title: Oxidative DNA damage may not mediate Ni-induced genotoxicity in marine mussels: assessment of genotoxic biomarkers and transcriptional responses of key stress genes.
abstract: Nickel (Ni) is a known carcinogenic and mutagenic compound and an important contaminant of aquatic environments. Ni toxicity and its potential impact on aquatic organisms are, however, not well understood. This study used an integrated approach to evaluate genotoxic effects, tissue-specific accumulation and transcriptional profiles of key genes in mussels, Mytilus galloprovincialis, exposed to a range of concentrations of Ni. The genotoxic effects assessed were total and oxidative DNA damage (DNA strand breaks measured using the enzyme modified comet assay), and induction of micronuclei (MN; clastogenic and/or aneugenic effects) using haemocytes as the target cells. Six genes (pgp, mt10, mt20, sod, hsp70 and gst) were selected for transcriptional analysis in the gills based on their key role in the stress response. Following exposure to sublethal concentrations of Ni (0-3600μgL(-1)) for 5 days, mussel haemocytes showed significant genotoxicity at >1800μgL(-1) (4-fold increase for DNA strand breaks and 3-fold increase for MN induction). There was no significant difference between buffer (control) and enzyme treatments which target oxidised DNA bases (formamidopyrimidine glycosylase or endonuclease IIII). This suggested that, in haemocytes, oxidative DNA damage is not a major mechanism for Ni-induced genotoxicity. The expression of mt20 and gst genes in gill was up-regulated at genotoxic concentrations, whilst pgp expression was markedly up-regulated, particularly at 18μgL(-1) Ni (19-fold increase). Pearson's correlation analysis revealed significant associations between % tail DNA and MN induction in haemocytes (r=0.88, p<0.05), and between Ni accumulation in foot (r=0.47, p<0.05) and digestive gland (r=0.41, p<0.05) and induction of MN in the haemocytes. Our results are the first to suggest that Ni-induced genotoxicity in mussel haemocytes may not be a result of oxidative DNA damage, and that multixenobiotic resistance (MXR) may play an important role in Ni detoxification in this species.
authors.affiliation: School of Biomedical & Biological Sciences, Plymouth University, Drake Circus, Plymouth PL4 8AA, UK.
authors.forename: Lorna J
authors.initials: LJ
authors.lastname: Dallas
authors.forename: Tim P
authors.initials: TP
authors.lastname: Bean
authors.forename: Andrew
authors.initials: A
authors.lastname: Turner
authors.forename: Brett P
authors.initials: BP
authors.lastname: Lyons
authors.forename: Awadhesh N
authors.initials: AN
authors.lastname: Jha
mesh_terms: D000818:Animals; D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism*; D049872:Bivalvia / Q000187:drug effects* / Q000235:genetics; D020552:Comet Assay; D004249:DNA Damage; D017931:DNA Primers; D013058:Mass Spectrometry; D009532:Nickel / Q000633:toxicity*; D018384:Oxidative Stress; D060888:Real-Time Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: D015415:Biomarkers; D017931:DNA Primers; D009532:Nickel