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The Ca2+ threshold for the mitochondrial permeability transition and the content of proteins related to Bcl-2 in rat liver and Zajdela hepatoma mitochondria

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Abstract

Zajdela hepatoma mitochondria were able to accumulate two to five times more Ca2+ than rat liver mitochondria before the permeability transition was induced. Pulses of Ca2+ were given in series to determine the Ca2+ threshold by recording changes in [Ca2+] and membrane potential, the permeability transition causing the release of accumulated Ca2+ and collapse of the membrane potential. Hepatoma mitochondria had lower Ca2+ efflux rates, higher net Ca2+ uptake rates and lower phosphorylation rates than liver mitochondria. Since the differences in regard to induction of the permeability transition might be due to higher expression of the Bcl-2 protein in hepatoma cells than in hepatocytes, the transcription of Bcl-2 and the proteins reacting with a Bcl-2 polyclonal antiserum were estimated by Northern and Western blotting, respectively. Hepatoma cells had two Bcl-2 specific mRNA bands of 7 and 2.4 kb, and substantial amounts of the Bcl-2 protein, whereas in liver cells and mitochondria these were not detected. Both cell lines had a reactive band at 19-20 kDa, and hepatocytes a small band at 31-32 kDa. Bcl-2 antibodies stimulated the permeability transition potently in hepatoma mitochondria.

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Authors and Affiliations

  1. Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Russia

    Yuri V. Evtodienko, Vera V. Teplova, Tamara S. Azarashvily, Alexei Kudin & Olga Prusakova

  2. Department of Anatomy, Institute of Biomedicine, University of Helsinki, Finland

    Ismo Virtanen & Nils-Erik L. Saris

Authors
  1. Yuri V. Evtodienko
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  2. Vera V. Teplova
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  3. Tamara S. Azarashvily
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  4. Alexei Kudin
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  5. Olga Prusakova
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  6. Ismo Virtanen
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  7. Nils-Erik L. Saris
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Evtodienko, Y.V., Teplova, V.V., Azarashvily, T.S. et al. The Ca2+ threshold for the mitochondrial permeability transition and the content of proteins related to Bcl-2 in rat liver and Zajdela hepatoma mitochondria. Mol Cell Biochem 194, 251–256 (1999). https://doi.org/10.1023/A:1006913526643

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