title: Immunofluorescence detection and localization of B[a]P and TCDD in earthworm tissues.
abstract: An immunohistochemical method using antibodies against polycyclic aromatic hydrocarbons (PAHs) and dioxins was developed on frozen tissue sections of the earthworm Eisenia andrei exposed to environmentally relevant concentrations of benzo[a]pyrene (B[a]P) (0.1, 10, 50 ppm) and 2,3,7,8-tetrachloro-dibenzo-para-dioxin (TCDD) (0.01, 0.1, 2 ppb) in spiked standard soils. The concentrations of B[a]P and TCDD in E. andrei exposed to the same conditions were also measured using analytical chemical procedures. The results demonstrated that tissues of worms exposed to even minimal amount of B[a]P and TCDD reacted positively and specifically to anti-PAHs and -dioxins antibody. Immunofluorescence revealed a much more intense staining for the gut compared to the body wall; moreover, positively immunoreactive amoeboid coelomocytes were also observed, i.e. cells in which we have previously demonstrated the occurrence of genotoxic damage. The double immunolabelling with antibodies against B[a]P/TCDD and the lysosomal enzyme cathepsin D demonstrated the lysosomal accumulation of the organic xenobiotic compounds, in particular in the cells of the chloragogenous tissue as well as in coelomocytes, involved into detoxification and protection of animals against toxic chemicals. The method described is timesaving, not expensive and easily applicable.
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "A. Avogadro", V.le T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Susanna
authors.initials: S
authors.lastname: Sforzini
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "A. Avogadro", V.le T. Michel 11, 15121 Alessandria, Italy; European Centre for Environment & Human Health (ECEHH), University of Exeter Medical School, Truro TR1 3HD, UK; Plymouth Marine Laboratory, Plymouth PL1 3DH, UK.
authors.forename: Michael N
authors.initials: MN
authors.lastname: Moore
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "A. Avogadro", V.le T. Michel 11, 15121 Alessandria, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Boeri
authors.affiliation: Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Via Giuseppe La Masa 19, 20156 Milan, Italy.
authors.forename: Emilio
authors.initials: E
authors.lastname: Benfenati
authors.affiliation: Department of Environmental Health Sciences, IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Via Giuseppe La Masa 19, 20156 Milan, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Colombo
authors.affiliation: Department of Sciences and Technological Innovation (DiSIT), University of Piemonte Orientale "A. Avogadro", V.le T. Michel 11, 15121 Alessandria, Italy. Electronic address: viarengo@unipmn.it.
authors.forename: Aldo
authors.initials: A
authors.lastname: Viarengo
mesh_terms: D000818:Animals; D001564:Benzo(a)pyrene / Q000378:metabolism*; D001692:Biological Transport; D002402:Cathepsin D / Q000378:metabolism; D004249:DNA Damage; D005455:Fluorescent Antibody Technique / Q000379:methods*; D009835:Oligochaeta / Q000378:metabolism*; D000072317:Polychlorinated Dibenzodioxins / Q000378:metabolism*; D012987:Soil / Q000737:chemistry
chemical_list: D000072317:Polychlorinated Dibenzodioxins; D012987:Soil; D001564:Benzo(a)pyrene; D002402:Cathepsin D
keywords: Antibodies; Benzo[a]pyrene; Colocalization; Dioxin; Earthworms; Immunofluorescence