The formation of DNA adducts was evident after treatment of spawned or resting Mediterranean mussels (Mytilus galloprovincialis Lmk.) with 0.5-100 mu g/l of benzo[a]pyrene for 2 and 3 days. Reference DNA samples, in vitro radiolabelled with 0.5, 5, 50 mu M H-3-anti (+/-)-B[a]P-diol-epoxide, were initially used to compare two DNA purification procedures. Following a standard four-step extraction starting with phenol, in comparison to a simplified single-step extraction without phenol, we obtained lower yields of bound radioactivity in the reference DNA samples. After simplified DNA purification and nuclease P1 enhanced P-32-postlabelling assay we detected a reproducible dose-dependent increase of a specific spot in gills of mussels treated with B[a]P, although this spot was present in low amounts. Short (2 days) and prolonged (27 days) pretreatment of mussels with a polychlorinated biphenyl mixture, Aroclor 1254, did not increase the levels of B[a]P-related adducts. On the whole, these results indicate the formation of detectable amounts of DNA reactive intermediates in gills of mussels treated with B[a]P. Although the pathway of formation and the molecular identity of the specific adducts remain unclear, their presence suggests that polycyclic aromatic hydrocarbons may cause genetic damage in marine mussels.

Formation of DNA adducts in the gill tissue of Mytilus galloprovincialis treated with benzo[a]pyrene.

VENIER, PAOLA;CANOVA, SABRINA
1996

Abstract

The formation of DNA adducts was evident after treatment of spawned or resting Mediterranean mussels (Mytilus galloprovincialis Lmk.) with 0.5-100 mu g/l of benzo[a]pyrene for 2 and 3 days. Reference DNA samples, in vitro radiolabelled with 0.5, 5, 50 mu M H-3-anti (+/-)-B[a]P-diol-epoxide, were initially used to compare two DNA purification procedures. Following a standard four-step extraction starting with phenol, in comparison to a simplified single-step extraction without phenol, we obtained lower yields of bound radioactivity in the reference DNA samples. After simplified DNA purification and nuclease P1 enhanced P-32-postlabelling assay we detected a reproducible dose-dependent increase of a specific spot in gills of mussels treated with B[a]P, although this spot was present in low amounts. Short (2 days) and prolonged (27 days) pretreatment of mussels with a polychlorinated biphenyl mixture, Aroclor 1254, did not increase the levels of B[a]P-related adducts. On the whole, these results indicate the formation of detectable amounts of DNA reactive intermediates in gills of mussels treated with B[a]P. Although the pathway of formation and the molecular identity of the specific adducts remain unclear, their presence suggests that polycyclic aromatic hydrocarbons may cause genetic damage in marine mussels.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11577/124366
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