Reproductive Toxicity

The assay to study the potential effects of chemicals on reproductive capacity of C. elegans.

Method description

A synchronized population of C. elegans is injected into the microfluidic platform at the first larval stage (L1). Worms are then confined within dedicated microfluidic chambers and are continuously fed with an E. coli solution.

Worms are chronically exposed to the test compounds starting from the last larval stage prior to sexual maturity (L4) for 85 hours (day 3 of adulthood). The compounds to be tested are mixed with the E. coli solution. Freeze-dried OP50 E. coli are used as a food source for the whole duration of the experiment, preventing the metabolization of the tested molecules by the bacteria.

The images of each microfluidic chamber are recorded every hour. Time-resolved phenotypic readouts are then extracted from the collected images.

Reproduction timeline

Readouts

  • Sexual maturity
  • Fertility
  • Embryonic viability
  • Progeny accumulation rate
  • Survival rate
  • Worm size
  • Growth dynamics

C. elegans reproduction over 96 hours post-hatching for wild-type N2 worms treated with the anticancer drug 5-fluorouracil starting from the L4 larval stage (right) and untreated worms (left) as observed via brightfield imaging by the SydLab system. Overlay masks for the detection of worms and eggs are automatically generated by SydLab’s AI-based computer vision algorithm.

Progeny production dynamics

Temporal evolution of the average number of larvae produced by wild-type N2 C. elegans treated with the anticancer drug 5-fluorouracil (5-FU) starting from the L4 larval stage vs untreated worms (vehicle). The drug treatment significantly impacts on C. elegans reproductive process.

Reproduction rate

Progeny production rate over the first 24 hours of reproduction for wild-type N2 C. elegans treated with the anticancer drug 5-fluorouracil (5-FU) at different doses (data normalized to the value measured for the untreated worm population). 5-fluorouracil treatments dose-dependently induce an embryotoxic effect.

Assay combinations

The reproductive toxicity assay can be offered on its own or in combination with the following assays upon request.