Evidence supporting a role for KCl cotransporter in the thick ascending limb of Henle's loop.

BACKGROUND: A basolateral Ba(2+)-sensitive KCl cotransporter has previously been proposed as participating in basolateral K+ recycling and transepithelial NaCl reabsorption in the thick ascending limb of Henle's loop (TAL). The aim of the present study was to answer the question as to whether this cotransporter plays a role in transepithelial K+ reabsorption and whether dietary Mg(2+) deficiency, known to regulate the KCl cotransporter in erythrocytes, also regulates KCl transport in the TAL. METHODS: The effects of a low-Mg(2+) diet were investigated on urinary and plasma K+ concentration in control mice and Mg(2+)-deficient mice. Transepithelial Na+, Cl- and K+ net fluxes (J(Na), J(Cl), J(K)), determined in isolated perfused TALs with electron probe analysis or cation-exchange high-performance liquid chromatography (HPLC) and electrophysiological parameters (V(te), R(te)), were measured in both animal groups. Expression of transcripts for the KCl cotransporter and its possible regulation by low-Mg(2+) were studied by RT-PCR in microdissected mouse cortical TAL (CTAL) and medullary TAL (MTAL) segments. RESULTS: In isolated perfused CTALs, basolateral Ba(2+) and amiloride induced a large K+ net secretion towards the tubular lumen, paralleled by a 50% decrease in transepithelial NaCl reabsorption. KCC1 transcripts were found in the mouse CTAL and MTAL. A low-Mg(2+) diet led to diminished urinary K+ excretion, lowered plasma K+ concentration and up-regulation of KCC1 transcripts in the TAL. For low-Mg(2+) diet, this upregulation was associated with increased transepithelial K+ reabsorption in the in vitro-perfused CTAL. CONCLUSIONS: Our study provides evidence that the KCl cotransporter, which is functionally expressed in the TAL, plays an important role in transepithelial K+ reabsorption. Direct inhibition of this transporter by Ba(2+) and its indirect inhibition by amiloride lead to a strong transepithelial K+ secretion and diminished NaCl reabsorption in the TAL. Up-regulation of KCC1 mRNA by dietary Mg(2+) restriction is associated with an increased K+ reabsorption in the in vitro perfused CTAL.

Chronic exposure to vasopressin upregulates ENaC and sodium transport in the rat renal collecting duct and lung.

Vasopressin is known to acutely stimulate sodium transport in the renal collecting duct. We investigated the long-term regulation by vasopressin of the epithelial sodium channel (ENaC) in the rat kidney. Five-day infusion of dDAVP (a V(2) receptor agonist) to Brattleboro rats lacking vasopressin induced a marked increase in beta- and gamma-subunit ENaC mRNA levels in the renal cortex (beta, 85%; gamma, 100%), with no change in alpha-ENaC mRNA. Expression of beta- and gamma-ENaC mRNAs was also enhanced in lung (beta, 49%; gamma, 33%) but not in distal colon (an organ devoid of V(2) receptors). Similar results were obtained in Sprague Dawley rats after either partial water restriction or dDAVP infusion for 5 days. Transepithelial voltage and transepithelial sodium and water net fluxes were measured in isolated perfused cortical collecting ducts of Brattleboro rats. Acute addition of 2x10(-10) mol/L dDAVP to the bath increased sodium and water fluxes in the same proportion, and to a far greater extent in dDAVP-infused than in control Brattleboro rats (change in Na(+) net flux, 337+/-30 versus 49+/-11 pmol. min(-1). mm(-1), respectively; P<0.001). These effects were abolished by amiloride. Extrarenal water losses, partly originating from the lung, were reduced by high plasma vasopressin level. This study shows that vasopressin increases sodium transport in the renal collecting duct and probably in the lung, through a differential transcriptional regulation of ENaC subunits. This effect is followed by isoosmotic water reabsorption and likely contributes to the process of water conservation. It could lead to less efficient sodium excretion, however, and thus participate in some forms of salt-sensitive hypertension.

Cellular adaptation of the mouse cortical thick ascending limb of Henle's loop (CTAL) to dietary magnesium restriction: enhanced transepithelial Mg2+ and Ca2+ transport.

Mice aged 4 or 8 weeks were fed with a low-Mg2+ diet for 1, 2, 3 or 4 days. After 1 day of diet, the urinary excretion of Mg2+ and Ca2+ was strongly reduced in both animal groups (4 and 8 weeks), accompanied by a significant fall in plasma Mg2+ concentration and an increase in urinary volume. This profile persisted after 2, 3 or 4 days of dietary Mg2+ restriction. After 1 day of diet, transepithelial ion net fluxes of Na+, Cl-, Ca2+ and Mg2+ (JNa' JCI, JCl, JMg) measured in vitro from isolated perfused cortical thick ascending limbs (CTALs) of these animals remained unchanged. After 2 days of diet, measurements of J(Ca) and J(Mg) in isolated perfused CTALs showed that transepithelial Mg2+ and Ca2+ reabsorption were enhanced in CTALs from Mg(2+)-depleted, 8-week-old animals, whereas transepithelial Mg2+ and Ca2+ transport were not altered in 4-week-old mice. JNa and JCl and the transepithelial potential (PDte) were not modified in CTALs from either animal group. Our results suggest that a low-Mg2+ diet leads to urinary retention of Mg2+ and Ca2+ which is most likely due to increased Mg2+ and Ca2+ transport in the CTAL. Furthermore, in response to dietary Mg2+ restriction, the reabsorption of divalent cations in the CTAL of adult, but not of young, mice undergoes cellular adaptation.