Adenosine inhibits the transfected Na+-H+ exchanger NHE3 in Xenopus laevis renal epithelial cells (A6/C1)

Francesca Di Sole, Valeria Casavola, Luca Mastroberardino, François Verrey, Orson W. Moe, Gerhard Burckhardt, Heini Murer, Corinna Helmle-Kolb

Research output: Contribution to journalArticle

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Abstract

1. Adenosine influences the vectorial transport of Na+ and HCO3- across kidney epithelial cells. However, its action on effector proteins, such as the Na+-H+ exchanger NHE3, an epithelial brush border isoform of the Na+-H+ exchanger (NHE) gene family, is not yet defined. 2. The present study was conducted in Xenopus laevis distal nephron A6 epithelia which express both an apical adenosine receptor of the A1 type (coupled to protein kinase C (PKC)) and a basolateral receptor of the A2 type (coupled to protein kinase A (PKA)). The untransfected A6 cell line expresses a single NHE type (XNHE) which is restricted to the basolateral membrane and which is activated by PKA. 3. A6 cell lines were generated which express exogenous rat NHE3. Measurements of side-specific pH(i) recovery from acid loads in the presence of HOE694 (an inhibitor with differential potency towards individual NHE isoforms) detected an apical resistant Na+-H+ exchange only in transfected cell lines. The sensitivity of the basolateral NHE to HOE694 was unchanged, suggesting that exogenous NHE3 was restricted to the apical membrane. 4. Stimulation of the apical A1 receptor with N6-cyclopentyladenosine (CPA) inhibited both apical NHE3 and basolateral XNHE. These effects were mimicked by the addition of the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) and partially prevented by the PKC inhibitor calphostin C which also blocked the effect of PMA. 5. Stimulation of the basolateral A2 receptor with CPA inhibited apical NHE3 and stimulated basolateral XNHE. These effects were mimicked by 8-bromo-cAMP and partially prevented by the PKA inhibitor H89 which entirely blocked the effect of 8-bromo-cAMP. 6. In conclusion, CPA inhibits rat NHE3 expressed apically in A6 epithelia via both the apical PKC-coupled A1 and the basolateral PKA-coupled A2 adenosine receptors.

Original languageEnglish (US)
Pages (from-to)829-842
Number of pages14
JournalJournal of Physiology
Volume515
Issue number3
StatePublished - Mar 15 1999

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Sodium-Hydrogen Antiporter
Xenopus laevis
Adenosine
Cyclic AMP-Dependent Protein Kinases
Epithelial Cells
Protein Kinase C
Kidney
8-Bromo Cyclic Adenosine Monophosphate
Protein Kinase Inhibitors
varespladib methyl
Cell Line
Protein Isoforms
Acetates
Epithelium
Adenosine A2 Receptors
Adenosine A1 Receptors
Membranes
Protein C Inhibitor
Nephrons
Microvilli

ASJC Scopus subject areas

  • Physiology

Cite this

Di Sole, F., Casavola, V., Mastroberardino, L., Verrey, F., Moe, O. W., Burckhardt, G., ... Helmle-Kolb, C. (1999). Adenosine inhibits the transfected Na+-H+ exchanger NHE3 in Xenopus laevis renal epithelial cells (A6/C1). Journal of Physiology, 515(3), 829-842.

Adenosine inhibits the transfected Na+-H+ exchanger NHE3 in Xenopus laevis renal epithelial cells (A6/C1). / Di Sole, Francesca; Casavola, Valeria; Mastroberardino, Luca; Verrey, François; Moe, Orson W.; Burckhardt, Gerhard; Murer, Heini; Helmle-Kolb, Corinna.

In: Journal of Physiology, Vol. 515, No. 3, 15.03.1999, p. 829-842.

Research output: Contribution to journalArticle

Di Sole, F, Casavola, V, Mastroberardino, L, Verrey, F, Moe, OW, Burckhardt, G, Murer, H & Helmle-Kolb, C 1999, 'Adenosine inhibits the transfected Na+-H+ exchanger NHE3 in Xenopus laevis renal epithelial cells (A6/C1)', Journal of Physiology, vol. 515, no. 3, pp. 829-842.
Di Sole F, Casavola V, Mastroberardino L, Verrey F, Moe OW, Burckhardt G et al. Adenosine inhibits the transfected Na+-H+ exchanger NHE3 in Xenopus laevis renal epithelial cells (A6/C1). Journal of Physiology. 1999 Mar 15;515(3):829-842.
Di Sole, Francesca ; Casavola, Valeria ; Mastroberardino, Luca ; Verrey, François ; Moe, Orson W. ; Burckhardt, Gerhard ; Murer, Heini ; Helmle-Kolb, Corinna. / Adenosine inhibits the transfected Na+-H+ exchanger NHE3 in Xenopus laevis renal epithelial cells (A6/C1). In: Journal of Physiology. 1999 ; Vol. 515, No. 3. pp. 829-842.
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AU - Di Sole, Francesca

AU - Casavola, Valeria

AU - Mastroberardino, Luca

AU - Verrey, François

AU - Moe, Orson W.

AU - Burckhardt, Gerhard

AU - Murer, Heini

AU - Helmle-Kolb, Corinna

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N2 - 1. Adenosine influences the vectorial transport of Na+ and HCO3- across kidney epithelial cells. However, its action on effector proteins, such as the Na+-H+ exchanger NHE3, an epithelial brush border isoform of the Na+-H+ exchanger (NHE) gene family, is not yet defined. 2. The present study was conducted in Xenopus laevis distal nephron A6 epithelia which express both an apical adenosine receptor of the A1 type (coupled to protein kinase C (PKC)) and a basolateral receptor of the A2 type (coupled to protein kinase A (PKA)). The untransfected A6 cell line expresses a single NHE type (XNHE) which is restricted to the basolateral membrane and which is activated by PKA. 3. A6 cell lines were generated which express exogenous rat NHE3. Measurements of side-specific pH(i) recovery from acid loads in the presence of HOE694 (an inhibitor with differential potency towards individual NHE isoforms) detected an apical resistant Na+-H+ exchange only in transfected cell lines. The sensitivity of the basolateral NHE to HOE694 was unchanged, suggesting that exogenous NHE3 was restricted to the apical membrane. 4. Stimulation of the apical A1 receptor with N6-cyclopentyladenosine (CPA) inhibited both apical NHE3 and basolateral XNHE. These effects were mimicked by the addition of the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) and partially prevented by the PKC inhibitor calphostin C which also blocked the effect of PMA. 5. Stimulation of the basolateral A2 receptor with CPA inhibited apical NHE3 and stimulated basolateral XNHE. These effects were mimicked by 8-bromo-cAMP and partially prevented by the PKA inhibitor H89 which entirely blocked the effect of 8-bromo-cAMP. 6. In conclusion, CPA inhibits rat NHE3 expressed apically in A6 epithelia via both the apical PKC-coupled A1 and the basolateral PKA-coupled A2 adenosine receptors.

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