Flow Cytometric Analysis of the H_2O_2-Induced Increase in Intracellular Ca^<2+> Concentration of Rat Thymocytes

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Author(s)

    • 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
    • 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

Abstract

The effect of hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>) on the intracellular Ca<SUP>2+</SUP> concentration ([Ca<SUP>2+</SUP>]<SUB>i</SUB>) 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 H<SUB>2</SUB>O<SUB>2</SUB>-induced increase in [Ca<SUP>2+</SUP>]<SUB>i</SUB>. H<SUB>2</SUB>O<SUB>2</SUB> at concentrations greater than 30 μM dose-dependently increased the [Ca<SUP>2+</SUP>]<SUB>i</SUB>, of thymocytes which were not stained with ethidium. Removal of external Ca<SUP>2+</SUP> greatly reduced the degree of H<SUB>2</SUB>O<SUB>2</SUB>-induced increase in [Ca<SUP>2+</SUP>]<SUB>i</SUB>. However, H<SUB>2</SUB>O<SUB>2</SUB> still increased the [Ca<SUP>2+</SUP>]<SUB>i</SUB> under the external Ca<SUP>2+</SUP>-free condition. Diethylmaleate, which is known to produce a chemical depletion of cellular nonprotein thiol, significantly increased the [Ca<SUP>2+</SUP>]<SUB>i</SUB>. Dithiothreitol, which is used to protect cellular nonprotein thiol, slightly decreased the [Ca<SUP>2+</SUP>]<SUB>i</SUB>, but greatly reduced the H<SUB>2</SUB>O<SUB>2</SUB>-induced increase in [Ca<SUP>2+</SUP>]<SUB>i</SUB>. Therefore, it is considered that H<SUB>2</SUB>O<SUB>2</SUB> may increase the [Ca<SUP>2+</SUP>]<SUB>i</SUB> through a mechanism related to the effects of H<SUB>2</SUB>O<SUB>2</SUB> on the cellular nonprotein thiol.

Journal

  • The Japanese Journal of Pharmacology

    The Japanese Journal of Pharmacology 71(4), 273-280, 1996-08-01

    The Japanese Pharmacological Society

References:  28

Cited by:  3

Codes

  • NII Article ID (NAID)
    10008189146
  • NII NACSIS-CAT ID (NCID)
    AA00691188
  • Text Lang
    ENG
  • Article Type
    Journal Article
  • ISSN
    00215198
  • Data Source
    CJP  CJPref  J-STAGE 
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