Localization of damage induced by reactive oxygen species in cultured cells

Free Radical Biology & Medicine
S M VroegopS E Buxser

Abstract

N18-RE-105 neuron-derived hybridoma cells were employed to determine the location and degree of damage induced by each of three reactive oxygen species (ROS) generators: 6-hydroxydopamine (6-OHDA), H2O2, and cumene hydroperoxide. Two readily distinguishable plasma membrane markers were used to assess cell surface damage, namely the active transport of alpha-aminoisobutyric acid (AIB) and the facilitated diffusion of glucose. In addition, staining of mitochondria with a tetrazolium dye, 3[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), was used as an intracellular marker to measure the integrity of the metabolic function of the mitochondria. The dose-response curve of inactivation of transport or of metabolic function varied with the ROS generator used and conformed to one of two patterns of toxicity: either threshold-dependent or single-hit inactivation. We determined that 6-OHDA acts simultaneously on multiple targets and steps in the cells, resulting in a very steep dose-effect curve. Similarly, damage induced by H2O2 to the AIB transporters and to mitochondria is consistent with simultaneous inactivation of multiple steps, but damage to glucose transporters conforms to single-hit inactivation of the transpor...Continue Reading

References

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Citations

May 22, 1995·Chemistry and Physics of Lipids·D E DeckerS E Buxser
Jan 1, 1997·Free Radical Biology & Medicine·G J FiciP F VonVoigtlander
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Nov 10, 2020·The Journal of Veterinary Medical Science·Doo Hyun KimMasayuki Funaba
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Oct 11, 2021·Cell Death and Differentiation·Timothy J HumptonKaren H Vousden

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