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. 2010 Jun 1;245(2):153-9.
doi: 10.1016/j.taap.2010.02.014. Epub 2010 Mar 4.

A high-throughput method for assessing chemical toxicity using a Caenorhabditis elegans reproduction assay

Windy A Boyd  1 , Sandra J McBride, Julie R Rice, Daniel W Snyder, Jonathan H Freedman
Affiliations

A high-throughput method for assessing chemical toxicity using a Caenorhabditis elegans reproduction assay

Windy A Boyd et al. Toxicol Appl Pharmacol. .

Abstract

The National Research Council has outlined the need for non-mammalian toxicological models to test the potential health effects of a large number of chemicals while also reducing the use of traditional animal models. The nematode Caenorhabditis elegans is an attractive alternative model because of its well-characterized and evolutionarily conserved biology, low cost, and ability to be used in high-throughput screening. A high-throughput method is described for quantifying the reproductive capacity of C. elegans exposed to chemicals for 48 h from the last larval stage (L4) to adulthood using a COPAS Biosort. Initially, the effects of exposure conditions that could influence reproduction were defined. Concentrations of DMSO vehicle <or=1% did not affect reproduction. Previous studies indicated that C. elegans may be influenced by exposure to low pH conditions. At pHs greater than 4.5, C. elegans reproduction was not affected; however below this pH there was a significant decrease in the number of offspring. Cadmium chloride was chosen as a model toxicant to verify that automated measurements were comparable to those of traditional observational studies. EC(50) values for cadmium for automated measurements (176-192 microM) were comparable to those previously reported for a 72-h exposure using manual counting (151 microM). The toxicity of seven test toxicants on C. elegans reproduction was highly correlative with rodent lethality suggesting that this assay may be useful in predicting the potential toxicity of chemicals in other organisms.

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Figures

Figure 1
Figure 1. Scatter plots illustrating concentration-dependent decreases in C. elegans reproduction
Extinction (log10 scale) plotted against time of flight (log10 scale) for untreated (left panel), 2.5% DMSO (center panel), and 5.5% DMSO (right panel) wild-type nematodes. C. elegans L4s (gray circles) were loaded at t=0 h. After 48 h, the nematodes were recovered as adults and their offspring (black circles). Each point corresponds to a single nematode. (AU, arbitrary units)
Figure 2
Figure 2. Effect of DMSO on C. elegans reproduction
Total number of offspring plotted against DMSO concentration (%) of wild-type (upper panel) and myo-2::GFP (lower panel) strains for one representative experiment. The solid, black vertical lines represent a 50% reduction in the total number of offspring (EC50). The dashed, black vertical lines represent the BMC, below which the reproductive rate is not significantly affected by DMSO.
Figure 3
Figure 3. Effects of pH on C. elegans reproduction
Total number of offspring plotted against pH for one representative experiment. Increasing concentrations of ascorbic acid (upper panel) or acetic acid (lower panel) caused a decrease in the number of wild-type and myo-2::GFP offspring, respectively. The solid, black vertical lines represent a 50% reduction in the total number of offspring (pH50). The dashed, black vertical lines represent the BMpH, above which the reproductive rate is not significantly affected by pH.
Figure 4
Figure 4. Effect of cadmium on C. elegans reproduction
Total number of offspring plotted against cadmium concentration (μM) of wild-type (upper panel) and myo-2::GFP (lower panel) strains for one representative experiment. The solid, black vertical lines represent a 50% reduction in the total number of offspring (EC50). The dashed, black vertical lines represent the BMC, below which the reproductive rate is not significantly affected by cadmium.
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