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Vol. 14, Issue 7, 2677-2688, July 2003
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*Division de Néphrologie, Fondation pour
Recherches Médicales, CH-1211 Genève 4, Switzerland;
Service de Néphrologie
Pédiatrique, Hôpital A. Trousseau, F-75571 Paris cedex 12,
France; Laboratoire de Physiologie et
Génomique des Cellules Rénales, FRE 2468, Institut des
Cordeliers, IFR 58, 75270 Paris cedex 6, France; and
INSERM U478, Faculté de Médecine
Xavier Bichat, BP416, F-75870 Paris Cedex 18, France
Submitted November 11, 2002;
Revised February 13, 2003;
Accepted March 13, 2003
Monitoring Editor: Guido Guidotti
In the mammalian kidney the fine control of Na+ reabsorption takes place in collecting duct principal cells where basolateral Na,K-ATPase provides the driving force for vectorial Na+ transport. In the cortical collecting duct (CCD), a rise in intracellular Na+ concentration ([Na+]i) was shown to increase Na,K-ATPase activity and the number of ouabain binding sites, but the mechanism responsible for this event has not yet been elucidated. A rise in [Na+]i caused by incubation with the Na+ ionophore nystatin, increased Na,K-ATPase activity and cell surface expression to the same extent in isolated rat CCD. In cultured mouse mpkCCDcl4 collecting duct cells, increasing [Na+]i either by cell membrane permeabilization with amphotericin B or nystatin, or by incubating cells in a K+-free medium, also increased Na,K-ATPase cell surface expression. The [Na+]i-dependent increase in Na,K-ATPase cell-surface expression was prevented by PKA inhibitors H89 and PKI. Moreover, the effects of [Na+]i and cAMP were not additive. However, [Na+]i-dependent activation of PKA was not associated with an increase in cellular cAMP but was prevented by inhibiting the proteasome. These findings suggest that Na,K-ATPase may be recruited to the cell membrane following an increase in [Na+]i through cAMP-independent PKA activation that is itself dependent on proteasomal activity.
Abbreviations used: PKA, protein kinase A; CCD, cortical collecting duct; AEBSF, 4-(2-aminoethyl)benzenesulfonyl fluoride.
¶ Corresponding author. E-mail address: Eric.Feraille{at}medecine.unige.ch.
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