Mitochondrial transmembrane potential is diminished in phorbol myristate acetate-stimulated peritoneal resident macrophages isolated from wild-type mice, but not in those from gp91-phox-deficient mice

Histochemistry and Cell Biology
Toshihiro KobayashiHarumichi Seguchi

Abstract

Macrophages produce superoxide (O2-) during phagocytosis or upon stimulation with a variety of agents including phorbol myristate acetate (PMA) through the activation of NADPH oxidase, and the formed O2- is converted to other reactive oxygen species (ROS) such as hydrogen peroxide (H2O2). The aim of the present study was to elucidate the effect of the intracellularly produced ROS on mitochondrial transmembrane potential (MTP) in mouse (C57BL/6) peritoneal resident macrophages stimulated with PMA. Using a fluorescent dye, succinimidyl ester of dichlorodihydrofluorescein (H2DCFDA), O2- was visualized in intracellular compartments in a certain subpopulation of macrophages isolated from wild-type mice. Cells deficient in gp91-phox, one of the membrane components of NADPH oxidase, were negative for the fluorescence. When cells were loaded with both H2DCFDA and MitoCapture, a fluorescent dye for mitochondria, mitochondrial fluorescence was diminished in O2- -producing cells, but not in O2- -deficient cells. Flow cytometry also revealed the decrease of mitochondrial fluorescence in wild-type cells, but not in gp91-phox-deficient cells. The loss of mitochondrial fluorescence was prevented by microinjection of catalase into cells. The p...Continue Reading

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Citations

Dec 23, 2004·Histochemistry and Cell Biology·Esther Asan, Detlev Drenckhahn
Nov 12, 2005·Histochemistry and Cell Biology·Douglas J Taatjes, Jürgen Roth
Dec 7, 2006·Histochemistry and Cell Biology·Douglas J TaatjesJürgen Roth
Nov 1, 2007·Histochemistry and Cell Biology·Christian ZuberJürgen Roth
Dec 22, 2014·European Journal of Pharmacology·Csenge AntusBalazs Veres
Nov 3, 2006·The Journal of Nutritional Biochemistry·Denis LaironDominique Jourdheuil-Rahmani
Sep 21, 2007·Journal of Cellular Biochemistry·Hirohiko OkamuraTatsuji Haneji

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