical potential differences across distal renal tubules of Amphiuma. Amer J Physiol 214:1096–1103
Thomas RC (1974) Intracellular pH of snail neurones measured with a new pH-sensitive glass microelectrode. J Physiol 238:159–180
Velázquez H, Wright FS, Good DW (in press) Luminal influences on potassium secretion: Chloride replacement with sulfate. (Accepted for publication by Amer J Physiol)
Walker JL (1971) Ionic specific liquid ion exchanger microelectrodes. Anal Chem 43:89A-93A
Wiederholt M, Sullivan WJ, Giebisch G (1971) Potassium and sodium transport across single distal tubules of Amphiuma. J Gen Physiol 57:495–525
Windhager EE, Giebisch G (1965) Electrophysiology of the nephron. Physiol Rev 45:214
Wright FS (1977) Sites and mechanisms of potassium transport along the renal tubule. Kidney Int 11:415–432
Wright FS, Strieder N, Fowler NB, Giebisch G (1971) Potassium secretion by distal tubule after potassium adaptation. Amer J Physiol 221:437–448
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Supported by NIH grant PHS AM 17433, by the Fogarty foundation (5 FO5 TWO 3865-02) and by Österreichischer Forschungsrat, Proj. No.: 4366
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Oberleithner, H., Giebisch, G., Lang, F. et al. Cellular mechanism of the furosemide sensitive transport system in the kidney. Klin Wochenschr 60, 1173–1179 (1982). https://doi.org/10.1007/BF01716719
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DOI: https://doi.org/10.1007/BF01716719