The DOI matches its Crossref record; 18 of the 18 references it cites were found in a registry or an index.
Measurement of Reactive Oxygen Species (ROS) and Mitochondrial ROS in AMPK Knockout Mice Blood Vessels
Open accessAbstract
Reactive oxygen species (ROS) are a group of unstable and highly reactive molecules or free radicals typically generated as by-products of cellular processes involving molecular oxygen. In vascular cells, the excessive ROS generation results in the initiation and progression of cardiovascular diseases (CVD). Therefore, a dynamic, robust, and accurate ROS detection method in the blood vessels is essential for pathophysiological research studies of the cardiovascular system.In this chapter, we describe a fluorescence dye-based detection method for assaying superoxide and mitochondrial superoxide in mouse aorta using dihydroethidium (DHE) and MitoSOX. The protocol includes preparation of frozen aortic tissue sections, monitoring DHE oxidation-derived fluorescence by fluorescence microscopy, and high-performance liquid chromatograph-based analysis of MitoSOX and its oxidation products. For studying the role of AMP-activated protein kinase (AMPK) in the redox regulation, we employed AMPKα2 knockout mice and observed increased superoxide and mitochondrial superoxide levels in the aorta of AMPK knockout mice relative to the wild-type group. This novel ROS detection method will be valuable for investigating the roles of cellular and/or mitochondrial ROS in the pathogenesis of CVDs.
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“Dihydroethidium (DHE) is a red fluorescent probe that rather specifically reacts with cellular O2●−, resulting in the two fluorescent products ethidium and 2-hydroxyethidium. Only 2-hydroxyethidium is generated after reaction with O2●−, while ethidium is formed by nonspecific redox reactions. However, DHE easily crosses cellular membranes and therefore can be oxidized by O2●− anywhere in and”
From Functions of ROS in Macrophages and Antimicrobial Immunity (2021) · cites this paper as [360]
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“MitoSOX Red (MitoSOX) is a modified DHE analog attached to a triphenylphosphonium (TPP) group, which leads to accumulation of the probe in the mitochondrial matrix [365,366]. Like DHE itself, MitoSOX specifically detects O2●− and emits red fluorescence after the reaction (see Figure 7) [30,99,360]. In contrast to DHE, MitoSOX does so mainly in the mitochondrial matrix, though. Notably, MitoSOX can”
From Functions of ROS in Macrophages and Antimicrobial Immunity (2021) · cites this paper as [360]
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Metadata
- Authors
- Wang, Q., & Zou, M.-H.
- Affiliations
- Georgia State University (US)
- Corresponding author
- Zou, Ming-Hui
- Venue
- Methods in Molecular Biology
- Publisher
- Springer New York
- Published
- 2018
- Volume
- 1732
- Pages
- 507-517
- ISSN
- 1064-3745 (ISSN-L)
- 1940-6029
- PMC
- PMC6407612
- OpenAlex
- W2789049383
- Cited by
- 125
- References
- 18
- Citations by year
- 2026: 10 · 2025: 13 · 2024: 15 · 2023: 19 · 2022: 24 · 2021: 19 · 2020: 18 · 2019: 4 · 2018: 2
- FWCI
- 14.44
- Type
- book chapter
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Methods in molecular biology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute on Aging (R01AG047776)
- National Heart, Lung, and Blood Institute (R01HL105157, R01HL079584, R01HL142287, R01HL137371, R01HL132500, R01HL074399, R01HL080499, R01HL140954, R01HL110488, R01HL128014, R01HL089920, R01HL096032)
- National Cancer Institute (R01CA213022)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Health Sciences › Medicine › Physiology (medical) › Nitric Oxide and Endothelin Effects
- Health Sciences › Medicine › Biochemistry (medical) › Antioxidant Activity and Oxidative Stress
- Keywords
- reactive oxygen species, mitochondrial superoxide, superoxide, AMPK, aorta, dihydroethidium, MitoSOX, cardiovascular disease, redox regulation, vascular cells
- MeSH terms
- Animals, Aorta, Cardiovascular Diseases, Chromatography, High Pressure Liquid, Ethidium, Fluorescent Dyes, Microscopy, Fluorescence, Mitochondria, Oxidation-Reduction, Phenanthridines, Reactive Oxygen Species, Mice, Knockout, Myocytes, Smooth Muscle, Endothelial Cells, Mice, AMP-Activated Protein Kinases
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