Pflügers Archiv

, Volume 449, Issue 1, pp 66–75 | Cite as

Stimulation of Xenopus P2Y1 receptor activates CFTR in A6 cells

  • L. Guerra
  • M. Favia
  • T. Fanelli
  • G. Calamita
  • M. Svelto
  • A. Bagorda
  • K. A. Jacobson
  • S. J. Reshkin
  • V. Casavola
Ion Channels, Transporters
First Online:
Received:
Accepted:

Abstract

Nucleotide binding to purinergic P2Y receptors contributes to the regulation of a variety of physiological functions in renal epithelial cells. Here, we investigate the regulatory mechanism of the P2Y1 receptor agonist 2-methylthioadenosine diphosphate (2-MeSADP) on Cl− transport in A6 cells, a commonly used model of the distal section of the Xenopus laevis nephron. Protein and mRNA expression analysis together with functional measurements demonstrated the basolateral location of the Xenopus P2Y1 receptor. 2-MeSADP increased intracellular [Ca2+] and cAMP and Cl− efflux, responses that were all inhibited by the specific P2Y1 receptor antagonist MRS 2179. Cl− efflux was also inhibited by the cystic fibrosis transmembrane conductance regulator (CFTR) blocker glibenclamide. Inhibition of either protein kinase A (PKA) or the binding between A-kinase-anchoring proteins (AKAPs) and the regulatory PKA RII subunit blocked the 2-MeSADP-induced activation of CFTR, suggesting that PKA mediates P2Y1 receptor regulation of CFTR through one or more AKAPs. Further, the truncation of the PDZ1 domain of the scaffolding protein Na+/H+ exchanger regulatory factor-2 (NHERF-2) inhibited 2-MeSADP-dependent stimulation of Cl− efflux, suggesting the involvement of this scaffolding protein. Activation or inhibition of PKC had no effect per se on basal Cl− efflux but potentiated or reduced the 2-MeSADP-dependent stimulation of Cl− efflux, respectively. These data suggest that the X. laevis P2Y1 receptor in A6 cells can increase both cAMP/PKA and Ca2+/PKC intracellular levels and that the PKC pathway is involved in CFTR activation via potentiation of the PKA pathway.

Keywords

Xenopus P2Y1 CFTR NHERF-2 A6 
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Notes

Acknowledgements

This work was supported by grants from: Telethon, Italy, grant E.1125; the Italian Cystic Fibrosis Research Foundation and CEGBA (Centro di Eccellenza di Genomica in Campo Biomedico ed Agrario). We thank Dr. Pann-Ghill Suh of Life Science and School of Environmental Engineering, Pohang University of Science and Tecnology, Pohang 790-784, South Korea, for the kind gift of NHERF-2-ΔPDZ1.

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Copyright information

© Springer-Verlag  2004

Authors and Affiliations

  • L. Guerra
    • 1
  • M. Favia
    • 1
  • T. Fanelli
    • 1
  • G. Calamita
    • 1
  • M. Svelto
    • 1
  • A. Bagorda
    • 1
  • K. A. Jacobson
    • 2
  • S. J. Reshkin
    • 1
  • V. Casavola
    • 1
  1. 1.Department of General and Environmental PhysiologyUniversity of BariBariItaly
  2. 2.Laboratory of Bioorganic Chemistry, NIDDKNational Institutes of HealthBethesdaUSA

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