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Relationship between 1-chloro2,4-dinitrobenzene-induced cytoskeletal perturbations and cellular glutathione

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Abstract

Exposure of 3T3 cells to micromolar doses of 1-chloro-2,4-dinitrobenzene, a substrate for glutathione-S-transferase, resulted in a rapid depletion of total cellular glutathione accompanied by disassembly of microtubules as visualized by fluorescence microscopy. However, prolonged incubation resulted in cellular recovery from 1-chloro-2,4-dinitrobenzene insult as evidenced by a steady rise in total cellular glutathione accompanied by microtubule reassembly to their normal organization 5 hours after treatment. To evaluate the role of total cellular glutathione in modulating the 1chloro-2,4-dinitrobenzene-induced cytoskeletal perturbation, we used 1-chloro-2,4-dinitrobenzene and/or buthiomine sulfoximine, an effective irreversible inhibitor of glutathione synthesis, to manipulate cellular glutathione levels. Incubation of 3T3 cells with 2.5 μM 1-chloro-2,4-dinitrobenzene and 250 μM buthiomine sulfoximine for 5 hours resulted in a complete depletion of total cellular glutathione accompanied by essentially complete loss of microtubules and marked alterations in the density and distribution pattern of microfilaments. Buthionine sulfoximine enhanced markedly the extent and duration of cellular glutathione depletion and the severity of microtubule disruption of 3T3 cells over the level achieved by 1-chloro-2,4-dinitrobenzene treatment alone. Furthermore, buthiomine sulfoximine also prevented the restoration of cellular glutathione content and microtubule reassembly that normally were evident 5 hours after 1-chloro-2,4-dinitrobenzene treatment. Exposure of 3T3 cells to 50 μM 2-cyclohexene-l-one, which depletes free glutathione by conjugation, resulted in a com

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Abbreviations

BSO:

DL-buthiomine-S-R-sulfoximine

CDNB:

1-chloro-2,4-dinitrobenzene

CHX:

2-cyclohexene-l-one

GSH:

glutathione

GST:

glutathione-S-transferase

MAPS:

microtubule-associated proteins

MF:

microfilaments

MT:

microtubules.

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Authors and Affiliations

  1. Department of Microbiology, Boston University School of Medicine, Boston, Massachusetts

    Mun-Fai Leung & Iih-Nan Chou

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  1. Mun-Fai Leung
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  2. Iih-Nan Chou
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Leung, MF., Chou, IN. Relationship between 1-chloro2,4-dinitrobenzene-induced cytoskeletal perturbations and cellular glutathione. Cell Biol Toxicol 5, 51–66 (1989). https://doi.org/10.1007/BF00141064

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  • DOI: https://doi.org/10.1007/BF00141064

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