Exploring real-time in vivo redox biology of developing and aging Caenorhabditis elegans.
Reactive oxygen species (ROS) are no longer considered merely toxic by-products of the oxidative metabolism. Tightly controlled concentrations of ROS and fluctuations in redox potential may be important mediators of signaling processes. Understanding the role of ROS and redox status in physiology, stress response, development, and aging requires their nondisruptive, spatiotemporal, real-time quantification in a living organism. We established Caenorhabditis elegans strains bearing the genetically encoded fluorescent biosensors HyPer and Grx1-roGFP2 for the detection of hydrogen peroxide (H(2)O(2)) and the glutathione redox potential, respectively. Although, given its transparency and genetic tractability, C. elegans is perfectly suitable as a model organism for such approaches, they have never been tried before in this nematode. We found that H(2)O(2) treatment clearly induces a dose-dependent, reversible response of both biosensors in the living worms. The ratio of oxidized to reduced glutathione decreases during postembryonic development. H(2)O(2) levels increase with age and this effect is delayed when life span is extended by dietary restriction. In young adults, we detected several regions with distinct redox properties that may be linked to their biological function. Our findings demonstrate that genetically encoded biosensors can reveal previously unknown details of in vivo redox biology in multicellular organisms.
Laboratory for Aging Physiology and Molecular Evolution, Department of Biology, Ghent University, Ghent, Belgium.
Patricia
P
Back
Winnok H
WH
De Vos
Geert G
GG
Depuydt
Filip
F
Matthijssens
Jacques R
JR
Vanfleteren
Bart P
BP
Braeckman
D000367:Age Factors; D000818:Animals; D030801:Animals, Genetically Modified; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000378:metabolism; D015374:Biosensing Techniques; D017173:Caenorhabditis elegans / Q000254:growth & development / Q000378:metabolism*; D005786:Gene Expression Regulation; D005835:Genitalia / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D019803:Glutathione Disulfide / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000378:metabolism; D006257:Head; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology*; D008017:Life Expectancy; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000378:metabolism; D009132:Muscles / Q000378:metabolism; D009928:Organ Specificity; D016877:Oxidants / Q000378:metabolism / Q000494:pharmacology*; D010084:Oxidation-Reduction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D013623:Tail / Q000378:metabolism
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D016877:Oxidants; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D049452:Green Fluorescent Proteins; D006861:Hydrogen Peroxide; D005978:Glutathione; D019803:Glutathione Disulfide