Expression of HSP70 and CYP1A protein in ovary and liver of juvenile rainbow trout exposed to beta-naphthoflavone.
Cytochrome P4501A (CYP1A) and the 70-kDa stress protein (HSP70) were determined using Western blotting in the ovary and liver of juvenile female rainbow trout (Oncorhynchus mykiss) exposed for 4 days to beta-naphthoflavone (betaNF) following a single intraperitoneal injection. Ovarian CYP1A protein was observed in both control and betaNF-exposed fish, indicating constitutive and inducible expression of CYP1A in immature trout ovaries. CYP1A protein levels determined using densitometry were 14- and 46-fold greater in betaNF-exposed trout compared to controls in the liver and ovary, respectively. Hepatic microsomal ethoxyresorufin O-deethylase (EROD) activity, a specific catalytic marker of CYP1A, was also induced 38-fold above controls following betaNF exposure. Hepatic HSP70 protein expression was significantly higher in whole cell homogenates, but not in cytosolic fractions, collected from betaNF-exposed fish in comparison to control fish. There was no difference in ovarian HSP70 levels determined in whole cell homogenates between control and betaNF-exposed fish. The observation that unlike liver, ovarian HSP70 expression remained unchanged following induction of CYP1A protein may be related to the sensitivity of the teleost ovary to environmental toxicants that act as aryl hydrocarbon receptor agonists.
Weber
Lynn P
LP
Department of Zoology, Oklahoma State University, 430 Life Sciences West, Stillwater, OK 74078, USA.
Diamond
Stephen L
SL
Bandiera
Stelvio M
SM
Janz
David M
DM
D000818:Animals; D000918:Antibody Specificity; D019363:Cytochrome P-450 CYP1A1 / Q000032:analysis / Q000096:biosynthesis; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis / Q000096:biosynthesis*; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000096:biosynthesis*; D007527:Isoenzymes / Q000032:analysis / Q000096:biosynthesis; D008099:Liver / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D008862:Microsomes, Liver / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D017686:Oncorhynchus mykiss; D010053:Ovary / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D020033:Protein Isoforms / Q000032:analysis / Q000096:biosynthesis; D019324:beta-Naphthoflavone / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D018840:HSP70 Heat-Shock Proteins; D007527:Isoenzymes; D020033:Protein Isoforms; D019324:beta-Naphthoflavone; D003577:Cytochrome P-450 Enzyme System; D019363:Cytochrome P-450 CYP1A1