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Formation of cysts by principal-like MDCK cells depends on the synergy of cAMP- and ATP-mediated fluid secretion

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Abstract

It has been suggested that more than 70% of the renal cysts in patients with autosomal dominant polycystic kidney disease (ADPKD) arise from the collecting duct and that within this segment cysts originate almost exclusively from principal rather than intercalated cells. The mechanisms for this predisposition of principal cells have so far remained elusive. We, therefore, used Madin–Darby canine kidney (MDCK) subclones resembling principal cells and alpha-intercalated cells in a three-dimensional in vitro model to determine differences in cystogenesis and cyst growth, including the response to cyclic adenosine monophosphate (cAMP) elevation and the dependence on ATP signaling. We found that in vitro cysts developed only from principal-like but not from intercalated-like MDCK cell clones. This specificity could be verified in mixed MDCK cultures enriched for principal- or intercalated-like cells. In vitro cyst growth upon elevation of intracellular cAMP was mainly driven by fluid secretion, rather than increased cell proliferation. The cAMP-dependent fluid secretion was found to depend on extracellular adenosine-5'-triphosphate (ATP) and to act synergistically with purinergic signaling, as the use of the ATP scavenger apyrase, as well as the P2 receptor inhibitor suramin, reduced cAMP-driven fluid secretion, while increasing extracellular ATP potentiated cAMP-mediated cyst growth. In conclusion, we provide in vitro evidence for the ability of principal rather than intercalated cells to form cysts, based on a synergism of cAMP and ATP signaling in enhancing apical fluid secretion.

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Acknowledgments

We thank Hans Oberleithner and Marianne Wilhelmi from the University of Muenster for the kind gift of the C7 and C11 clones. We also thank Margarete Goppelt-Struebe and Carsten Willam for their helpful discussions and technical support and Michael Köttgen for his critical reading of the manuscript. B. Buchholz was a recipient of a stipend from the ELAN-Fonds (Erlanger Leistungsbezogene Anschubfinanzierung und Nachwuchsförderung) and is supported by a junior career development grant of the Interdisciplinary Center of Clinical Research at the University of Erlangen-Nuremberg.

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Authors and Affiliations

  1. Department of Nephrology and Hypertension, Friedrich-Alexander University of Erlangen-Nuremberg, Krankenhausstrasse 12, D-91054, Erlangen, Germany

    Bjoern Buchholz, Barbara Teschemacher, Gunnar Schley & Kai-Uwe Eckardt

  2. Medical Faculty, Institute of Physiology II, University of Muenster, D-48149, Muenster, Germany

    Hermann Schillers

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  1. Bjoern Buchholz
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  2. Barbara Teschemacher
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  3. Gunnar Schley
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  4. Hermann Schillers
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  5. Kai-Uwe Eckardt
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Correspondence to Bjoern Buchholz or Kai-Uwe Eckardt.

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Buchholz, B., Teschemacher, B., Schley, G. et al. Formation of cysts by principal-like MDCK cells depends on the synergy of cAMP- and ATP-mediated fluid secretion. J Mol Med 89, 251–261 (2011). https://doi.org/10.1007/s00109-010-0715-1

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  • DOI: https://doi.org/10.1007/s00109-010-0715-1

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