Flow Cytometric Analysis of the H2O2-Induced Increase in Intracellular Ca2+ Concentration of Rat Thymocytes.

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  • Okazaki Eisuke
    Laboratory of Cell Signaling (Pharmacology), Faculty of Integrated Arts and Sciences, The University of Tokushima
  • Chikahisa Lumi
    Laboratory of Cell Signaling (Pharmacology), Faculty of Integrated Arts and Sciences, The University of Tokushima Anticancer and Antimicrobials Research Laboratory, Taiho Pharmaceutical Co., Ltd. Cancer Research Laboratory, Taiho Pharmaceutical Co., Ltd.
  • Kanemaru Kaori
    Laboratory of Cell Signaling (Pharmacology), Faculty of Integrated Arts and Sciences, The University of Tokushima
  • Oyama Yasuo
    Laboratory of Cell Signaling (Pharmacology), Faculty of Integrated Arts and Sciences, The University of Tokushima

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  • low cytometric analysis of the H2O2-induced increase in intracellular Ca2+ concentration of rat thymocytes

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Abstract

The effect of hydrogen peroxide (H2O2) on the intracellular Ca2+ concentration ([Ca2+]i) of rat thymocytes was examined by a flow cytometer and two fluorescent dyes, fluo-3-AM and ethidium bromide, a dye impermeant to intact membranes, to characterize the H2O2-induced increase in [Ca2+]i. H2O2 at concentrations greater than 30 μM dose-dependently increased the [Ca2+]i, of thymocytes which were not stained with ethidium. Removal of external Ca2+ greatly reduced the degree of H2O2-induced increase in [Ca2+]i. However, H2O2 still increased the [Ca2+]i under the external Ca2+-free condition. Diethylmaleate, which is known to produce a chemical depletion of cellular nonprotein thiol, significantly increased the [Ca2+]i. Dithiothreitol, which is used to protect cellular nonprotein thiol, slightly decreased the [Ca2+]i, but greatly reduced the H2O2-induced increase in [Ca2+]i. Therefore, it is considered that H2O2 may increase the [Ca2+]i through a mechanism related to the effects of H2O2 on the cellular nonprotein thiol.

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