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  • 1
    ISSN: 1432-2013
    Schlagwort(e): Distal tubule ; Sodium chloride transport ; Cellular sodium activity ; Furosemide ; Amphiuma kidney
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Previous experiments had demonstrated that cell chloride activities in early distal tubule cells of Amphiuma are above equilibrium distribution. Chloride activities fell sharply towards electrochemical equilibrium following perfusion of the tubular lumen with furosemide or with sodium-free solutions. These results suggested a furosemide-sensitive sodium chloride cotransport system in the luminal cell membrane. The present experiments were carried out to evaluate directly the electrochemical driving forces acting on sodium ions under similar experimental conditions. Intracellular sodium activity measurements were performed in the doublyperfused kidney of Amphiuma by means of single-barreled liquid ion-exchange microelectrodes. Basolateral cell membrane potential and resistance ratio measurements of tubular cell membranes were also carried out under control conditions and after inhibition of chloride transport by luminal application of furosemide (5 · 10−5 mol/l) or by omission of chloride. Control conditions were characterized by a steep downhill electrochemical gradient for sodium ions from lumen to cell. Inhibition of chloride transport led to a sharp decrease of intracellular sodium activity and to hyperpolarization of the peritubular membrane potential while the resistance ratio of the tubular cell membranes did not change significantly. These results demonstrate the presence of low cellular sodium activities in early distal tubule cells. The sharp decline of cell sodium after furosemide and after luminal chloride removal is consistent with inhibition of a sodium chloride cotransport system and continued peritubular sodium extrusion. The latter can increase the electrochemical gradient of sodium ions beyond that observed under control conditions.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1432-2013
    Schlagwort(e): Potassium ; Microelectrodes ; Potassium transport ; Furosemide ; Potassium adaptation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Previous experiments in the early distal tubule of the doubly perfused kidney ofAmphiuma demonstrated net reabsorption of potassium (K) which is reversed to net K secretion after K adaptation. Furthermore, it is known that this particular segment exhibits extensive chloride (Cl) net reabsorption which depends on the presence of sodium (Na) and which is inhibited by furosemide. In order to test for a possible interrelationship between NaCl and K transport, K activity in lumen and cell, transepithelial electrical potential difference, peritubular cell membrane potentials and volume reabsorption were measured in control animals and after K adaptation, in presence and absence of furosemide. In control animals the direction of net K transport is reversed from reabsorption to secretion upon addition of furosemide or following the removal of Cl from the tubular lumen. Volume reabsorption is inhibited by some 80%. In K adapted animals a similar inhibition of volume reabsorption is observed, however K secretion is not further enhanced. In control as well as in K-adpated animals intracellular K activities are still above electrochemical equilibrium after furosemide. The data suggest that a common transport system for Na, Cl and K is present in the luminal cell membrane which is inhibited by furosemide, K secretion observed in controls after furosemide and in K-adapted animals is driven by the cell to lumen electrochemical gradient for K across the K permeable luminal cell membrane. The shift of the luminal pump-leak system towards K secretion following K adaptation may be explained by an increase of the luminal K conductance and/or by a reduction of the activity of the luminal cotransport system. However, other mechanisms may also contribute to the observed phenomenon of K adaptation and cannot be ruled out at present.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    Digitale Medien
    Digitale Medien
    Springer
    Journal of molecular medicine 60 (1982), S. 1173-1179 
    ISSN: 1432-1440
    Schlagwort(e): Distal tubule ; Furosemide ; Ion-sensitive microelectrodes ; Sodium chloride cotransport ; Potassium adaptation ; Distaler Tubulus ; Furosemid ; Ionen-sensitive Microelektroden ; Natrium Chlorid Kotransport ; Kaliumadaptation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Beschreibung / Inhaltsverzeichnis: 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.
    Notizen: 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.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 4
    ISSN: 1432-2013
    Schlagwort(e): Amphibian kidney ; Potassium ; Chloride ; Barium ; Furosemide
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Experiments in the amphibian early distal tubule have shown that Cl transport is secondarily active, coupled to the flux of Na and dependent on the presence of luminal K. Omission of luminal K results in a decrease of cellular Na, a finding that suggests inhibition of luminal Na entry. In the present study intracellular chloride activity (Cli) and peritubular cell membrane potentials (PDpt) were evaluated before and after omission of luminal K. Furthermore, the effect of inhibition of the luminal K conductance by barium on the electrochemical gradient of Cl (E te Cl ) across the distal epithelium was determined at static head conditions. Experiments were performed in early distal segments of the isolated perfused kidney ofAmphiuma andRana esculenta. Cli and PDpt were measured simultaneously in single cells by double barreled Cl sensitive microelectrodes in the presence and absence of luminal K. E te Cl was determined at zero net flux conditions with single barreled electrodes in control tubules, in the presence of barium (3·10−3 mol/l) and in the presence of furosemide (5·10−5 mol/l). In 26 individual cellular impalements omission of luminal K hyperpolarized PDpt from 72.5±1.2 to 90.0±1.9 mV (cell interior negative). Concomitantly, Cli fell from 8.5±0.4 to 5.4±0.3 mmol/l. Both effects occurred within seconds and were fully reversible. Addition of barium to the luminal fluid diminished E te Cl (directed lumen positive) from a control value of 39.5±1.4 mV to 28.5±2.5mV. E te Cl could be further diminished to 14.1±2.1 mV and to 1.3±0.5 mV after application of barium on both sides and after luminal application of furosemide, respectively. The experiments indicate that active Cl uptake across the luminal cell membrane depends critically on the presence of luminal K. Omission of luminal K achieved either by perfusing the lumen with K-free solutions or by inhibition of K back flux from the cell interior into the lumen by barium reduces Cl reabsorption. Together with previous data on the K dependence of the Na uptake the present experiments support the hypothesis of a common transport system for K, Na, Cl located in the luminal cell membrane.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 5
    ISSN: 1432-2013
    Schlagwort(e): Diluting segment ; Furosemide ; K+-conductance ; Amiloride
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Experiments were performed in the isolated perfused kidney of K+ adaptedRana pipiens to investigate the relationship between luminal K+ conductance and H+ transport in cells of the diluting segment. Inhibition of luminal Na+/H+ exchange by amiloride or by omission of luminal Na+ blocked luminal K+ conductance. Acidification of the kidney perfusate by elevation of pCO2 also reduced luminal K+ conductance. This effect could be prevented by furosemide. Since the steepest transcellular Na+ potential difference, directed from the lumen into the cell, is found when luminal Na+/Cl−/K+ cotransport is inhibited by furosemide, we conclude that luminal Na+/H+ exchange is most efficient at these conditions and thus could attenuate intracellular acidification.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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