Induction of DNA strand breaks by genotoxicants in the alga Chlamydomonas reinhardtii.
The use of unicellular algae in ecotoxicity testing is well established, particularly regarding whole-organism and population-level end points such as lethality and population growth. Conflicting information exists, however, on the potential for genetic toxicity to be incorporated into the safety studies in this test organism. In the present study, DNA strand breaks (Comet assay) and ethoxyresorufin-O-deethylase (EROD) activity were used as indicators of genetic toxicity and cytochrome P450 1A baseline xenobiotic metabolism, respectively, in the unicellular green alga Chlamydomonas reinhardtii. DNA strand breaks were quantified following exposure to the direct-acting genotoxic agents 4-nitroquinoline-1-oxide (NQO) and the N-hydroxy metabolite of 2-acetylaminofluorene (N-OH-2-AAF) and the indirect-acting genotoxin chrysoidine. Following compound exposure, chrysoidine and N-OH-2-AAF produced statistically significant increases in DNA strand breaks at both 0.1 and 10 microM and 0.05 and 5 microM, respectively (p < 0.05 and p < 0.01). Different light sources were also found to influence DNA strand breaks, the minimum response being observed using a source that omits the ultraviolet range. Compared to many mammalian cells, both DNA damage responses and EROD activity were relatively weak. EROD activity was 0.03 pmol/min/10(6) cells in control cells, and the maximum level of DNA strand breaks observed was 14.1% at a 5 nM concentration of NQO. The responses exhibited were not enhanced by the use of a cell wall-free mutant strain. In conclusion, C reinhardtii responded, albeit weakly, to selected direct- and indirect-acting genotoxicants and also exhibited measurable EROD activity.
David
Rhiannon M
RM
School of Biosciences, University of Birmingham, Edgbaston, Birmingham, United Kingdom. r.m.david@bham.ac.uk
Winter
Matthew J
MJ
Chipman
J Kevin
JK
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity; D016825:Chlamydomonas reinhardtii / Q000187:drug effects*; D020552:Comet Assay; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D053960:DNA Breaks; D005978:Glutathione / Q000032:analysis; D009153:Mutagens / Q000633:toxicity*; D010128:p-Aminoazobenzene / Q000031:analogs & derivatives / Q000633:toxicity
D009153:Mutagens; C005562:chrysoidine; D015112:4-Nitroquinoline-1-oxide; D010128:p-Aminoazobenzene; D019363:Cytochrome P-450 CYP1A1; D005978:Glutathione