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  • 1
    ISSN: 1432-2013
    Schlagwort(e): Extracellular fluid volume expansion ; Mg, Ca and Pi renal handling ; Electron microprobe analysis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Renal tubular handling of P, Ca, Mg and Na was studied in the rat both before and during mild hypertonic NaCl loading (ECVE), using micropuncture and clearance techniques and electron microprobe analysis. Micropuncture was performed at the late proximal and early distal tubule sites. ECVE significantly increased the urinary output of all four elements. In the case of Mg, the increase was relatively small and dependend of reabsorption all along the entire length of the nephron. For Ca, it depended on the inhibition of proximal reabsorption, partially compensated by increased reabsorption along the loop. For P, it depended on proximal inhibition, no important net phosphate movement occuring in the loop during both periods. Ca reabsorption was highly correlated to that of sodium along the proximal tubule and Henle's loop. This was not the case for Mg and P. In the loop, Ca and Mg reabsorption were closely related to the load delivered at the beginning of the structure. These observations are compatible with the view that tubular reabsorption of Ca and Mg is concentration rather than Tm limited, and that reabsorption of Ca, unlike that of Mg, is linked to the movements of sodium. Following ECVE, the difference between early distal and urinary deliveries increased significantly for Ca and P, but not for Mg. For phosphate, this difference accounted for by 45% of the delivery at the early distal tubule site, at variance with microinjection data obtained in the rat under similar salt loading conditions, which indicated that 17% only of the phosphate distal delivery were reabsorbed along the terminal segments. This discrepancy is discussed in terms of nephron functional heterogeneity.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    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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  • 3
    ISSN: 1432-2013
    Schlagwort(e): Kidney Tubules ; Single Nephron Filtration Rate ; Distribution of Single Nephron Filtration Rate ; Structure and Function
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Beschreibung / Inhaltsverzeichnis: Résumé Nous présentons dans ce travail une modification de la technique de Hanssen qui permet de la rendre quantitative. Un indicateur glomérulaire marqué (ferrocyanure de sodium14C) est perfusé à des rats anesthésiés jusqu'à l'obtention d'une concentration plasmatique stable. Puis 20 μl d'une solution à 10% de ferrocyanure non marqué sont injectés rapidement au dessus de l'artère rénale gauche. Quelques secondes plus tard, le pédicule rénal gauche est lié. Le ferrocyanure contenu dans le rein est alors précipité sous forme de bleu de Prusse insoluble, puis les tubules proximaux sont disséqués après macération. De la radioactivité contenue dans le néphron entre le glomérule et le précipité de ferrocyanure non marqué (radioactivité qui correspond donc au14C ferrocyanure filtré pendant un temps déterminé) et de la concentration plasmatique de cet indicateur, on déduit le débit de filtration (fg) du néphron considéré. Les résultats obtenus chez trois rats non diurétiques sont donnés à titre d'exemple. La fg de néphrons superficiels et profonds ayant une longueur moyenne de tubule proximal de 7,6 mm et 8,9 mm respectivement a été calculée à partir de l'équation de la droite de régression décrivant la variation de la fg en fonction de la longueur du tubule proximal: elle a été trouvée égale à 29,1 nl/min pour les néphrons superficiels et 40,1 nl/min pour les néphrons profonds. La validité de la technique est attestée 1. par le fait que la filtration glomérulaire totale du rein, calculée à partir des fg, est sensiblement identique à celle mesurée quelques minutes plus tôt (C in 2. par le fait que les valeurs des fg ainsi que les corrélations anatomo-fonctionnelles rapportées sont identiques à celles obtenues par la technique de Hanssen, 3. enfin par le fait que les fg des néphrons superficiels sont semblables à celles habituellement mesurées par microponction. Notre technique conserve tous les avantages de la méthode de Hanssen, mais elle en supprime les inconvénients liés à l'hétérogénéité de distribution dans le rein de l'indicateur marqué puisqu'ici, au lieu d'être administré en une seule injection, le ferrocyanure marqué est perfusé à l'animal; de plus, elle permet une mesure quantitative des valeurs individuelles de fg même sur des reins anuriques.
    Notizen: Summary We describe a modification of Hanssen's technique. Anesthetized rats were infused with a14C−Na-ferrocyanide solution. At equilibrium, 20 μl of a 10% non labelled ferrocyanide solution were injected as a short pulse above the left renal artery. A few seconds later, the left renal pedicle was tied. Ferrocyanide contained within the kidney was precipitated as insoluble Prussian blue and the proximal tubules were microdissected after maceration. From the radioactivity contained within the nephron from the glomerulus to the precipitate of non labelled ferrocyanide (radioactivity which is due to the14C filtered during a given time) and from the14C plasmatic concentration, we deduced the single glomerular filtration rate (SGFR) of the nephron. Three non diuretic rats are given as example. The SGFR of superficial and juxtamedullary nephrons was respectively equal to 29,1 nl/min and 40,1 nl/min when calculated for the mean value of the length of their proximal tubules (respectively 7.6 mm and 8.9 mm) from the equation of the regression line which described the correlation between SGFR and the length of the proximal tubule. The reliability of the technique is supported 1. by the fact that the value of the GFR of the whole kidney calculated from SGFR's was the same asC in measured just prior, 2. by the fact that SGFR values and the anatomo-physiological correlations were identical to those obtained with the Hanssen's technique, 3. and by the fact that the SGFR of superficial nephrons agree with most of those obtained with the micropuncture techniques. Most of the criticisms of Hanssen's technique, which arise from the observation that the bolus of labelled precipitate is sometimes inhomogenously distributed from one region of the kidney to another, are not valid here because instead of being injected as a pulse, the14C-ferrocyanide is infused continuously into the animal. Moreover, our technique allows measurements of SGFR's even in non urine forming kidneys.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 4
    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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  • 5
    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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  • 6
    Digitale Medien
    Digitale Medien
    Springer
    Pflügers Archiv 308 (1969), S. 260-276 
    ISSN: 1432-2013
    Schlagwort(e): Kidney Tubules ; Single Nephron Filtration Rates ; Heterogeneity of Nephron Function ; Structure and Function ; Nierentubuli ; Einzelnephronfiltrationsrate ; Unterschiede der Nephronfunktion ; Struktur und Funktion
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Non-diuretic rats and saline diuretic psammomys received 0.05 ml14C Na ferrocyanide i.v. Their kidneys were frozen in vivo 8–10 sec later. The quantity of14C ferrocyanide precipitated as prussian blue (Hanssen) in the lumen of a microdissected proximal tubule was proportional to its glomerular filtration rate (gfr). Distance between dye front and glomerulus was proportional to mean flow velocity (v). Fractional water reabsorption at the dye front was calculated using gfr, v and luminal radius. There were large differences in gfr, v and fractional water reabsorption between superficial and deep nephrons. Published micropuncture data agrees quantitatively with our results. Glomerular volume, tubular length and luminal radius increased from superficial to juxtamedullary proximal tubules in both species. Filtration rate varied directly with glomerular volume suggesting an influence of capillary surface area on filtration. Tubular length and luminal radius correlated with gfr. Proximal water reabsorption rate correlated with luminal radius in psammomys but not in rat tubules. Water reabsorption by the entire proximal tubule decreased with length in rat tubules but increased in psammomys. The discrepancy may be due to differences in blood flow distribution which alter gfr and/or water reabsorption in superficial relative to deep nephrons.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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