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  • Transcriptional responses in neonate and adult Daphnia magna in relation to relative susceptibility to genotoxicants.

Transcriptional responses in neonate and adult Daphnia magna in relation to relative susceptibility to genotoxicants.

Aquatic toxicology (Amsterdam, Netherlands) (2011-06-03)
Rhiannon M David, Vanja Dakic, Timothy D Williams, Matthew J Winter, J Kevin Chipman
ABSTRACT

Little information is available on the responses of lower animals to genotoxic chemicals or on their sensitivity for detecting genotoxic chemicals, especially at different life-stages, despite the established use of the water flea Daphnia magna in ecotoxicity testing. Comet assay methodology was developed and applied to daphnid cells but only limited, non-statistically significant responses to the genotoxicants sodium dichromate (0.2-1 μM), chrysoidine (0.1-2 μM), and mixtures of benzo-a-pyrene (BaP) and sodium dichromate were found (from 0.01 μM BaP & 0.1 μM sodium dichromate to 0.25 μM BaP & 0.75 μM sodium dichromate). Transcriptomic analyses using Agilent D. magna oligonucleotide microarrays were undertaken to assess the effect of a mixture of sodium dichromate and BaP (designed to produce both adducted and oxidised DNA) on gene transcription. Neonates (<24h) and adults (day 7) were exposed for 6h and 24h at two combination concentration levels (0.02 μM BaP & 0.15 μM sodium dichromate and 0.1 μM BaP & 0.75 μM sodium dichromate). The greatest differences in transcriptional profile occurred between adults and neonates. Subsets of the transcriptional profiles distinguished genotoxicant-exposed animals from controls, both for neonates and adults. Higher transcript levels of DNA repair genes were found in adults and adults also displayed significant induction of DNA repair gene transcripts in response to exposure whereas neonates did not. Transcriptional changes in response to genotoxicant exposure proved more sensitive than measurement of DNA strand breaks by the Comet assay and the extensive differences in transcription between adults and neonates emphasized the importance of life stage in toxicant testing with Daphnia.

MATERIALS
Product Number
Brand
Product Description

Supelco
4-Aminoazobenzene, analytical standard
Sigma-Aldrich
Sodium dichromate dihydrate, ACS reagent, ≥99.5%
Sigma-Aldrich
Chrysoidine G, for microscopy (Bact., Bot., Vit.)
Supelco
Chrysoidine G, analytical standard