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Variations in elemental compositions of rat hippocampal formation between acute and latent phases of pilocarpine-induced epilepsy: an X-ray fluorescence microscopy study

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Abstract

There is growing experimental evidence that tracing the elements involved in brain hyperexcitability, excitotoxicity, and/or subsequent neurodegeneration could be a valuable source of data on the molecular mechanisms triggering or promoting further development of epilepsy. The most frequently used experimental model of the temporal lobe epilepsy observed in clinical practice is the one based on pilocarpine-induced seizures. In the frame of this study, the elemental anomalies occurring for the rat hippocampal tissue in acute and silent periods after injection of pilocarpine in rats were compared. X-ray fluorescence microscopy was applied for the topographic and quantitative elemental analysis. The differences in the levels of elements such as P, S, K, Ca, Fe, Cu, and Zn between the rats 3 days (SE72) and 6 h (SE6) after pilocarpine injection as well as naive controls were examined. Comparison of SE72 and control groups showed, for specific areas of the hippocampal formation, lower levels of P, K, Cu, and Zn, and an increase in Ca accumulation. These results as well as further analysis of the differences between the SE72 and SE6 groups confirmed that seizure-induced excitotoxicity as well as mossy fiber sprouting are the mechanisms involved in the neurodegenerative processes which may finally lead to spontaneous seizures in the chronic period of the pilocarpine model. Moreover, in the light of the results obtained, Cu seems to play a very important role in the pathogenesis of epilepsy in this animal model. For all areas analyzed, the levels of this element recorded in the latent period were not only lower than those for controls but were even lower than the levels found in the acute period. The decreased hippocampal accumulation of Cu in the phase of behavior and EEG stabilization, a possible inhibitory effect of this element on excitatory amino acid receptors, and enhanced seizure susceptibility in Menkes disease (an inherited Cu transport disorder leading to Cu deficiency in the brain) suggest a neuroprotective role rather than neurodegenerative and proconvulsive roles of Cu in pilocarpine-induced epilepsy.

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Acknowledgments

This work was partially supported by the Polish Ministry of Science and Higher Education and its grant for scientific research IUVENTUS PLUS no. JP2010005370. The research leading to these results has received funding from the European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 226716 and was realized in the frame of experimental grants DESY-D-II-20080009 EC and I-20110056 EC. The authors wish to express their appreciation to Henryk Figiel for valuable discussions and comments in the preparation of the manuscript.

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Authors and Affiliations

  1. Department of Medical Physics and Biophysics, Faculty of Physics and Applied Computer Science, AGH University of Science and Technology, Kraków, Poland

    J. Chwiej & J. Dulinska

  2. Department of Neuroanatomy, Institute of Zoology, Jagiellonian University, Kraków, Poland

    K. Janeczko & Z. Setkowicz

  3. Deutsches Elektronen-Synchrotron (DESY), Hamburg, Germany

    K. Appel

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  1. J. Chwiej
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  2. J. Dulinska
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  3. K. Janeczko
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  4. K. Appel
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  5. Z. Setkowicz
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Correspondence to J. Chwiej.

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Chwiej, J., Dulinska, J., Janeczko, K. et al. Variations in elemental compositions of rat hippocampal formation between acute and latent phases of pilocarpine-induced epilepsy: an X-ray fluorescence microscopy study. J Biol Inorg Chem 17, 731–739 (2012). https://doi.org/10.1007/s00775-012-0892-1

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