Oxidative Stress Assays (arsenite and tBHP) in Caenorhabditis elegans

Ewald, Collin Y.; Hourihan, John M.; Blackwell, T. Keith

doi:10.21769/bioprotoc.2365

Cited by 17 publications

(13 citation statements)

References 11 publications

(18 reference statements)

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“…The log-rank (Mantel-Cox) method was used to test the null hypothesis and calculate P values (JMP software v.9.0.2.). Both oxidative stress tolerance assays are described in more detail at Bio-protocol (Ewald et al, 2017). …”

Section: Methodsmentioning

confidence: 99%

NADPH oxidase-mediated redox signaling promotes oxidative stress resistance and longevity through memo-1 in C. elegans

Ewald

Hourihan

Bland

et al. 2017

eLife

103

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Transient increases in mitochondrially-derived reactive oxygen species (ROS) activate an adaptive stress response to promote longevity. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidases produce ROS locally in response to various stimuli, and thereby regulate many cellular processes, but their role in aging remains unexplored. Here, we identified the C. elegans orthologue of mammalian mediator of ErbB2-driven cell motility, MEMO-1, as a protein that inhibits BLI-3/NADPH oxidase. MEMO-1 is complexed with RHO-1/RhoA/GTPase and loss of memo-1 results in an enhanced interaction of RHO-1 with BLI-3/NADPH oxidase, thereby stimulating ROS production that signal via p38 MAP kinase to the transcription factor SKN-1/NRF1,2,3 to promote stress resistance and longevity. Either loss of memo-1 or increasing BLI-3/NADPH oxidase activity by overexpression is sufficient to increase lifespan. Together, these findings demonstrate that NADPH oxidase-induced redox signaling initiates a transcriptional response that protects the cell and organism, and can promote both stress resistance and longevity.DOI: http://dx.doi.org/10.7554/eLife.19493.001

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Section: Methodsmentioning

confidence: 99%

NADPH oxidase-mediated redox signaling promotes oxidative stress resistance and longevity through memo-1 in C. elegans

Ewald

Hourihan

Bland

et al. 2017

eLife

103

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“…Oxidative stress assay was modified from Ewald et al, 2017(Ewald et al, 2017. C. elegans of the L1 or day-1-adult stage were shifted on compound-containing or control plates washed off at the indicated time point, incubated with 14 mM sodium arsenite (Honeywell International 35000) in U-Shaped 96 well plates, and put into the wMicroTracker (MTK100) for movement scoring.…”

Section: Oxidative Stress Assaysmentioning

confidence: 99%

Youthful and age-related matreotypes predict drugs promoting longevity

Statzer

Jongsma

Liu

et al. 2021

Preprint

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The identification and validation of drugs that promote health during aging (‘geroprotectors’) is key to the retardation or prevention of chronic age-related diseases. Here we found that most of the established pro-longevity compounds shown to extend lifespan in model organisms also alter extracellular matrix gene expression (i.e., matrisome) in human cell lines. To harness this novel observation, we used age-stratified human transcriptomes to define the age-related matreotype, which represents the matrisome gene expression pattern associated with age. Using a ‘youthful’ matreotype, we screened in silico for geroprotective drug candidates. To validate drug candidates, we developed a novel tool using prolonged collagen expression as a non-invasive and in-vivo surrogate marker for C. elegans longevity. With this reporter, we were able to eliminate false positive drug candidates and determine the appropriate dose for extending the lifespan of C. elegans. We improved drug uptake for one of our predicted compounds, genistein, and reconciled previous contradictory reports of its effects on longevity. We identified and validated new compounds, tretinoin, chondroitin sulfate, and hyaluronic acid, for their ability to restore age-related decline of collagen homeostasis and increase lifespan. Thus, our innovative drug screening approach - employing extracellular matrix homeostasis - facilitates the discovery of pharmacological interventions promoting healthy aging.HighlightsMany geroprotective drugs alter extracellular matrix gene expressionDefined young and old human matreotype signatures can identify novel potential geroprotective compoundsProlonged collagen homeostasis as a surrogate marker for longevity

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“…How NSY-1 becomes activated upon oxidative insult is unclear. However, the inorganic compound sodium arsenite is known to induce ROS and activate the p38MAPK/Nrf2 oxidative stress response in mammals and C. elegans [ 34 , 68 , 69 , 70 , 71 , 72 ]. Treating C. elegans with sodium arsenite leads to a physical interaction between BLI-3/Duox and endoplasmic reticulum membrane-bound kinase IRE-1 [ 34 ].…”

Section: Bli-3-generated Ros As a Signaling Molecule To Activate Tmentioning

confidence: 99%

Redox Signaling of NADPH Oxidases Regulates Oxidative Stress Responses, Immunity and Aging

Ewald

2018

Antioxidants

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An accumulating body of evidence suggests that transient or physiological reactive oxygen species (ROS) generated by nicotinamide adenine dinucleotide phosphate (NADPH) oxidases act as a redox signal to re-establish homeostasis. The capacity to re-establish homeostasis progressively declines during aging but is maintained in long-lived animals to promote healthy aging. In the model organism Caenorhabditis elegans, ROS generated by dual oxidases (Duox) are important for extracellular matrix integrity, pathogen defense, oxidative stress resistance, and longevity. The Duox enzymatic activity is tightly regulated and under cellular control. Developmental molting cycles, pathogen infections, toxins, mitochondrial-derived ROS, drugs, and small GTPases (e.g., RHO-1) can activate Duox (BLI-3) to generate ROS, whereas NADPH oxidase inhibitors and negative regulators, such as MEMO-1, can inhibit Duox from generating ROS. Three mechanisms-of-action have been discovered for the Duox/BLI-3-generated ROS: (1) enzymatic activity to catalyze crosslinking of free tyrosine ethyl ester in collagen bundles to stabilize extracellular matrices, (2) high ROS bursts/levels to kill pathogens, and (3) redox signaling activating downstream kinase cascades to transcription factors orchestrating oxidative stress and immunity responses to re-establish homeostasis. Although Duox function at the cell surface is well established, recent genetic and biochemical data also suggests a novel role for Duoxs at the endoplasmic reticulum membrane to control redox signaling. Evidence underlying these mechanisms initiated by ROS from NADPH oxidases, and their relevance for human aging, are discussed in this review. Appropriately controlling NADPH oxidase activity for local and physiological redox signaling to maintain cellular homeostasis might be a therapeutic strategy to promote healthy aging.

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Oxidative Stress Assays (arsenite and tBHP) in Caenorhabditis elegans

Cited by 17 publications

References 11 publications

NADPH oxidase-mediated redox signaling promotes oxidative stress resistance and longevity through memo-1 in C. elegans

NADPH oxidase-mediated redox signaling promotes oxidative stress resistance and longevity through memo-1 in C. elegans

Youthful and age-related matreotypes predict drugs promoting longevity

Redox Signaling of NADPH Oxidases Regulates Oxidative Stress Responses, Immunity and Aging

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