Direct measurement of oscillatory generation of superoxide anions by single phagocytes

Isogai, Yasuhiro; Tsuyama, Tomomi; Osada, Hideo; Iizuka, Tetsutarō; Tanaka, Kazuko

doi:10.1016/0014-5793(96)00068-3

Cited by 15 publications

(14 citation statements)

References 15 publications

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“…Another study used a microelectrode to measure the amount of superoxide (O 2 Ϫ ) present within the PMN-like cell line HL-60 (12). It was reported that there was about 15 Ϯ 5 fmol O 2 Ϫ present per cell.…”

Section: Discussionmentioning

confidence: 99%

Salmonella enterica Serovar Typhimurium NiFe Uptake-Type Hydrogenases Are Differentially Expressed In Vivo

Zbell

Maier

2008

Infect Immun

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Salmonella enterica serovar Typhimurium, a common enteric pathogen, possesses three NiFe uptake-type hydrogenases. The results from mouse infection studies suggest that the H 2 oxidation capacity provided by these hydrogenases is important for virulence. Since the three enzymes are similar in structure and function, it may be expected that they are utilized under different locations and times during an infection. A recombination-based method to examine promoter activity in vivo (RIVET) was used to determine hydrogenase gene expression in macrophages, polymorphonuclear leukocyte (PMN)-like cells, and a mouse model of salmonellosis. The hyd and hya promoters showed increased expression in both murine macrophages and human PMN-like cells compared to that in the medium-only controls. Quantitative reverse transcription-PCR results suggested that hyb is also expressed in phagocytes. A nonpolar hya mutant was compromised for survival in macrophages compared to the wild type. This may be due to lower tolerance to acid stress, since the hya mutant was much more acid sensitive than the wild type. In addition, hya mutant cells were internalized by macrophages the same as wild-type cells. Mouse studies (RIVET) indicate that hyd is highly expressed in the liver and spleen early during infection but is expressed poorly in the ileum in infected animals. Late in the infection, the hyd genes were expressed at high levels in the ileum as well as in the liver and spleen. The hya genes were expressed at low levels in all locations tested. These results suggest that the hydrogenases are used to oxidize hydrogen in different stages of an infection.

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Section: Discussionmentioning

confidence: 99%

Salmonella enterica Serovar Typhimurium NiFe Uptake-Type Hydrogenases Are Differentially Expressed In Vivo

Zbell

Maier

2008

Infect Immun

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“…Only a few electrochemical studies examined the oxidative burst problem at the single cell level before, and to the best of our knowledge, only the release of superoxide ion has been reported [48,50]. Release of NO has been investigated electrochemically at the single cell level, but not under oxidative stress conditions [51].…”

Section: Resultsmentioning

confidence: 99%

Nitrogen monoxide and oxidative stress: composition and intensity of cellular oxidative bursts cocktail. A study through artificial electrochemical synapses on single human fibroblasts

Amatore¹,

Arbault²,

Bruce³

et al. 2000

Analusis

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Positioning an ultramicroelectrode at micrometric distances from an isolated living cell ensures that any electroactive material released by the cell is collected and analyzed by the electrode surface. The film of extracellular fluid comprised between the cell and the electrode surfaces defines an artificial synaptic cleft of a few hundred femtoliters volume, in which the release of minute molecular amounts of chemicals produces a sudden and important concentration rise. This guarantees the detection of the released species with an extremely high signalto-noise ratio, as well as a determination of its instant released flux since the collection efficiency is quantitative. In other words, the assembly cell/liquid cleft/ultramicroelectrode behaves as an artificial neuronal synapse. In this review we wish to elaborate on how this artificial synaptic arrangement may be used to monitor an oxidative stress response stimulated in a human fibroblast, and demonstrate that such long-time conjectured oxidative cellular bursts involve a subtle cocktail of femtomoles of hydrogen peroxide, nitrogen monoxide, peroxynitrite and nitrite ions. The analysis performed establishes that this delicate cocktail results from the ultimate combinations of equimolar primary productions of superoxide ion and nitrogen monoxide by two distinct enzymatic systems which are presumably operating in distinct compartments of the cell: NADPH-oxidase type enzymes for the generation of superoxide, and NO-synthases for that of nitrogen monoxide.Article available at http://analusis.edpsciences.org or http://dx.doi.org/10. 1051/analusis:2000280506 Electrodes pick up electrical noise through their capacitance, viz., through their overall conducting surface area, while the analytical information (viz., the Faradaic current) may arise only from the surface area exposed to the cell release. Thus, using an electrode with an active surface matching that of the examined cell decreases the noise while it does not affect at all the quality and intensity of the analytical information. This increases the signal-to-noise ratio and ensures simultaneously a quantitative collection efficiency since the artificial synaptic cleft covers the cell emitting surface. Cells being of micrometric dimensions, it is understood that positioning an ultramicroelectrode at micrometric distances from an isolated living cell ( Fig. 1 a,b) ensures the most adequate signal-to-noise ratio and guarantees that the collection of electroactive chemicals released by a single living cell surface is total [1][2][3][4][5][6] Application of artificial synapses to oxidative stressThe biological problem and its biological and medical importanceAerobic cells are known to produce superoxide ion (O 2 -) and nitrogen monoxide (NO) in response to a stress created by a drastic change in the cell activity provoked by an aggression of its membrane integrity (during a viral or bacterial intrusion for example) or following a sudden and drastic change in its surrounding medium. The released NO is thought to ...

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“…It was based on the oxidation of O 2 -at a carbon microelectrode set at 0.1 V vs SCE and it has allowed to demonstrate periodic fluctuations in O 2 -production by a single phagocytic cell, adsorbed on the opsonized electrode [42]. This study showed a new technique for bringing a single neutrophil cell into a contact with the surface of a microelectrode under the microscope.…”

Section: Non-enzymatic Amperometric Sensorsmentioning

confidence: 99%

“…The carbon fiber is sealed in a glass capillary by heating the glass to melt it around the fibre [40] or by using epoxy resin [8,41,42,44]. The electrode was then polished prior to each run [40] or electrochemically pretreated [8,41].…”

Section: Non-enzymatic Amperometric Sensorsmentioning

confidence: 99%

“…The more efficient reported way in measuring in situ biological production of superoxide radicals by direct oxidation on non-enzymatic electrodes were based on building carbon microfibers [8,[40][41][42][43][44] with micrometric dimensions. The carbon fiber is sealed in a glass capillary by heating the glass to melt it around the fibre [40] or by using epoxy resin [8,41,42,44].…”

Section: Non-enzymatic Amperometric Sensorsmentioning

confidence: 99%

See 1 more Smart Citation

Selective and sensitive electrochemical biosensing of superoxide anion production by biological systems : a short overview of recent trends

Pontié¹,

Bedioui²

1999

Analusis

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Direct measurement of oscillatory generation of superoxide anions by single phagocytes

Cited by 15 publications

References 15 publications

Salmonella enterica Serovar Typhimurium NiFe Uptake-Type Hydrogenases Are Differentially Expressed In Vivo

Salmonella enterica Serovar Typhimurium NiFe Uptake-Type Hydrogenases Are Differentially Expressed In Vivo

Nitrogen monoxide and oxidative stress: composition and intensity of cellular oxidative bursts cocktail. A study through artificial electrochemical synapses on single human fibroblasts

Selective and sensitive electrochemical biosensing of superoxide anion production by biological systems : a short overview of recent trends

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