Synchronous Growth and Aging of Caenorhabditis elegans in the Presence of Fluorodeoxyuridine

Abstract

Fluorodeoxyuridine (FUdR), an inhibitor of DNA synthesis, was examined for its ability to prevent a synchronous population of C. elegans from reproducing without otherwise interfering with the organism's post-maturational development and aging. When a synchronized population was exposed to 400 micrometer FUdR just as the population reached sexual maturity, the FUdR induced complete sterility within five hours by preventing eggs from hatching. Any larvae that hatched from eggs made before the FUdR was added remained small in the presence of FUdR and were easily removed by filtration or sedimentation. FUdR-sterilized adults showed no morphological abnormalities. Age-associated changes seen in controls also occurred in FUdR-treated worms, including atrophy of the gonads, increased pigmentation, sluggishness and increased transparency. Life span was not shortened by FUdR treatment. Our observations suggest that treatment with FUdR under carefully controlled conditions is a reasonable way to maintain synchronously aging populations of C. elegans.

Mutation of C. elegans demethylase spr-5 extends transgenerational longevity

“…We first treated worms with 5-fluorodeoxyuridine (FUdR), which inhibits proliferation of germline stem cells, the production of intact eggs in adults, and extends longevity [13,14]. We found FUdR had similar effects on the lifespan of WT and early generation spr-5(by101) mutant worms (G5) (Supplementary information, Figure S2A).…”

“…In this paper mutant worms were backcrossed: jmjd-2(tm2966): 2 times, daf-12(m20): 3 times, rde-1(ne219): 3 times, set-17(n5017): 2-7 times, set-30(gk315): 2-8 times, damt-1(gk961032): 5 times, ergo-1(tm1860): 7 times, and eap-1(ok3432): 2-13 times. Generation 1 spr-5 mutant worms were obtained by crossing later generation (generation [10][11][12][13][14][15][16][17][18][19][20] homozygous mutant worms with WT males to obtain P0 heterozygous mutants. Individual P0 heterozygous mutants were picked to plates and allowed to lay generation 1 progeny after which the P0 heterozygous mutant genotype was confirmed by single worm PCR genotyping.…”

Comparative Genomics RNAi Screen Identifies Eftud2 as a Novel Regulator of Innate Immunity

“…The RNAi hypersensitive strain GR1373 eri-1(mg366) IV (Kennedy et al 2004) was synchronized by collecting hatchlings at 15° (Wood 1988), the hatchlings were moved to E. coli-expressing dsRNA and incubated at 25°for 24 hr until they were in the late L4 stage, and then the nematodes were exposed to either PA14 or OP50 at 25°in the presence of 5 mg/ml of the sterilizing agent 5-fluoro-2-deoxyuridine (Mitchell et al 1979) on modified NGM plates (Powell and Ausubel 2008). In the heat-killed bacteria experiment, a liquid OP50 culture was concentrated fivefold, incubated at 80°for 3 hr, and plated on modified NGM supplemented with kanamycin.…”

Subcellular in vivo time-lapse imaging and optical manipulation of Caenorhabditis elegans in standard multiwell plates

“…Each well of a 96-well plate has sufficient volume of media to ensure that C. elegans is healthy over its entire lifespan, and 96-well plates were previously used to enable large-scale screening of C. elegans from L1 stage until death to identify chemicals that extend lifespan 21 . As C. elegans can produce significant numbers of progeny inside the wells, fluorodeoxyuridine is commonly used to induce sterility while still allowing animals to develop normally [21][22][23] . Absence of progeny simplifies tracking of animals throughout their entire lifespan, and reduces both the amount of feeding bacteria required and the buildup of waste inside the wells.…”