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Ouabain Regulates CFTR-Mediated Anion Secretion and Na,K-ATPase Transport in ADPKD Cells

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Abstract

Cyst enlargement in autosomal dominant polycystic kidney disease (ADPKD) requires the transepithelial secretion of fluid into the cyst lumen. We previously showed that physiological amounts of ouabain enhance cAMP-dependent fluid secretion and cyst growth of human ADPKD cyst epithelial cells in culture and formation of cyst-like dilations in metanephric kidneys from Pkd1 mutant mice. Here, we investigated the mechanisms by which ouabain promotes cAMP-dependent fluid secretion and cystogenesis. Ouabain (3 nM) enhanced cAMP-induced cyst-like dilations in embryonic kidneys from Pkd1 m1Bei mice, but had no effect on metanephroi from Pkd1 m1Bei mice that lack expression of the cystic fibrosis transmembrane conductance regulator (CFTR). Similarly, ouabain stimulation of cAMP-induced fluid secretion and in vitro cyst growth of ADPKD cells were abrogated by CFTR inhibition, showing that CFTR is required for ouabain effects on ADPKD fluid secretion. Moreover, ouabain directly enhanced the cAMP-dependent Cl− efflux mediated by CFTR in ADPKD monolayers. Ouabain increased the trafficking of CFTR to the plasma membrane and up-regulated the expression of the CFTR activator PDZK1. Finally, ouabain decreased plasma membrane expression and activity of the Na,K-ATPase in ADPKD cells. Altogether, these results show that ouabain enhances net fluid secretion and cyst formation by activating apical anion secretion via CFTR and decreasing basolateral Na+ transport via Na,K-ATPase. These results provide new information on the mechanisms by which ouabain affects ADPKD cells and further highlight the importance of ouabain as a non-genomic stimulator of cystogenesis in ADPKD.

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Acknowledgments

This study was supported by a National Institutes of Health Grant DK081431 to G.B. We thank the Polycystic Kidney Disease Foundation and the PKD Research Biomaterials and Cellular Models Core at the University of Kansas Medical Center and hospitals participating in the Polycystic Kidney Research Retrieval Program for providing human ADPKD kidneys from which primary cells were obtained.

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Authors and Affiliations

  1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS, 66160, USA

    Kyle Jansson, Jessica Venugopal, Gladis Sánchez, Darren P. Wallace & Gustavo Blanco

  2. Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, USA

    Brenda S. Magenheimer & James P. Calvet

  3. Department of Medicine, University of Kansas Medical Center, Kansas City, KS, USA

    Gail A. Reif & Darren P. Wallace

  4. The Kidney Institute, University of Kansas Medical Center, Kansas City, KS, USA

    Kyle Jansson, Jessica Venugopal, Gladis Sánchez, Brenda S. Magenheimer, Gail A. Reif, Darren P. Wallace, James P. Calvet & Gustavo Blanco

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  1. Kyle Jansson
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  2. Jessica Venugopal
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  3. Gladis Sánchez
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Correspondence to Gustavo Blanco.

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Jansson, K., Venugopal, J., Sánchez, G. et al. Ouabain Regulates CFTR-Mediated Anion Secretion and Na,K-ATPase Transport in ADPKD Cells. J Membrane Biol 248, 1145–1157 (2015). https://doi.org/10.1007/s00232-015-9832-7

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