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.
| Original language | English |
|---|---|
| Pages (from-to) | 282-289 |
| Number of pages | 8 |
| Journal | Chemosphere |
| Volume | 107 |
| DOIs | |
| Publication status | Published - 2014 |
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Keywords
- Antibodies
- Benzo[a]pyrene
- Colocalization
- Dioxin
- Earthworms
- Immunofluorescence
ASJC Scopus subject areas
- Environmental Chemistry
- Chemistry(all)
Cite this
Immunofluorescence detection and localization of B[a]P and TCDD in earthworm tissues. / Sforzini, Susanna; Moore, Michael N.; Boeri, Marta; Benfenati, Emilio; Colombo, Andrea; Viarengo, Aldo.
In: Chemosphere, Vol. 107, 2014, p. 282-289.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Immunofluorescence detection and localization of B[a]P and TCDD in earthworm tissues
AU - Sforzini, Susanna
AU - Moore, Michael N.
AU - Boeri, Marta
AU - Benfenati, Emilio
AU - Colombo, Andrea
AU - Viarengo, Aldo
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - Antibodies
KW - Benzo[a]pyrene
KW - Colocalization
KW - Dioxin
KW - Earthworms
KW - Immunofluorescence
UR - http://www.scopus.com/inward/record.url?scp=84901589154&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84901589154&partnerID=8YFLogxK
U2 - 10.1016/j.chemosphere.2013.12.062
DO - 10.1016/j.chemosphere.2013.12.062
M3 - Article
C2 - 24412505
AN - SCOPUS:84901589154
VL - 107
SP - 282
EP - 289
JO - Chemosphere
JF - Chemosphere
SN - 0045-6535
ER -