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Presence of organic anion transporters 3 (OAT3) and 4 (OAT4) in human adrenocortical cells

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Abstract

Since the organic anion transporter-1 (OAT1) has been implicated in cortisol release from bovine and rat adrenal zona fasciculata cells, we addressed the question of whether OATs are present in human adrenal cortical cells. In the human adrenal cell line NCI-H295R, 24-h cortisol secretion increased up to 30-fold on exposure to forskolin. Incubation of forskolin-treated cells for 24 h with the OAT substrates probenecid, p-aminohippurate (PAH), glutarate or cimetidine inhibited cortisol release partly. RT-PCR did not reveal expression of human OAT1 and OAT2, but OAT3 and OAT4 mRNAs were detected in both NCI-H295R cells and human adrenal tissue. When human OAT3 (hOAT3) and hOAT4 were expressed in Xenopus laevis oocytes, only hOAT3 showed [3H]cortisol uptake in excess of that of water-injected control oocytes. Cortisol uptake via OAT3 was saturable with an apparent Kt of 2.4 µM. In NCI-H295R cells, [3H]estrone sulphate uptake was saturable, cis-inhibited by OAT substrates and trans-stimulated by preloading with glutarate or cortisol. Likewise, [3H]PAH uptake was cis-inhibited by estrone sulphate and trans-stimulated by preloading the cells with PAH, glutarate or cortisol, indicating functional expression of OATs in the plasma membrane of NCI-H295R cells.

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Acknowledgements

We thank S. Schindelmann for excellent technical assistance, E. Thelen for expert help with the preparation of the illustrations, N. Bakhiya for the injection of oocytes with hOAT3 cRNA and C. Langenberg for organizing the human tissue. This work was supported by Deutsche Forschungsgemeinschaft Grant Bu 571/7-5 and GK335

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Authors and Affiliations

  1. Zentrum für Physiologie und Pathophysiologie, Abt. Vegetative Physiologie und Pathophysiologie, Georg-August-Universität Göttingen, Humboldtallee 23, 37073, Göttingen, Germany

    Andrew Bahn, Gerhard Burckhardt & Yohannes Hagos

  2. Abt. Nephrologie und Rheumatologie, Georg-August-Universität Göttingen, Göttingen, Germany

    Abdul R. Asif, Jürgen Steffgen, R. Willi Grunewald & Gerhard A. Müller

  3. Abt. Klinische und Experimentelle Endokrinologie, Georg-August-Universität Göttingen, Göttingen, Germany

    Maria Metten

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  1. Abdul R. Asif
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  2. Jürgen Steffgen
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  3. Maria Metten
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  4. R. Willi Grunewald
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  5. Gerhard A. Müller
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  6. Andrew Bahn
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  7. Gerhard Burckhardt
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  8. Yohannes Hagos
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Correspondence to Yohannes Hagos.

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Asif, A.R., Steffgen, J., Metten, M. et al. Presence of organic anion transporters 3 (OAT3) and 4 (OAT4) in human adrenocortical cells. Pflugers Arch - Eur J Physiol 450, 88–95 (2005). https://doi.org/10.1007/s00424-004-1373-3

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  • DOI: https://doi.org/10.1007/s00424-004-1373-3

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