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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular medicine 61 (1983), S. 1029-1037 
    ISSN: 1432-1440
    Keywords: Intracellular Na+, K+, Cl−, H+ HCO 3 − , Ca2+ ; Ouabain ; Phenylalanine ; Glucose ; Renal tubular transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary To disclose possible regulatory mechanisms, the potential difference across the peritubular cell membrane (PDpt) and intracellular activities of sodium (Na i + ), potassium (K i + ), calcium (Ca i 2+ ), bicarbonate (HCO 3i − ) and chloride (Cl i − ) have been traced continuously during inhibition of Na+/K+-ATPase with ouabain. Within 31±4 min following application of ouabain, PDpt decreases (from 57±2 mV) to half and K i + by 37.7±2.2 mmol/l (from 63.5±1.9 mmol/l), Na i + increases by 35.1±4.1 mmol/l (from 13.2±2.4 mmol/l), Ca i 2+ by 0.17 ± 0.2 µmol/l (from 0.09 µmol/l), HCO 3i − by 3.0±1.1 mmol/l (from 15.3±2.0 mmol/l) and Cl i − by 6.2±1.0 mmol/l (from 14.4±1.6 mmol/l). Within the same time the luminal and peritubular cell membrane resistances increase 45±15% and 53±17%, respectively. The increase of the resistances is mainly due to a decrease of K+ conductance, which in turn mainly accounts for the depolarisation of PDpt. Additional experiments demonstrate that the K+ conductance of the peritubular cell membrane is sensitive to the cell membrane potential difference and possibly linked to Na+/K+-ATPase activity. The decline of PDpt probably accounts for intracellular alkalinisation which in turn reduces Na+/H+ exchange. Na+-coupled transport of glucose and phenylalanine decrease in linear proportion to PDpt. The transport of these and probably of similar substances represents the main threat to electrolyte homeostasis of the cells.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Ouabain ; Cell membrane potential ; Cell membrane resistance ; Potassium conductance ; Bicarbonate conductance ; Proximal tubule ; Amphibian kidney
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract According to a previous study from this laboratory, the electrochemical gradient for potassium across the peritubular cell membrane of proximal tubules in the isolated perfused frog kidney increases following the application of ouabain. In order to test, if this phenomenon were due to a decrease of potassium conductance, the effects of ouabain on cell membrane resistances and the sensitivity of the peritubular cell membrane potential difference (PDpt) to step changes of peritubular potassium and bicarbonate concentration were studied. In the absence of ouabain, PDpt averaged −60±3 mV (n=25). A step increase of peritubular potassium concentration from 3 to 18 mmol/l (pH 8.07) depolarises PDpt (ΔPDk) by +24±2 mV (n=8). An increase of bicarbonate from 20 to 40 mmol/l (pH 8.07) hyperpolarises PDpt (ΔPDb) by −2.8±0.4 mV (n=9). The resistance of the luminal and peritubular cell membranes in parallel (R m) amounts to 45±9 kΩ cm (tubule length) (n=4) and the voltage divider ratio (VDR) to 1.4±0.2 (n=7). The resistance of the cellular cable (cellular core,R c) approaches 131±37 MΩ/cm (n=4). Peritubular application of 0.1 mmol/l ouabain leads to a gradual decline of PDpt (t 1/2 approx. 30 min), to an increase ofR m, a decrease of ΔPDk and an increase of ΔPDb. VDR andR c are not changed significantly. The data point to a functional link between the sodium/potassium ATPase and the potassium conductance of the peritubular cell membrane.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2013
    Keywords: Frog kidney ; Proximal tubule ; Glucose transport ; Ouabain ; Cell membrane potential ; Intracellular sodium ; Microelectrodes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Inhibition of basolateral Na+/K+ ATPase by ouabain eventually abolishes transport of glucose. The present study was performed to test, if this effect is due to a dissipation of the electrochemical gradient for sodium or due to a regulatory inhibition of sodium-coupled glucose entry across the luminal membrane at increasing intracellular sodium activity. To this end, proximal convoluted tubules of the doubly perfused isolated frog kidney were perfused alternatively with solutions containing either 5 mmol/l glucose or raffinose. The potential difference across the peritubular cell membrane (PDpt) and across the epithelium (PDpt) has been recorded with conventional and across the peritubular cell membrane with ion selective microelectrodes (PDpt). In the absence of luminal glucose PDpt is (±SEM) −54.0±2.4 mV, PDte=−1.2±2.0 mV and PD pt Na =−96±5 mV. The electrochemical gradient for sodium (μNa+) amounts to 95 mV and intracellular sodium activity to 14 mmol/l (extracellular sodium activity is 74 mmol/l). Luminal application of glucose leads to a rapid depolarisation of PDpt (ΔPDpt=8.6±0.9 mV and PD pt Na (ΔPD pt Na =11.1±3.0 mV) and to hyperpolarisation of PDte (ΔPDte=−0.8±0.2 mV). The peritubular application of ouabain leads to a gradual, reversible and proportional decline of PDpt, PD pt Na and μNa+. Glucose induced ΔPDpt and ΔPD pt Na decrease in parallel to PDpt and PD pt Na , resp. In a separate series, the lumped conductance (G m) of the luminal and basolateral cell membrane has been determined, which amounts to 2.4±0.3 μS/mm (tubule length).G m decreases 23±4%, when PDpt is decreased to half. ΔPDpt andG m allow the calculation of an apparent transport rate (T Glu). Following the application of ouabain,T Glu decreases in linear proportion to PDpt and PD pt Na . There is no evidence for a significant regulatory inhibition ofT Glu. Rather, glucose transport operates in linear proportion to the potential difference across the luminal membrane.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2013
    Keywords: Ouabain ; Frog kidney ; Proximal tubule ; Ion sensitive microelectrodes ; Na+/K+ pump ; Intracellular K+, Na+, Cl−, pH, Ca2+
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using conventional and ion selective microelectrodes, the effect of ouabain (10−4 mol/l) on peritubular cell membrane potential (PDpt), on intracellular pH (pHi) as well as on the intracellular ion activities of Cl− (Cl i − ), K+ (K i + ), Na+ (Na i + ) and Ca2+ (Ca i 2+ ) was studied in proximal tubules of the isolated perfused frog kidney. In the absence of ouabain (PDpt=−57.0±1.9 mV), the electrochemical potential difference of chloride (apparent {ie6-1} and of potassium {ie6-2} is directed from cell to bath, of H+ {ie6-3}, of Na+ {ie6-4} and of Ca2+ {ie6-5} from bath to cell. Ouabain leads to a gradual decline of PDpt, which is reduced to half (PDpt, 1/2) within 31±4.6 min (in presence of luminal glucose and phenylalanine), and to a decline of the absolute values of apparent {ie6-6}, of {ie6-7}, {ie6-8} and {ie6-9}. In contrast, an increase of {ei6-10} is observed. At PDpt, 1/2 apparent Cl i − increases by 6.2±1.0 mmol/l, pHi by 0.13±0.03, Ca i 2+ by 185±21 nmol/l, and Na i + by 34.2±4.6 mmol/l, whereas K i + decreases by 37.7±2.2 mmol/l. The results suggest that the application of ouabain is followed by a decrease of peritubular cell membrane permeability to K+, by an accumulation of Ca2+, Na+ and HCO 3 - in the cell and by a dissipation of the electrochemical Cl− gradient.
    Type of Medium: Electronic Resource
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