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Generation of hydrogen peroxide in the developing rat heart: the role of elastin metabolism

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Abstract

Reports describing production of reactive oxygen species in neonatal heart are missing. As lysyl oxidase is potentially important source of H2O2, we studied its role during ontogenic development of rat heart. H2O2 was detected in thin sections of developing rat heart by fluorescence microscopy with the use of fluorescence probe 2′-7′-dichlorofluorescin. The experimental design comprised foetuses 21 days after conception, and then the animals sampled on the 1st, 4th, 7th, 10th, 15th, 30th and 60th day after birth. We also used 7-month-old animals as an example of ageing effects. Since the day 4 on, H2O2 was produced only extracellularly up to the day 15, between days 30 and 60 intracellular production was detected as well, and in 7-month-old animals only extracellular production was observed. The specific inhibitors of lysyl oxidase almost completely quenched the H2O2-dependent fluorescence. Starting from day 7, blue autofluorescence specific to oxidized proteins developed in the vessel wall. Intracellular blue autofluorescence specific to autoxidation products developed after day 30. Chloroform extraction diminished the intracellular blue fluorescence, leaving the extracellular fluorescence intact. This confirmed the protein nature of the fluorophores. Lysyl oxidase is significant source of H2O2 in the heart vessel wall during development and H2O2 oxidatively modifies elastin producing protein blue autofluorescence.

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Acknowledgments

The study was supported by Grant Agency of Czech Republic, grant no. P303/11/0298.

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Authors and Affiliations

  1. Department of Medical Chemistry and Biochemistry, Centre of Cardiovascular Research, 2nd Medical School, Charles University, Plzeňská 221, 150 00, Prague 5, Czech Republic

    Jiří Wilhelm & Richard Vytášek

  2. Centre of Cardiovascular Research, Institute of Physiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague 4, Czech Republic

    Ivana Ošt’ádalová

  3. Department of Histology and Embryology, 2nd Medical School, Charles University, Plzeňská 221, 150 00, Prague 5, Czech Republic

    Luděk Vajner

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  1. Jiří Wilhelm
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  2. Ivana Ošt’ádalová
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  3. Richard Vytášek
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  4. Luděk Vajner
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Correspondence to Jiří Wilhelm.

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Wilhelm, J., Ošt’ádalová, I., Vytášek, R. et al. Generation of hydrogen peroxide in the developing rat heart: the role of elastin metabolism. Mol Cell Biochem 358, 215–220 (2011). https://doi.org/10.1007/s11010-011-0937-8

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  • DOI: https://doi.org/10.1007/s11010-011-0937-8

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