A
 C. elegans
 Screening Platform for the Rapid Assessment of Chemical Disruption of Germline Function

Allard, Patrick; Kleinstreuer, Nicole; Knudsen, Thomas B.; Colaiácovo, Mónica P.

doi:10.1289/ehp.1206301

Cited by 76 publications

(89 citation statements)

References 32 publications

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“…Exposed worms are directly imaged on 384-well plates and the number of worms containing at least one GFP-positive embryo is counted for each well and normalized by the total number of worms in that well. A positive hit from the chemical screen means a compound induced a proportion of GFP-positive embryos in a worm population at a frequency 1.7× higher than DMSO 10 . Following threshold optimization, the high content image analysis (see Materials file) allows the automated calculation of the number of positive objects ( i.e ., embryos) divided by the total number of worms in the well and is the method of choice for the large scale adaptation of the assay.…”

Section: Representative Resultsmentioning

confidence: 99%

“…Adapted to chemical screening, this strain is employed in a medium to high-throughput screen. Importantly, the strain faithfully reports the aneugenicity of chemicals and is therefore relevant to mammalian reproductive endpoints 10 . The assay described here will be particularly useful to toxicologists in pharmaceutical and chemical industry settings looking to rapidly assess the toxicity of chemicals towards reproductive endpoints.…”

Section: Introductionmentioning

confidence: 95%

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Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode Caenorhabditis elegans

Parodi

Damoiseaux

Allard

2015

JoVE

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Identifying the reproductive toxicity of the thousands of chemicals present in our environment has been one of the most tantalizing challenges in the field of environmental health. This is due in part to the paucity of model systems that can (1) accurately recapitulate keys features of reproductive processes and (2) do so in a medium- to high-throughput fashion, without the need for a high number of vertebrate animals. We describe here an assay in the nematode C. elegans that allows the rapid identification of germline toxicants by monitoring the induction of aneuploid embryos. By making use of a GFP reporter line, errors in chromosome segregation resulting from germline disruption are easily visualized and quantified by automated fluorescence microscopy. Thus the screening of a particular set of compounds for its toxicity can be performed in a 96- to 384-well plate format in a matter of days. Secondary analysis of positive hits can be performed to determine whether the chromosome abnormalities originated from meiotic disruption or from early embryonic chromosome segregation errors. Altogether, this assay represents a fast first-pass strategy for the rapid assessment of germline dysfunction following chemical exposure.

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Section: Representative Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 95%

Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode Caenorhabditis elegans

Parodi

Damoiseaux

Allard

2015

JoVE

Self Cite

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“…Although the assay is in its infancy and will require further validation, preliminary analysis of a selection of 50 chemicals from ToxCast phase 1 and known chemicals revealed a maximum balanced accuracy (representing the average of sensitivity and specificity) of 69% in predicting the ability of chemicals that cause reproductive toxicity in rodents [9]. The technology was viewed favorably by the workshop participants, and it was recommended that this assay be considered for integration with HTS assays as part of tier 1 screening.…”

Section: High-throughput Analysis Of Egg Aneuploidy In C Elegansmentioning

confidence: 99%

“…Several practical assays have emerged more recently that address some of the gaps in testing described above: (1) the transgenic rodent mutation assays OECD guideline (TG488) that includes recommendations for male germ cell mutation analysis; (2) sperm and pedigree tandem repeat mutation analysis [7]; (3) improved methods to quantify sperm DNA damage and chromatin effects [8]; and (4) high-throughput screening for aneuploidy in Caenorhabditis elegans eggs [9]. Most importantly, the rapid technological evolution of genomics tools, including DNA microarrays and next generation sequencing, is poised to revolutionize the field dramatically.…”

Section: Introductionmentioning

confidence: 99%

Approaches for identifying germ cell mutagens: Report of the 2013 IWGT workshop on germ cell assays☆

Yauk

Aardema

Benthem

et al. 2015

Mutation Research/Genetic Toxicology and Environmental Mutagene

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This workshop reviewed the current science to inform and recommend the best evidence-based approaches on the use of germ cell genotoxicity tests. The workshop questions and key outcomes were as follows. (1) Do genotoxicity and mutagenicity assays in somatic cells predict germ cell effects? Limited data suggest that somatic cell tests detect most germ cell mutagens, but there are strong concerns that dictate caution in drawing conclusions. (2) Should germ cell tests be done, and when? If there is evidence that a chemical or its metabolite(s) will not reach target germ cells or gonadal tissue, it is not necessary to conduct germ cell tests, notwithstanding somatic outcomes. However, it was recommended that negative somatic cell mutagens with clear evidence for gonadal exposure and evidence of toxicity in germ cells could be considered for germ cell mutagenicity testing. For somatic mutagens that are known to reach the gonadal compartments and expose germ cells, the chemical could be assumed to be a germ cell mutagen without further testing. Nevertheless, germ cell mutagenicity testing would be needed for quantitative risk assessment. (3) What new assays should be implemented and how? There is an immediate need for research on the application of whole genome sequencing in heritable mutation analysis in humans and animals, and integration of germ cell assays with somatic cell genotoxicity tests. Focus should be on environmental exposures that can cause de novo mutations, particularly newly recognized types of genomic changes. Mutational events, which may occur by exposure of germ cells during embryonic development, should also be investigated. Finally, where there are indications of germ cell toxicity in repeat dose or reproductive toxicology tests, consideration should be given to leveraging those studies to inform of possible germ cell genotoxicity.

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“…For example, in order to being able to link HTS cellular toxicity data to molecular targets in multicellular organisms, investigators have measured behavioural responses and motor activity of D. melanogaster to airborne VOCs as a model system for discovering adverse outcome pathways and as a method of toxicity screening (Tatum-Gibbs et al, 2015). Other researchers have developed toxicity platforms using C. elegans to study environmental chemical effects on germline function, induction of aneuploidy and prediction of reproductive deficits (Allard et al, 2013). In related efforts, more high-throughput methods are being developed for assessing chemical toxicity in a C. elegans reproduction assay (Boyd et al, 2010).…”

Section: Tox 21: the Futurementioning

confidence: 99%

Intersection of toxicogenomics and high throughput screening in the Tox21 program: an NIEHS perspective

Merrick

Paules

Tice

2015

IJBT

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Humans are exposed to thousands of chemicals with inadequate toxicological data. Advances in computational toxicology, robotic high throughput screening (HTS), and genome-wide expression have been integrated into the Tox21 program to better predict the toxicological effects of chemicals. Tox21 is a collaboration among US government agencies initiated in 2008 that aims to shift chemical hazard assessment from traditional animal toxicology to target-specific, mechanism-based, biological observations using in vitro assays and lower organism models. HTS uses biocomputational methods for probing thousands of chemicals in in vitro assays for gene-pathway response patterns predictive of adverse human health outcomes. In 1999, NIEHS began exploring the application of toxicogenomics to toxicology and recent advances in NextGen sequencing should greatly enhance the biological content obtained from HTS platforms. We foresee an intersection of new technologies in toxicogenomics and HTS as an innovative development in Tox21. Tox21 goals, priorities, progress, and challenges will be reviewed.

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A C. elegans Screening Platform for the Rapid Assessment of Chemical Disruption of Germline Function

Cited by 76 publications

References 32 publications

Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode <em>Caenorhabditis elegans</em>

Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode <em>Caenorhabditis elegans</em>

Approaches for identifying germ cell mutagens: Report of the 2013 IWGT workshop on germ cell assays☆

Intersection of toxicogenomics and high throughput screening in the Tox21 program: an NIEHS perspective

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