Oxidative damage to DNA: an immunohistochemical approach for detection of 7,8-dihydro-8-oxodeoxyguanosine in marine organisms.
The modified nucleoside 7,8-dihydro-8-oxodeoxyguanosine (8-oxo-dG) is an index of oxidative DNA damage. An immunohistochemical approach based on the use of monoclonal antibody 1F7 against 8-oxo-dG was investigated in marine organisms with immunoperoxidase and immunofluorescent detection. Relative staining intensity as a measure of the 8-oxo-dG level was microscopically assessed. After laboratory exposures to benzo[a]pyrene (B[a]P), higher levels of oxidative DNA damage were clearly detected in all treated animals compared to controls. While this method eliminates DNA extraction reducing the processing of biological samples, absolute values are not provided. Further, the method requires only small amounts of tissue and potentially discriminates susceptibility to oxidative damage in different cell types. These results suggest that the assay should have practical applications in marine ecotoxicology.
Machella
Nicola
N
Istituto di Biologia e Genetica, Università Politecnica delle Marche, Via Ranieri 65, Monte d'Ago, 60100 Ancona, Italy. nicola_machella@yahoo.com
Regoli
Francesco
F
Cambria
Antonio
A
Santella
Regina M
RM
D000811:Anguilla / Q000235:genetics* / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001564:Benzo(a)pyrene / Q000633:toxicity*; D049872:Bivalvia / Q000235:genetics* / Q000276:immunology; D004249:DNA Damage / Q000187:drug effects* / Q000235:genetics; D005060:Europe; D005455:Fluorescent Antibody Technique; D006151:Guanosine / Q000031:analogs & derivatives*; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods
D000911:Antibodies, Monoclonal; D006151:Guanosine; D001564:Benzo(a)pyrene; C046215:8-hydroxyguanosine