Characterization of 2',7'-dichlorofluorescin fluorescence in dissociated mammalian brain neurons: estimation on intracellular content of hydrogen peroxide.
The fluorescence of 2',7'-dichlorofluorescin (DCF) was measured in acutely dissociated rat cerebellar neurons as a mean of estimating the formation of reactive oxygen species (ROS). N,N-Diethyldithiocarbamate, an inhibitor for superoxide dismutase, reduced the intensity of DCF fluorescence in a dose-dependent fashion at concentrations of 30 nM to up to 10 microM. N-Ethylmaleimide, an inhibitor for glutathione peroxidase, augmented the DCF fluorescence in a dose-dependent manner at concentration of 10 microM to 1 mM while 3-amino-1,2,4-triazole, an inhibitor for catalase, did not change the fluorescence intensity even at concentrations as high as 1 mM. Hydrogen peroxide, applied externally at concentrations between 3 microM and 3 mM, augmented the fluorescence in a dose-dependent fashion. These results suggest the possibility that the DCF fluorescence may be useful in estimating the intracellular content of hydrogen peroxide of mammalian brain neurons.
Oyama
Y
Y
Laboratory of Cell Signaling (Pharmacological Science), Faculty of Integrated Arts and Sciences, University of Tokushima, Japan.
Hayashi
A
A
Ueha
T
T
Maekawa
K
K
D000818:Animals; D002531:Cerebellum / Q000737:chemistry*; D005452:Fluoresceins / Q000032:analysis*; D005453:Fluorescence; D006861:Hydrogen Peroxide / Q000032:analysis*; D009474:Neurons / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species
D005452:Fluoresceins; D017382:Reactive Oxygen Species; C029569:diacetyldichlorofluorescein; C037631:2',7'-dichlorofluorescein; D006861:Hydrogen Peroxide