An ultrasensitive fluorescent assay for the in vivo quantification of superoxide radical in organisms

Georgiou, Christos D.; Papapostolou, Ioannis; Patsoukis, Nikolaos; Tsegenidis, T.; Sideris, Theodore D.

doi:10.1016/j.ab.2005.09.013

Cited by 69 publications

(74 citation statements)

References 29 publications

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“…The assay is based on the 1:1 molar stoichiometric reaction of O 2 -with DHE that results in the formation of the specific product 2-OH-ethidium the formation rate of which is measured and converted to O 2 -production rate [1]. 2-OHEthidium was estimated after being extracted from the tissue in alkaline acetone, isolated via cation and hydrophobic microcolumn chromatographies and quantified by the use of its fluorescence properties and its destruction by H 2 O 2 (catalyzed by horseradish peroxidase).…”

Section: Superoxide Radical Assaymentioning

confidence: 99%

“…Fluorescence measurements were performed in a quartz microcuvette (internal dimensions 4 9 4 9 45 mm 3 ) with its appropriate holder and a Shimadzu RF-1501 spectrofluorometer set at 10 nm excitation/emission slit width and high sensitivity. The concentration of 2-OH-ethidium was quantified by its standard curve constructed from a stock solution made by the X/XO superoxide generating system in the presence of certain concentration of DHE [1]. The concentration of 2-OH-ethidium in this stock solution was equal to the concentration of consumed DHE since 2-OH-ethidium is the sole product of the reaction of superoxide with DHE.…”

Section: Superoxide Radical Assaymentioning

confidence: 99%

“…1), and the 2-OH-ethidium formed in vivo can be quantified after its isolation by the O 2 -assay [1]. In order to calculate the production rate of O 2 -, the stoichiometric molar ratio 1:1 for the rate of superoxide:rate of 2-OH-ethidium (at the rate range 0-70 nM min -1 O 2 -formation) was used [1].…”

Section: Superoxide Radical Assaymentioning

confidence: 99%

“…For ensuring this, several doses of DHE were injected in the control/tested organisms over different incubation intervals and the rate of O 2 -in the tested organs was determined. After running these preliminary experiments, the proper DHE dose was established based on following criteria: (a) 2-OH-E + formation rate is constant at several incubation periods up to 75 min and (b) that unreacted HE is detected in the brain tissue [1].…”

Section: Administration Of Dhe (Superoxide Radical Trap)mentioning

confidence: 99%

“…This can be possible because of the availability of a new ultrasensitive fluorescent assay that allows the direct measurement of O 2 -in whole organisms in vivo [1].…”

Section: Introductionmentioning

confidence: 99%

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Quantification of Superoxide Radical in the Brain of Rats with Experimentally Induced Obstructive Jaundice

et al. 2007

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The study aimed to directly measure in vivo superoxide radical (O*2) a direct indicator of oxidative stress, in the brain of rats with experimentally induced obstructive jaundice by employing a new quantitative ultrasensitive fluorescent assay requiring minimum sample. O*2 anion is specific for dihydroethidine (DHE) and upon reaction gives a characteristic product, namely 2-OH-ethidium. Ten male rats underwent laparotomy and were divided into two groups: I, sham operated and II bile duct ligation. Ten days later, following injection with DHE (a O*2 trap), all animals were killed and samples from cerebral cortex, midbrain and cerebellum were removed for analysis. It was shown that compared to group I, in group II the O*2 was increased by 67% in the cerebral cortex and by 37% in the midbrain as a consequence of experimental obstructive jaundice, while its levels were unaffected in the cerebellum. The data in this experimental obstructive jaundice model imply a region-specific increase of O*2 formation rate, being higher in cerebral cortex, less so in the midbrain and not at all in cerebellum.

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Section: Superoxide Radical Assaymentioning

confidence: 99%

Section: Superoxide Radical Assaymentioning

confidence: 99%

Section: Superoxide Radical Assaymentioning

confidence: 99%

Section: Administration Of Dhe (Superoxide Radical Trap)mentioning

confidence: 99%

“…This can be possible because of the availability of a new ultrasensitive fluorescent assay that allows the direct measurement of O 2 -in whole organisms in vivo [1].…”

Section: Introductionmentioning

confidence: 99%

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Quantification of Superoxide Radical in the Brain of Rats with Experimentally Induced Obstructive Jaundice

et al. 2007

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2010

Handbook of Biological Dyes and Stains

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Sensors, Imaging Agents, and Theranostics to Help Understand and Treat Reactive Oxygen Species Related Diseases

et al. 2019

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Reactive oxygen species (ROS) consist of a diverse range of oxidative small molecular ions and free radicals that are produced throughout the body during certain biological processes. Due to their high reactivity, these molecules result in the damage of tissues and cells. Therefore, ROS have been implicated in an array of diseases including cancer, inflammation, and neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease. Owing to the simplicity, sensitivity, and selectivity of fluorescence‐based strategies, many small‐molecular sensors or imaging agents have been developed to sense and visualize ROS both in vitro and in vivo. Likewise, activatable drug delivery and prodrug systems that can be triggered by ROS for disease theranostics (diagnostic and therapeutic combined) have been developed. Herein, recently developed fluorescence‐based sensing and imaging agents for the detection of ROS both intracellularly and in vivo are summarized. In addition, drug delivery, which require activation by ROS to achieve disease theranostics is also discussed.

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An ultrasensitive fluorescent assay for the in vivo quantification of superoxide radical in organisms

Cited by 69 publications

References 29 publications

Quantification of Superoxide Radical in the Brain of Rats with Experimentally Induced Obstructive Jaundice

Quantification of Superoxide Radical in the Brain of Rats with Experimentally Induced Obstructive Jaundice

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Sensors, Imaging Agents, and Theranostics to Help Understand and Treat Reactive Oxygen Species Related Diseases

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