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  • 1
    ISSN: 1432-2013
    Keywords: Ca2+ transport ; Mg2+ transport ; Electron microprobe analysis ; Cortical thick ascending limb ; Furosemide ; Parathyroid hormone ; Paracellular shunt pathway permeability ; Tight junctions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: 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.
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  • 2
    ISSN: 1432-2013
    Keywords: ADH ; Mouse nephron ; Microperfusion ; NaCl transport ; Mg2+ transport ; Ca2+ transport ; Electron microprobe analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: 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.
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  • 3
    ISSN: 1432-2013
    Keywords: Renal Tubule ; Phosphate Transport ; Sodium Dependence ; Micropuncture ; Microperfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The standing droplet method has been used in combination with the peritubular perfusion of blood capillaries to determine the build up of transtubular concentration differences of phosphate (P i ) in the renal proximal convoluted tubule of parathyroidectomized rats. Electron probe analysis was used to estimate P i . At zero time both the intraluminal and the contraluminal P i concentration was 2 mM. The time dependent decrease of the intraluminal P i concentration was approximately 4 times faster in the early than in the late proximal convoluted tubule. After 45 sec an intraluminal steady state concentration of 0.20 mM P i was achieved in the early part. In the late part the intraluminal P i concentration approached a steady state value of 0.54 mM at 120 sec. When sodium free solutions were used the intraluminal P i concentration increased to 2.22 mM in the earlier and to 2.76 mM in the late part. The data indicate that in the proximal convoluted tubule 1. The rate of phosphate reabsorption is greater in the early part than in the later part, and 2. phosphate reabsorption might occur as co-transport with Na+ ions.
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  • 4
    ISSN: 1432-2013
    Keywords: Extracellular fluid volume expansion ; Mg, Ca and Pi renal handling ; Electron microprobe analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: 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.
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  • 5
    ISSN: 1432-2013
    Keywords: Calcitonin ; Distal tubule ; Micropuncture ; Electron-probe ; Brattleboro rats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of human calcitonin (HCT) on the distal tubule were investigated by micropuncture in hormone-deprived rats, i.e. in the absence of parathyroid hormone, antidiuretic hormone and glucagon, which might have masked these effects. Two groups of rats were studied: hormone-deprived and hormone-deprived+HCT, infused at 1.0 mU/min\100 g b.w. In the urine, HCT markedly reduced Ca and Mg excretion whereas excretion of water, Na and K was not significantly affected. Along the distal tubule, HCT strongly enhanced Na, Cl, Mg, Ca and total solute reabsorption, decreased K secretion but did not alter water or phosphate transport. It is concluded that HCT stimulated Na, Cl, Ca and Mg reabsorption. If, as suggested, HCT also stimulated the reabsorptive component of K transport, the hormone should therefore elicit the same physiological effects in the distal tubule and the thick ascending limb.
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  • 6
    ISSN: 1432-2013
    Keywords: Micropuncture ; Dog ; Mg, Ca, Na, K, Pi reabsorption ; Electron probe analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Micropuncture and clearance experiments were done on normal dogs to investigate magnesium handling by proximal and distal nephron segments. Tubular fluid electrolytes were analyzed with the electron microprobe. Tubular fluid to ultrafilterable magnesium ratio (TF/UF magnesium) was observed to rise above unity but less than the TF/P inulin ratio generated along the proximal tubule. This is in contrast to the other major cations, the ratios of which remain close to unity as water is abstracted. Tubular fluid obtained from the distal tubule contained less magnesium than the glomerular filtrate (mean TF/UF magnesium of 0.6) indicating the loop of Henle is the major nephron segment reclaiming a significant portion of the filtered load. The faction of filtered load remaining at the distal sampling site was similar to the fraction appearing in the urine (8% vs 7%) indicating very little reabsorption beyond the distal tubule in these normal states.
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  • 7
    ISSN: 1432-2013
    Keywords: Micropuncture ; Mg, Ca and P Excretion ; Concentrating Mechanism ; Loop of Henle ; Electron Probe Analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cortical and papillary micropuncture experiments were carried out on Psammomys undergoing mild hypertonic salt diuresis. Tubular fluid was collected along the proximal tubule or at the early distal tubule level, and at the tip of the longest Henle loop.3H-inulin, Na, K, Cl, Mg, Ca and P concentrations, as well as osmotic pressure, were determined in all samples. The results indicate a) no large net water movement along the loop; b) substantial addition of Na, K, Cl, Mg, and to a lesser extent Ca, along the descending limb, in proportion to the gradient; c) the tubular flow rate of phosphorus remains constant at the tip of the loop irrespective of the gradient. The constancy of the load of Na, K, Cl, Mg and Ca delivered to the distal superficial nephron, irrespective of the urinary osmotic pressure, indicates that medullary recycling between the ascending and descending limbs exists for Mg, Cl, and Ca, and confirms its existence for Na and K. In contrast, phosphorus behaves like inulin along the descending limb. A general conclusion is that in Psammomys the concentrating process along the descending limb of Henle results mainly from net addition of solutes, and not from water withdrawal.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-2013
    Keywords: Brattleboro ; Antidiuretic hormone ; Calcitonin ; Parathyroid hormone ; Glucagon ; Electrolyte ; Micropuncture ; Electron probe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effects of (1-desamino-8-d-arginine) vasopressin (dDAVP) on water and electrolyte transport in the distal tubule were investigated by micropuncture. Since, in addition to antidiuretic hormone, parathyroid hormone, calcitonin and glucagon stimulate the adenylate-cyclase system in this nephron segment, experiments were performed on hormone-deprived rats, i.e. homozygous DI Brattleboro rats with reduced levels of endogenous parathyroid hormone, calcitonin and glucagon. Along the distal tubule, dDAVP enhanced water, Cl, Na and Ca reabsorption and sharply increased net K secretion. Phosphate transport was left unchanged and Mg reabsorption was not significantly altered by dDAVP between the early and late distal tubule. Antidiuretic hormone also slightly increased water filtration rate in the superficial nephron, which rose in proportion to whole kidney glomerular filtration rate. It is concluded that, in rats: 1) antidiuretic hormone stimulates water, NaCl and Ca absorption and enhances K secretion along the distal tubule and 2) the tubular effects of dDAVP on electrolyte transport in the loop and distal tubule are responsible for decreasing Mg and Ca urinary excretion.
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