Lucigenin (Bis-N-methylacridinium) as a Mediator of Superoxide Anion Production

Liochev, Stefan I.; Fridovich, Irwin

doi:10.1006/abbi.1997.9766

Cited by 207 publications

(115 citation statements)

References 0 publications

Supporting

Mentioning

110

Contrasting

Unclassified

Order By: Relevance

“…The reasons for the difference between our results and those reported by Rembish and Trush (40) are unclear, although many factors may have been important including the instrumentation used to make determinations. There are also suggestions that lucigenin may generate ⅐ O 2 Ϫ through redox cycling reactions, making it unsuitable as an indicator (42,43); however, we observed only minor changes in background when lucigenin was mixed with NADH or NADH and heat-killed cells. This suggests that under the conditions used the interference from lucigenin-generated oxidants was probably minimal.…”

Section: Ratio Of Electron Entry To Exit-contrasting

confidence: 50%

Development and Age-associated Differences in Electron Transport Potential and Consequences for Oxidant Generation

Allen

Keogh²,

Tresini

et al. 1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

We determined the activities of NADH dehydrogenase (ND), succinate dehydrogenase, and cytochrome c oxidase (COX) in 29 skin fibroblast lines established from donors ranging in age from 12 gestational weeks to 94 years. The results of this study demonstrate that all three of the enzyme activities examined are greater in adult-derived fibroblasts than in the fetal cell lines. The ratio of enzyme activities that control electron entry into and exit from the electron transport chain varied directly with lucigenin-detected chemiluminescence (an indicator of ⅐ O 2 ؊ generation) and inversely with H 2 O 2 generation. These results indicate a clear difference in the predominant oxidant species generated during fetal and adult stages of life. We also examined the mRNA abundances of different components of the electron transport chain complexes. We observed higher abundances of mitochondrial encoded mRNAs (COX 1 and ND 4) in cell lines established from adults than in fetal cells. No differences in the mRNA abundances of the nuclear encoded sequences (COX 4 and ND 51) were observed in fetal and postnatal-derived lines. Succinate dehydrogenase mRNA abundance was greater in cell lines established from postnatal donors than in fetal cell lines. No significant differences between cell lines established from young and old adults were detected in any of the parameters examined.Mitochondria are the primary site of aerobic energy production and are thus the major site of generation of reactive oxygen species (ROS), 1 such as oxygen-centered free radicals and peroxides, which are by-products of metabolism (1-4). Estimates of the rate of ROS generation can differ greatly; however, it has been demonstrated repeatedly that the relative rate of oxidant generation increases with age and that this increase correlates strongly with age-associated changes in the cellular redox state (5-8). For example, the rates of superoxide ( ⅐ O 2 Ϫ ) (8 -14) and H 2 O 2 generation (7, 10, 14 -17) increase in the cells of aging organisms, whereas the glutathione concentration declines progressively with advancing age (5,6,18,19). Differences in ROS metabolism also exist between cells obtained from early developmental stages and from postnatal tissues (for review, see Refs. 20 and 21). The activities of enzymic antioxidant defenses tend to be lower in fetal cells than in cells derived from postnatal tissues (22-24); however, the GSH concentration is frequently greater in fetal cells than in cells from adults (20,24). Whether these observations reflect differences in the steadystate level of ROS in fetal and postnatal cells is not presently known (20, 21).The rate of mitochondrial ROS generation in cells is largely dependent on the amounts of autoxidizable respiratory carriers and the redox state of electron carriers (1,3,25,26). All electron carriers located prior to one of low abundance will exhibit a greater propensity to be chemically reduced because of the slowing of electron flux (electron stacking; 1, 16, 25, 27). Thus, either age or development-a...

show abstract

Section: Ratio Of Electron Entry To Exit-contrasting

confidence: 50%

Development and Age-associated Differences in Electron Transport Potential and Consequences for Oxidant Generation

Allen

Keogh²,

Tresini

et al. 1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…For instance, DCFH has been shown to react with agents other than H 2 O 2 making it more of a measure of total redox status rather than cellular H 2 O 2 [54,55]. Moreover, both lucigenin and DCFH have been shown to yield a fluorescent signal in the absence of ROS, giving rise to the potential for high background and false positive readings [56][57][58]. In addition to these limitations, one must also assume that cellular uptake of DCFH-DA is equivalent under different conditions if a direct comparison is to be made.…”

