Transepithelial Ca2+ and Mg2+ transport in the cortical thick ascending limb of Henle's loop of the mouse is a voltage-dependent process.
The mechanisms responsible for transepithelial Ca2+ and Mg2+ in transport in the isolated perfused cortical thick ascending limb (cTAL) of Henle's loop of the mouse nephron were investigated by measuring transepithelial voltages (PDte) and transepithelial ion net fluxes (JNa, JCl, JK, JCa, JMg) by electron microprobe analysis. In the presence of furosemide (10(-4) mol.l-1, lumen) and diphenylamine-2-carboxylate (DPC, 10(-4) mol.l-1, bath), known inhibitors of NaCl reabsorption in the TAL, Ca2+ and Mg2+ reabsorption was completely inhibited. In the presence of furosemide, JCa fell from 0.75 +/- 0.07 to -0.08 +/- 0.09 pmol.min-1.mm-1 (n = 5), and JMg from 0.47 +/- 0.04 to -0.01 +/- 0.11 pmol.min-1.mm-1 (n = 5). In the presence of DPC, JCa fell from 0.57 +/- 0.08 to -0.07 +/- 0.11 pmol.min-1.mm-1 (n = 5), and JMg from 0.16 +/- 0.02 to -0.11 +/- 0.07 pmol.min-1.mm-1 (n = 5). With furosemide, inhibition of Ca2+ and Mg2+ transport was paralleled by a 93% inhibition of NaCl reabsorption, while in the presence of DPC there was a 60% reduction of NaCl reabsorption. These effects were fully reversed after removal of the inhibitors from the lumen or bath solutions. In the absence of active NaCl transport, a lumen-to-bath directed-NaCl gradient (lumen: 150 mM NaCl + furosemide, bath: 50 mM NaCl + 200 mM mannitol) generated a negative transepithelial dilution potential of -13.8 +/- 1.1 mV (n = 8) which induced a significant Ca2+ and Mg2+ secretion into the tubular lumen of -0.59 +/- 0.06 and -0.43 +/- 0.05 pmol.min-1.mm-1 (n = 8), respectively. A bath-to-lumen-directed NaCl gradient, on the other hand, (lumen: 50 mM NaCl + furosemide, bath: 150 mM NaCl) generated a positive transepithelial dilution potential of +15.9 +/- 0.6 mV (n = 7), inducing a significant Ca2+ and Mg2+ reabsorption of 0.62 +/- 0.08 and 0.38 +/- 0.07 pmol.min-1.mm-1 (n = 7), respectively. Linear regression analysis of individual Ca2+ and Mg2+ net flux data versus voltage indicated that JCa and JMg were highly correlated to PDte. In conclusion, these data indicate that transepithelial Ca2+ and Mg2+ reabsorption in the mouse cTAL is predominantly a passive process, driven by the lumen-positive PDte.
Di Stefano
A
A
Département de Biologie Cellulaire et Moléculaire, CE Saclay, Gif-sur-Yvette, France.
Roinel
N
N
de Rouffignac
C
C
Wittner
M
M
D000042:Absorption / Q000187:drug effects; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002118:Calcium / Q000378:metabolism*; D004594:Electrophysiology; D004848:Epithelium / Q000378:metabolism; D005260:Female; D005665:Furosemide / Q000494:pharmacology; D008138:Loop of Henle / Q000378:metabolism*; D008274:Magnesium / Q000378:metabolism*; D051379:Mice; D008815:Mice, Inbred Strains; D062367:ortho-Aminobenzoates / Q000494:pharmacology
D062367:ortho-Aminobenzoates; D005665:Furosemide; C001209:fenamic acid; D008274:Magnesium; D002118:Calcium