Effects of 5'-fluoro-2-deoxyuridine on mitochondrial biology in Caenorhabditis elegans.
5-Fluoro-2'-deoxyuridine (FUdR) is a DNA synthesis inhibitor commonly used to sterilize Caenorhabditis elegans in order to maintain a synchronized aging population of nematodes, without contamination by their progeny, in lifespan experiments. All somatic cells in the adult nematode are post-mitotic and therefore do not require nuclear DNA synthesis. However, mitochondrial DNA (mtDNA) replicates independently of the cell cycle and thus represents a potential target for FUdR toxicity. Inhibition of mtDNA synthesis can lead to mtDNA depletion, which is linked to a number of diseases in humans. Furthermore, alterations in mitochondrial biology can affect lifespan in C. elegans. We characterized the effects of FUdR exposure on mtDNA and nuclear DNA (nucDNA) copy numbers, DNA damage, steady state ATP levels, nematode size, mitochondrial morphology, and lifespan in the germ line deficient JK1107 glp-1(q244) and PE255 glp-4(bn2) strains. Lifespan was increased very slightly by 25 μM FUdR, but was reduced by 400 μM. Both concentrations reduced mtDNA and nucDNA copy numbers, but did not change their ratio. There was no detectable effect of FUdR on mitochondrial morphology. Although both concentrations of FUdR resulted in smaller sized animals, changes to steady-state ATP levels were either not detected or restricted to the higher dose and/or later timepoints, depending on the method employed and strain tested. Finally, we determined the half-life of mtDNA in somatic cells of adult C. elegans to be between 8 and 13 days; this long half-life very likely explains the small or undetectable impact of FUdR on mitochondrial endpoints in our experiments. We discuss the relative pitfalls associated with using FUdR and germline deficient mutant strains as tools for the experimental elimination of progeny.
Duke University, Nicholas School of the Environment, Integrated Toxicology and Environmental Health Program, LSRC, PO Box 90328, Durham, NC 27708, USA.
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Rooney
Duke University, Nicholas School of the Environment, Integrated Toxicology and Environmental Health Program, LSRC, PO Box 90328, Durham, NC 27708, USA.
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Luz
Duke University, Nicholas School of the Environment, Integrated Toxicology and Environmental Health Program, LSRC, PO Box 90328, Durham, NC 27708, USA.
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González-Hunt
Duke University, Nicholas School of the Environment, Integrated Toxicology and Environmental Health Program, LSRC, PO Box 90328, Durham, NC 27708, USA.
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Bodhicharla
Duke University, Nicholas School of the Environment, Integrated Toxicology and Environmental Health Program, LSRC, PO Box 90328, Durham, NC 27708, USA.
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Ryde
Duke University, Nicholas School of the Environment, Integrated Toxicology and Environmental Health Program, LSRC, PO Box 90328, Durham, NC 27708, USA.
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Anbalagan
Duke University, Nicholas School of the Environment, Integrated Toxicology and Environmental Health Program, LSRC, PO Box 90328, Durham, NC 27708, USA. Electronic address: joel.meyer@duke.edu.
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Meyer
D000255:Adenosine Triphosphate / Q000378:metabolism; D000375:Aging / Q000378:metabolism; D000818:Animals; D017173:Caenorhabditis elegans / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D004249:DNA Damage; D004261:DNA Replication / Q000187:drug effects*; D004272:DNA, Mitochondrial / Q000096:biosynthesis / Q000187:drug effects* / Q000378:metabolism; D003857:Deoxyuridine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004734:Energy Metabolism / Q000187:drug effects; D018628:Gene Dosage; D005838:Genotype; D006207:Half-Life; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D010641:Phenotype; D013997:Time Factors
C576827:5-fluoro-2'-deoxyuridine; D004272:DNA, Mitochondrial; D000255:Adenosine Triphosphate; D003857:Deoxyuridine
Aging; Caenorhabditis elegans; Copy number; FUdR; Lifespan; Mitochondrial DNA; Mitochondrial DNA half-life; Mitochondrial toxicity