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  • 1
    ISSN: 1432-2013
    Keywords: ADH ; Transepithelial ion net fluxes ; Na+, Cl−, K+, Ca2+ and Mg2+ transport ; Electron microprobe ; Mouse kidney ; Cortical and medullary thick ascending limb of Henle's loop ; In vitro microperfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effect of antidiuretic hormone (arginine vasopressin, AVP) on transepithelial Na+, Cl−, K+, Ca2+ and Mg2+ net transports was investigated in medullary (mTAL) and cortical (cTAL) segments of the thick ascending limb (TAL) of mouse nephron, perfused in vitro. Transepithelial net fluxes (J Na +,J Cl −,J K +,J Ca 2+,J Mg 2+) were determined by electron probe analysis of the collected tubular fluid. Transepithelial potential difference (PDte) and transepithelial resistance (Rte) were measured simultaneously. cTAL segments were bathed and perfused with isoosmolal, HCO 3 − containing Ringer solutions, mTAL segments were bathed and perfused with isoosmolal HCO 3 − free Ringer solutions. In cTAL segments, AVP (10−10 mol·l−1) significantly increasedJ Mg 2+ andJ Ca 2+ from 0.39±0.08 to 0.58±0.10 and from 0.86±0.13 to 1.19±0.15 pmol·min−1 mm−1 respectively. NeitherJ Na + norJ Cl −, (J Na +: 213±30 versus 221±28 pmol·min−1 mm−1,J Cl −: 206±30 versus 220±23 pmol·min−1 mm−1) nor PDte (13.4±1.3 mV versus 14.1±1.9 mV) or Rte (24.6±6.5Ω cm2 versus 22.6±6.4Ω cm2) were significantly changed by AVP. No significant effect of AVP on net K+ transport was observed. In mTAL segments, Mg2+ and Ca2+ net transports were close to zero and AVP (10−10 mol·l−1) elicited no effect. However NaCl net reabsorption was significantly stimulated by the hormone,J Na + increased from 107±33 to 148±30 andJ Cl − from 121±33 to 165±32 pmol·min−1 mm−1. The rise inJ NaCl was accompanied by an increase in PDte from 9.0±0.7 to 13.5±0.9 mV and a decrease in Rte from 14.4±2.0 to 11.2±1.7 Ω cm2. No K+ net transport was detected, either under control conditions or in the presence of AVP. To test for a possible effect of HCO 3 − on transepithelial ion fluxes, mTAL segments were bathed and perfused with HCO 3 − containing Ringer solutions. With the exception ofJ Ca 2+ which was significantly different from zero (J Ca 2+: 0.26±0.06 pmol·min−1 mm−1), net transepithelial fluxes of Na+, Cl−, K+ and Mg2+ were unaffected by HCO 3 − . In the presence of AVP,J Mg 2+ andJ Ca 2+ were unaltered whereasJ NaCl was stimulated to the same extent as observed in the absence of HCO 3 − . In conclusion our results indicate heterogeneity of response to AVP in cortical and medullary segments of the TAL segment, since AVP stimulates Ca2+ and Mg2+ reabsorption in the cortical part and Na+ and Cl− reabsorption in the medullary part of this nephron segment.
    Type of Medium: Electronic Resource
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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: 1432-1424
    Keywords: Water fluxes ; Na fluxes ; proximal tubule microperfusion ; Li substitution ; rat kidney
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The relationship between water and sodium movements through the mammalian proximal convoluted tubule was investigated by substituting lithium for sodium. Proximal convoluted rat Kidney tubules were perfusedin vivo with a Ringer solution containing 107 meq/liter lithium and 42 meq/liter sodium. Several micropunctures were made along the same nephron, and [3H] inulin, [14C] glucose,22Na, osmolality, Na, Mg and Cl were determined on each sample. Measurements of22Na showed that sodium and lithium diffusion rates were practically identical throughout the entire epithelium. A one- for-one exchange of sodium for lithium induced a negative trans-epithelial net flux of Na from plasma to lumen. However, despite this negative flux, a positive net water movement was measured from lumen to plasma. This movement was proportional both to glucose reabsorption and to the rise in the chloride concentration, two mechanisms known to be dependent on the trans-cellular movement of sodium. It was therefore concluded that the net water flux was a function of the unidirectional transcellular net flux of Na. Rabbit proximal convoluted tubules were perfusedin vitro with a solution containing 75 meq/liter Li and 75 meq/liter Na on both the luminal and peritubular sides. Under these conditions, the water reabsorption rate dropped to half its control value. Water movement was therefore a function of the external sodium concentration, which in turn probably regulates the intracellular Na concentration.
    Type of Medium: Electronic Resource
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