Measuring Oxidative Stress in Caenorhabditis elegans: Paraquat and Juglone Sensitivity Assays

Senchuk, Megan M.; Dues, Dylan J.; Raamsdonk, Jeremy M. Van

doi:10.21769/bioprotoc.2086

Cited by 58 publications

(42 citation statements)

References 10 publications

(16 reference statements)

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“…We utilized four different stresses: oxidative stress through exposure to 2 mM paraquat for 24 hours, heat stress through exposure to 37°C heat for 2 hours, osmotic stress through exposure to 400 mM NaCl for 24 hours, and complete anoxia for 24 hours. These specific stress paradigms have been previously utilized by our laboratory as well as several others 38‐44 . In this experiment, we chose to expose worms to a shorter duration or smaller dose of stress than we would use in a test of stress resistance to ensure that all worms would survive, and so that we could observe the effect of stress, rather than dying, on mitochondrial morphology.…”

Section: Resultsmentioning

confidence: 99%

Disruption of mitochondrial dynamics increases stress resistance through activation of multiple stress response pathways

et al. 2020

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Mitochondria are dynamic organelles that can change shape and size depending on the needs of the cell through the processes of mitochondrial fission and fusion. In this work, we investigated the role of mitochondrial dynamics in organismal stress response. By using C. elegans as a genetic model, we could visualize mitochondrial morphology in a live organism with well‐established stress assays and well‐characterized stress response pathways. We found that disrupting mitochondrial fission (DRP1/drp‐1) or fusion (OPA1/eat‐3, MFN/fzo‐1) genes caused alterations in mitochondrial morphology that impacted both mitochondrial function and physiologic rates. While both mitochondrial fission and mitochondrial fusion mutants showed increased sensitivity to osmotic stress and anoxia, surprisingly we found that the mitochondrial fusion mutants eat‐3 and fzo‐1 are more resistant to both heat stress and oxidative stress. In exploring the mechanism of increased stress resistance, we found that disruption of mitochondrial fusion genes resulted in the upregulation of multiple stress response pathways. Overall, this work demonstrates that disrupting mitochondrial dynamics can have opposite effects on resistance to different types of stress. Our results suggest that disruption of mitochondrial fusion activates multiple stress response pathways that enhance resistance to specific stresses.

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Section: Resultsmentioning

confidence: 99%

Disruption of mitochondrial dynamics increases stress resistance through activation of multiple stress response pathways

et al. 2020

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“…It has been suggested that microbial infections induce oxidative stress in C. elegans (Bolm et al, 2004;Miller et al, 2015;Sem and Rhen, 2012). We therefore treated worms with paraquat that is commonly used as an oxidative stress inducer (Senchuk et al, 2017). Fig.…”

Section: Endotube Dysfunction Correlates With Impaired Oxidative and mentioning

confidence: 99%

“…Methyl viologen dichloride hydrate ( paraquat; Sigma-Aldrich) was used for oxidative stress assays at a final concentration of 200 mM in NGM agar (Senchuk et al, 2017). Poured plates were stored overnight at room temperature following inoculation with 50 µl 10× concentrated overnight culture of OP50.…”

Section: Stress Assaysmentioning

confidence: 99%

The intestinal intermediate filament network responds to and protects against microbial insults and toxins

et al. 2019

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The enrichment of intermediate filaments in the apical cytoplasm of intestinal cells is evolutionarily conserved, forming a sheath that is anchored to apical junctions and positioned below the microvillar brush border, which suggests a protective intracellular barrier function. To test this, we used Caenorhabditis elegans, the intestinal cells of which are endowed with a particularly dense intermediate filament-rich layer that is referred to as the endotube. We found alterations in endotube structure and intermediate filament expression upon infection with nematicidal B. thuringiensis or treatment with its major pore-forming toxin crystal protein Cry5B. Endotube impairment due to defined genetic mutations of intermediate filaments and their regulators results in increased Cry5B sensitivity as evidenced by elevated larval arrest, prolonged time of larval development and reduced survival. Phenotype severity reflects the extent of endotube alterations and correlates with reduced rescue upon toxin removal. The results provide in vivo evidence for a major protective role of a properly configured intermediate filament network as an intracellular barrier in intestinal cells. This notion is further supported by increased sensitivity of endotube mutants to oxidative and osmotic stress.

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“…[5]. In addition, oxidative stress has been proposed to be one of the major causes of aging and has been implicated in the pathogenesis of many diseases [8]. Therefore, a certain concentration of antioxidants will contribute to the longevity of organisms.…”

Section: Introductionmentioning

confidence: 99%

Hydrogen extends Caenorhabditis elegans longevity by reducing reactive oxygen species

Zhang

Gao

et al. 2020

PLoS ONE

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At present, a large number of studies have reported that hydrogen has antioxidant functions and prevents oxidative stress damage. However, it is not clear whether hydrogen can prolong longevity based on these effects. Therefore, we studied and explored the antiaging potential of exogenous hydrogen and its ability to extend longevity using Caenorhabditis elegans (C. elegans) as an animal model. Our results showed that the lifespans of the N2, sod-3 and sod-5 mutant strains were extended by approximately 22.7%, 9.5%, and 8.7%, respectively, after hydrogen treatment, but hydrogen had no effect on the lifespans of the daf-2 and daf-16 mutant strains. Meanwhile, the level of reactive oxygen species (ROS) in the hydrogen treatment group was significantly lower than that in the control group. At the transcript level, the expression of age-1 and let-363 was obviously decreased, while the expression of ins-18 was increased at the same time point (14 d). Compared with the control group, paraquat (PQ) could reduce the lifespan of the N2 and sod-5 mutant strains. Importantly, the longevity of these mutant strains recovered to normal levels when the animals were treated with exogenous hydrogen. According to these results, the lifespan of C. elegans is closely related to oxidative stress and can be significantly prolonged by reducing oxidative stress damage. Taken together, our data showed that hydrogen is a valuable antioxidant that can significantly reduce the body's ROS levels and extend the lifespan of C. elegans. This study also laid a foundation for the subsequent application of hydrogen in antiaging studies.

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Measuring Oxidative Stress in Caenorhabditis elegans: Paraquat and Juglone Sensitivity Assays

Cited by 58 publications

References 10 publications

Disruption of mitochondrial dynamics increases stress resistance through activation of multiple stress response pathways

Disruption of mitochondrial dynamics increases stress resistance through activation of multiple stress response pathways

The intestinal intermediate filament network responds to and protects against microbial insults and toxins

Hydrogen extends Caenorhabditis elegans longevity by reducing reactive oxygen species

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