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Different effects of CsA and FK506 on aquaporin-2 abundance in rat primary cultured collecting duct cells

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Abstract

Calcineurin (Cn) inhibitors (CnI) such as cyclosporine A (CsA) and FK506 are nephrotoxic immunosuppressant drugs, which decrease tubular function. Here, we examined the direct effect of CnI on aquaporin-2 (AQP2) expression in rat primary cultured inner medullary collecting duct cells. CsA (0.5–5 μM) but not FK 506 (0.01–1 μM) decreased expression of AQP2 protein and messenger RNA (mRNA) in a concentration and time dependent manner, without affecting mRNA stability. This effect was observed despite similar inhibition of Cn activity by both CnI, thereby suggesting that the CsA-dependent decrease in AQP2 expression was Cn independent. Another inhibitor of cyclophilin A, the primary intracellular target of CsA, had no effect on AQP2 expression. In order to investigate the mechanism of decreased AQP2 transcription, we studied activation status of two suggested transcriptional regulators of AQP2, cAMP-responsive element binding protein (CREB), and tonicity enhancer binding protein (TonEBP). Localization of TonEBP, as well as TonEBP-mediated gene transcription, was not affected by CsA. Phosphorylation of CREB at an activating phosphorylation site (S133) was decreased by CsA, but not by FK506. However, both CnI did not affect cellular cAMP levels. We show that CsA decreases transcription of AQP2, a process that is in part independent of Cn or cyclophilin A and suggests dependence on decreased activity of CREB.

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Acknowledgements

This study was supported by a grant of “Innovative Medical Research,” University of Münster Medical Faculty (ED210709) to B.E., the Else-Kröner Fresenius Stiftung to B.E. and E.S, and a grant “Förderprojekte für Studierende (SAFIR) of the University of Münster” to M.M.R. The technical help of Rita Schröter, Julia Humberg, Bernadette Gelschefarth, and Kathrin Beul is deeply acknowledged.

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The authors have no financial conflict of interest.

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  1. Medizinische Klinik und Poliklinik D, Experimentelle Nephrologie, Universität Münster, Domagkstraße 3a, 48149, Münster, Germany

    Markus M. Rinschen, Jens Klokkers, Hermann Pavenstädt, Ute Neugebauer, Eberhard Schlatter & Bayram Edemir

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  1. Markus M. Rinschen
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Correspondence to Bayram Edemir.

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Rinschen, M.M., Klokkers, J., Pavenstädt, H. et al. Different effects of CsA and FK506 on aquaporin-2 abundance in rat primary cultured collecting duct cells. Pflugers Arch - Eur J Physiol 462, 611–622 (2011). https://doi.org/10.1007/s00424-011-0994-6

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