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ToxSci Advance Access published online on February 14, 2008
Toxicological Sciences, doi:10.1093/toxsci/kfn024
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© The Author 2008. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

Surface marker-defined head kidney granulocytes and B-lymphocytes of rainbow trout express benzo[a]pyrene-inducible cytochrome P4501A protein

Ayako Nakayama*,§, Ivan Riesen*,§, Bernd Köllner{dagger}, Elisabeth Eppler{ddagger} and Helmut Segner*

* Centre for Fish and Wildlife Health, University of Berne, P.O. Box 8466, CH-3001 Berne (Switzerland), ayako.nakayama{at}itpa.unibe.ch, ivan.riesen{at}itpa.unibe.ch {dagger} Friedrich-Loeffler-Institute, Federal Research Institute for Animal Health, D-17493 Greifswald-Insel Riems (Germany), bernd.koellner{at}fli.bund.de {ddagger} Research Group Neuro-Endocrine-Immune Interactions, Division of Neuroendocrinology, Institute of Anatomy, University of Zürich, Winterthurerstr. 190, CH-8057 Zürich (Switzerland), eppler{at}anatom.uzh.ch

Corresponding author: Helmut Segner, Email address: helmut.segner{at}itpa.unibe.ch, Centre for Fish and Wildlife Health, University of Berne, P.O. Box 8466, CH-3001 Berne (Switzerland), Telephone No.: +41 (0)31-631-2441, Fax No.: +41-(0)31-631-2611

Received October 26, 2007; revision received January 31, 2008; accepted February 1, 2008


  Abstract

Polycyclic aromatic hydrocarbons (PAHs) such as benzo[a]pyrene (BaP) are immunotoxic to fish. Metabolism of PAHs in immune cells has been implicated in PAH immunotoxicity in mammals, but for fish the presence of metabolic enzymes in immune cells is less clear. The objective of this study was to examine localisation and induction of the BaP-metabolizing biotransformation enzyme, cytochrome P4501A, in head kidney immune cells of rainbow trout (Oncorhynchus mykiss). In the first step, we measured induction of CYP1A-dependent 7-ethoxyresorufin-O-deethylase (EROD) activity and CYP1A protein in head kidney of rainbow trout treated with a single intraperitoneal injection of 25 mg BaP/kg body weight. From days 3 to 10 post-injection, the BaP treatment led to a significant elevation of EROD and CYP1A protein in head kidney and liver, with CYP1A expression levels in the head kidney being much lower than in the liver. Next, we examined the cellular localisation of CYP1A protein in the head kidney cell types: vascular endothelial, endocrine and lymphoid cells. CYP1A immunoreactivity was detectable only in BaP-treated trout, where it was localised in endothelial and lymphoid cells. Finally, we aimed to clarify which of the hematopoietic cell types possess CYP1A protein. Using double immunostaining for CYP1A and surface markers of rainbow trout immune cells, we identified B-lymphocytes and granulocytes expressing inducible CYP1A protein and being the likely sites of BaP metabolism in the head kidney

Key Words: rainbow trout; benzo[a]pyrene; immunotoxicity; cytochrome P4501A; granulocytes; B-lymphocytes.

§ These authors contributed equally to the work


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