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Journal archive > 2007 > N 6 November-December

INVESTIGATION OF Ca2+-INDUCED CHANGES OF MEMBRANE POTENTIAL
OF SMOOTH MUSCLE MITOCHONDRIA USING FLOW CYTOMETRIC ANALYSIS

L. G. Babich, S. G. Shlykov, N. V. Naumova, S. O. Kosterin

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
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Using flow cytometric analysis and potential-sensitive fluorescent dye TMRM Ca2+-induced changes of membrane potential of isolated smooth muscle mitochondria were studied. It was shown, that Ca2+ (100 µM) addition to the incubation medium induced mitochondrial membrane depolarization that probably could be explained by Са2++-exchanger activation which functioning lead to membrane potential dissipation. In the case of ruthenium red (10 µM) preliminary presence in incubation medium, Ca2+ (100 µM) addition did not lead to membrane potential dissipation. Hence, membrane potential dissipation was caused by an increase of matrix Ca2+ concentration. In the presence of Mg2+ (3 mM) and ATP (3 mM), Ca2+ addition did not cause depolarization. It was supposed that in this case ATP synthase acted in the opposite direction as Н+-pump and prevented from mitochondrial membrane potential dissipation. Thus, the flow cytometry method allows to register membrane potential of isolated smooth muscle mitochondria and also to test the effectors, capable to modulate this parameter.

Key words: flow cytometry, isolated mitochondria, membrane potential, Ca2+, smooth muscles.

The original article in Ukrainian is available for download in PDF format.

© The Ukrainian Biochemical Journal