Section: Measurement Of Cellular Rosmentioning

confidence: 99%

Reactive oxygen species and cellular oxygen sensing

Cash

Pan

Simon

2007

Free Radical Biology and Medicine

158

View full text Add to dashboard Cite

Many organisms activate adaptive transcriptional programs to help them cope with decreased oxygen levels, or hypoxia, in their environment. These responses are triggered by various oxygen sensing systems in bacteria, yeast and metazoans. In metazoans, the hypoxia inducible factors (HIFs) mediate the adaptive transcriptional response to hypoxia by upregulating genes involved in maintaining bioenergetic homeostasis. The HIFs in turn are regulated by HIF-specific prolyl hydroxlase activity, which is sensitive to cellular oxygen levels and other factors such as tricarboxylic acid cycle metabolites and reactive oxygen species (ROS). Establishing a role for ROS in cellular oxygen sensing has been challenging since ROS are intrinsically unstable and difficult to measure. However, recent advances in fluorescence energy transfer resonance (FRET)-based methods for measuring ROS are alleviating some of the previous difficulties associated with dyes and luminescent chemicals. In addition, new genetic models have demonstrated that functional mitochondrial electron transport and associated ROS production during hypoxia are required for HIF stabilization in mammalian cells. Current efforts are directed at how ROS mediate prolyl hydroxylase activity and hypoxic HIF stabilization. Progress in understanding this process has been enhanced by the development of the FRET-based ROS probe, an vivo prolyl hydroxylase reporter and various genetic models harboring mutations in components of the mitochondrial electron transport chain.

show abstract

“…Liochev and colleagues 26 have reported that high concentrations of lucigenin may produce redox cycling leading to artificial increases in O 2 Ϫ . The concentration of lucigenin used for our initial studies (250 mmol/L) was relatively high; however, we wanted to be able to compare our results with these previously obtained in young animals.…”

Section: O 2 ؊ Measurementmentioning

confidence: 99%

Superoxide Excess in Hypertension and Aging

et al. 2001

View full text Add to dashboard Cite

Abstract-There is evidence in humans that hypertension and aging similarly impair endothelial function, although the mechanism remains unclear. Superoxide anion (O 2 Ϫ ) is a major determinant of nitric oxide (NO) bioavailability and thus endothelial function. We sought to determine the relationship between endothelial function, O 2 Ϫ , and age in normotensive Wistar-Kyoto (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP). Aortic rings were removed from female WKY and SHRSP at 3 to 4 months (young) and 9 to 12 months (old). O 2 Ϫ generation by aortic rings was measured before and after removal of the endothelium or incubation with N G nitro-L-arginine methyl ester, diphenyleneiodonium, or apocynin. Levels of p22phox were studied with immunohistochemistry and used as a marker of NAD (P) Key Words: endothelium Ⅲ nitric oxide Ⅲ hypertension, experimental Ⅲ aging T here is evidence that in animal models and in humans, impaired endothelial function and a decrease in nitric oxide (NO) bioavailability may occur in hypercholesterolemia, 1,2 , diabetes, 3 and hypertension 4 -6 despite normal or increased NO production by the endothelium. 6 A decrease in NO bioavailability may also occur with aging. [7][8][9][10] In a number of animal models of disease, including hypertension 11,12 and hypercholesterolemia, 13 an increase in superoxide (O 2 Ϫ ) occurs concurrent to the decrease in NO bioavailability. O 2 Ϫ rapidly reacts with NO, forming peroxynitrite and decreasing NO bioavailability. 14 Thus, it has been proposed that elevations in O 2 Ϫ levels contribute to the impaired endothelial function associated with atherosclerotic disease. 13,15 Taddei et al 9 proposed that the endothelial dysfunction that occurs in hypertension represents an accelerated form of the dysfunction that occurs with aging. However, the effects of aging on O 2 Ϫ production are less well defined. Huraux and colleagues 16 observed a negative correlation between O 2 Ϫ levels and age in human internal mammary arteries. In contrast, Berry et al 17 found basal O 2 Ϫ production in human internal mammary arteries to be weakly but positively associated with age. Potential vascular sources of O 2 Ϫ are endothelial NO synthase (eNOS), 18 xanthine oxidase, 19 and NAD(P)H oxidase. 20,21 eNOS 18 and NAD(P)H oxidase 22,23 have been proposed to be involved in O 2 Ϫ production in different models of hypertension, whereas xanthine oxidase may be involved in O 2 Ϫ production in hypercholesterolemia. 13 eNOS can be inhibited by arginine analogues such as N G nitro-L-arginine methyl ester (L-NAME). NAD(P)H oxidase is composed of at least 5 subunits, and apocynin can inhibit enzymatic activity by preventing association of the subunits. Diphenyleneiodonium (DPI) is a less specific inhibitor of flavincontaining oxidases, including NAD(P)H oxidase.In this study, the hypothesis that both hypertension and aging result in increased levels of O 2 Ϫ and decreased NO bioavailability in blood vessels from Wistar-Kyoto rats (WKY) and stroke-prone spontaneously hypertens...

show abstract

Lucigenin (Bis-N-methylacridinium) as a Mediator of Superoxide Anion Production

Cited by 207 publications

References 0 publications

Development and Age-associated Differences in Electron Transport Potential and Consequences for Oxidant Generation

Development and Age-associated Differences in Electron Transport Potential and Consequences for Oxidant Generation

Reactive oxygen species and cellular oxygen sensing

Superoxide Excess in Hypertension and Aging

Contact Info

Product

Resources

About