Cemagref, Unité de Recherche Hydrosystèmes et Bioprocédés, 1 rue Pierre-Gilles de Gennes, CS10030 - 92761 Antony, France.
Mutagenesis. 2012 Nov;27(6):703-11. doi: 10.1093/mutage/ges036. Epub 2012 Jul 27.
The use of DNA adduct analysis has previously focused on the use of marine organisms for biomonitoring, whereas similar investigations in freshwater organisms are sparse. In that context, we have investigated the relevance of DNA adducts as biomarkers of genotoxicity in the freshwater mussels Dreissena polymorpha. The objective of the present study is to determine the stability of DNA adducts induced by benzo[a]pyrene (B[a]P) in zebra mussels. Mussels were exposed to dissolved B[a]P (10-100 µg/l) for 4 days. Afterwards, mussels were kept in clean water for 28 days and DNA adduct levels were subsequently measured in two different organs, the digestive glands and the gills, using the (32)P-postlabelling technique. In parallel, the expression of genes involved in the detoxification system was assessed by qPCR (catalase, superoxide dismutase, glutathione S transferase, HSP70, aryl hydrocarbon receptor, P glycoprotein). We observed a higher level of DNA adducts in the digestive glands compared to the gills. Moreover, in gills, the level of DNA adduct was dependent on the B[a]P concentration. The levels of adducts tended to decrease in both organs after 28 days in clean water. In addition, an early induction of HSP70, PgP, AHR and SOD mRNA levels was noticed in the gills compared to the digestive glands. CAT and GST gene expression increased from 12h exposure in both organs. A higher gene expression level of those genes was observed in the gills, except for AHR and CAT genes. Data converge towards the fact that DNA adducts hence represent a very promising biomarker of B[a]P contamination and potentially of exposure to polycyclic aromatic hydrocarbons. In addition, for the first time in this study, B[a]P detoxification system was characterised in D. polymorpha.
先前的 DNA 加合物分析主要集中在利用海洋生物进行生物监测,而对淡水生物的类似研究则很少。在这种情况下,我们研究了 DNA 加合物作为淡水贻贝多形帘蛤遗传毒性生物标志物的相关性。本研究的目的是确定苯并[a]芘(B[a]P)诱导的 DNA 加合物在斑马贻贝中的稳定性。贻贝暴露于溶解的 B[a]P(10-100μg/L)中 4 天。之后,贻贝在清洁水中饲养 28 天,随后使用(32)P-后标记技术在两个不同的器官(消化腺和鳃)中测量 DNA 加合物水平。同时,通过 qPCR(过氧化氢酶、超氧化物歧化酶、谷胱甘肽 S 转移酶、HSP70、芳烃受体、P 糖蛋白)评估解毒系统相关基因的表达。我们观察到消化腺中的 DNA 加合物水平高于鳃。此外,在鳃中,DNA 加合物的水平取决于 B[a]P 浓度。在清洁水中饲养 28 天后,两种器官中的加合物水平均呈下降趋势。此外,与消化腺相比,在鳃中观察到 HSP70、PgP、AHR 和 SOD mRNA 水平的早期诱导。CAT 和 GST 基因在两种器官中的表达均在 12 小时暴露时增加。除了 AHR 和 CAT 基因外,这些基因在鳃中的表达水平更高。数据表明 DNA 加合物是 B[a]P 污染和多环芳烃暴露的非常有前途的生物标志物。此外,在这项研究中,首次在 D. polymorpha 中表征了 B[a]P 解毒系统。