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Transmission electron microscopic and X-ray absorption fine structure spectroscopic investigation of U repartition and speciation after accumulation in renal cells

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Abstract

After environmental contamination, U accumulates in the kidneys and in bones, where it causes visible damage. Recent in vitro data prove that the occurrence of citrate increases U bioavailability without changing its speciation. Two hypotheses can explain the role of citrate: it either modifies the U intracellular metabolization pathway, or it acts on the transport of U through cell membrane. To understand which mechanisms lead to increased bioavailability, we studied the speciation of U after accumulation in NRK-52E kidney cells. U speciation was first identified in various exposure media, containing citrate or not, in which U was supplied as U carbonate. The influence of serum proteins was analyzed in order to detect the formation of macromolecular complexes of U. Transmission electron microscopy (TEM) was employed to follow the evolution of the U species distribution among precipitated and soluble forms. Finally, extended X-ray absorption fine structure spectroscopy (EXAFS) enabled the precipitates observed to be identified as U-phosphate. It also demonstrated that the intracellular soluble form of U is U carbonate. These results suggest that citrate does not change U metabolization but rather plays a role in the intracellular accumulation pathway. U speciation inside cells was directly and clearly identified for the first time. These results elucidate the role of U speciation in terms of its bioavailability and consequent health effects.

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Acknowledgments

This work was partly supported by the Environmental and Nuclear Toxicology program. We thank J. M. Verbavatz for TEM observations, and J. L. Hazemann for helpful discussions during the EXAFS experiments.

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Authors and Affiliations

  1. Laboratoire Pierre Süe CEA-CNRS, CEA/Saclay, 91191, Gif sur Yvette, France

    Marie Carrière, Sarah Milgram, Céline Thiebault, Laure Avoscan & Barbara Gouget

  2. Laboratoire de Géophysique Interne et Tectonophysique, UMR CNRS/Université Joseph Fourier, 1381 rue de la piscine, 38400, Saint-Martin-d’Hères, France

    Olivier Proux

  3. Institut de Physique Nucléaire, 15 rue Georges Clémenceau, 91406, Orsay, France

    Nicole Barre

  4. Laboratoire de Conception des Architectures Moléculaires, CEA/Valrhô, 30207, Bagnols sur Cèze, France

    Christophe Den Auwer

Authors
  1. Marie Carrière
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  2. Olivier Proux
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  3. Sarah Milgram
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  4. Céline Thiebault
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  5. Laure Avoscan
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  6. Nicole Barre
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  7. Christophe Den Auwer
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  8. Barbara Gouget
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Correspondence to Marie Carrière.

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Carrière, M., Proux, O., Milgram, S. et al. Transmission electron microscopic and X-ray absorption fine structure spectroscopic investigation of U repartition and speciation after accumulation in renal cells. J Biol Inorg Chem 13, 655–662 (2008). https://doi.org/10.1007/s00775-008-0350-2

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  • DOI: https://doi.org/10.1007/s00775-008-0350-2

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