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  • 1
    ISSN: 1432-2013
    Schlagwort(e): ADH ; Mouse nephron ; Microperfusion ; NaCl transport ; Mg2+ transport ; Ca2+ transport ; Electron microprobe analysis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract The effect of antidiuretic hormone (ADH) on transepithelial Na+, Cl−, Ca2+ and Mg2+ net fluxes (JNa, JCl, JMg, JCa) was investigated in isolated perfused cortical thick ascending limb segments (cTAL) of the mouse nephron, using the microperfusion technique and the electron microprobe analysis to determine the ionic composition of the collected tubular fluid. Simultaneously, the transepithelial potential difference (PDte) and the transepithelial resistance (Rte) were recorded. Prior to the flux measurements cTAL segments were perfused for one hour. During this equilibration period PDte decreased significantly from +19.9±1.6 to +14.9±1.l mV and Rte increased from 30.6±3.5 Ωcm2 to 38.8±2.4 Ωcm2 (n=7), reflecting a decline in NaCl transport. After ADH was added to the bath solution at 10−10 mol.l−1, PDte increased from +14.4±1.1 to +18.0±1.5 mV, accompanied by a rise in JNa and JCl from 205±11 to 273±19 and from 216±12 to 283±21 pmol.min−1.mm−1 (n=7), respectively. JCa and JMg also increased from 0.81±0.07 to 1.50±0.12 and from 0.43±0.11 to 0.76±0.08 pmol.min−1.mm−1 (n=7), respectively. All these effects were fully reversible after withdrawal of the hormone. In conclusion our data indicate that ADH stimulates divalent cation transport and NaCl transport in the cortical thick ascending limb of Henle's loop of the mouse.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1432-2013
    Schlagwort(e): Transepithelial NaCl net fluxes ; Na+/H+ exchanger ; Cl−/HCO 3 − exchanger ; Isolated perfused tubule ; Electron microprobe analysis ; Cortical thick ascending limb of Henle's loop
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract The aim of the present study was to investigate whether bicarbonate buffer (CO2 + HCO 3 − ) is required to sustain maximal NaCl transport in the cortical thick ascending limb of Henle's loop (cTAL) of the mouse. Transepithelial Na+ and Cl− net fluxes (J Na, J Cl, pmol min−1 mm−1), measured by electron microprobe analysis, were similar irrespective of the presence or absence of CO2 + HCO 3 − in luminal and bathing solutions J NaCl with CO2 + HCO 3 − =203±25 pmol min−1 mm−1; J NaCl without CO2 + HCO 3 − =213±13 pmol min−1 mm−1, n=14). Furthermore the transepithelial potential difference, V te, the transepithelial resistance, R te, and the basolateral membrane potential, V bl, were unaffected by CO2 + HCO 3 − . In the absence of CO2 + HCO 3 − , V te was +17.0±1.7 mV(n=9) (lumen positive), R te was 28±2 Ω cm2 (n=9) and V bl was −76±4 mV (n=6). In the presence of CO2 + HCO 3 − , V te, R te and V bl were +15.9±1.5 mV, 29±1 Ω cm2 and −73±5 mV, respectively. 4-Acetamido-4′-isothiocyanatostilbene-2,2′-disulphonic acid (SITS; 0.1 mmol l−1) and amiloride (1 mmol l−1) added to the (CO2 + HCO 3 − )-containing lumen perfusate were without effect on V te and R te. Finally, the effect of furosemide (0.1 mmol l−1) on V te and V bl in the presence of CO2 + HCO 3 − was investigated. Furosemide reversibly decreased V te from +13.7±1.1 mV to +1.7±0.7 mV (n=6) and hyperpolarized Vbl from −70±1 to −89±3 mV (n=5), suggesting passive distribution of Cl− across the basolateral membrane. In conclusion, these data suggest that active NaCl transport in the cTAL of the mouse does not require the presence of CO2 + HCO 3 − .
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    ISSN: 1432-2013
    Schlagwort(e): Ca2+ transport ; Mg2+ transport ; Electron microprobe analysis ; Cortical thick ascending limb ; Furosemide ; Parathyroid hormone ; Paracellular shunt pathway permeability ; Tight junctions
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Recent studies from our laboratory have shown that in the cortical thick ascending limb of Henle's loop of the mouse (cTAL) Ca2+ and Mg2+ are reabsorbed passively, via the paracellular shunt pathway. In the present study, cellular mechanisms responsible for the hormone-stimulated Ca2+ and Mg2+ transport were investigated. Transepithelial voltages (PDte) and transepithelial ion net fluxes (J Na, J Cl, J K, J Ca, J Mg) were measured in isolated perfused mouse cTAL segments. Whether parathyroid hormone (PTH) is able to stimulate Ca2+ and Mg2+ reabsorption when active NaCl reabsorption, and thus PDte, is abolished by luminal furosemide was first tested. With symmetrical lumen and bath Ringer's solutions, no Ca2+ and Mg2+ net transport was detectable, either in the absence or in the presence of PTH. In the presence of luminal furosemide and a chemically imposed lumen-to-bath directed NaCl gradient, which generates a lumen-negative PDte, PTH slightly but significantly increased Ca2+ and Mg2+ net secretion. In the presence of luminal furosemide and a chemically imposed bath-to-lumen-directed NaCl gradient, which generates a lumen-positive PDte, PTH slightly but significantly increased Ca2+ and Mg2+ net reabsorption. In view of the observed small effect of PTH on passive Ca2+ and Mg2+ movement, a possible interference of furosemide with the hormonal response was considered. To investigate this possibility, Ca2+ and Mg2+ transport was first stimulated with PTH in tubules under control conditions. Then active NaCl reabsorption was abolished by furosemide and the effect of PTH on J Ca and J Mg measured. In the absence of PDte and under symmetrical conditions, no Ca2+ and Mg2+ transport was detectable, either in the presence or absence of PTH. In the presence of a bath-to-lumen-directed NaCl gradient, Ca2+ and Mg2+ reabsorption was significantly higher in the presence than in the absence of PTH. Finally, when active NaCl transport was not inhibited by furosemide, but reduced by a bath-to-lumen-directed NaCl gradient, PTH strongly increased J Ca and J Mg, whereas only a small increase in PDte was noted. In conclusion, these data suggest that PTH exerts a dual action on Ca2+ and Mg2+ transport in the mouse cTAL by increasing the transepithelial driving force for Ca2+ and Mg2+ reabsorption through hormone-mediated PDte alterations and by modifying the passive permeability for Ca2+ and Mg2+ of the epithelium, very probably at the level of the paracellular shunt pathway.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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