Polycyclic aromatic hydrocarbon induction of cytochrome P-450-dependent mixed-function oxidases in marine fish

doi:10.1016/0041-008X(80)90012-5

Toxicology and Applied Pharmacology

Volume 54, Issue 1, 15 June 1980, Pages 117-133

https://doi.org/10.1016/0041-008X(80)90012-5 Get rights and content

Abstract

Administration of 3-methylcholanthrene (3-MC) and related chemicals (1,2,3,4-dibenzanthracene, 7,12-dimethylbenzanthracene, and 5,6-benzoflavone) by ip or im injection to five species of marine fish resulted in increases in their hepatic microsomal mixed-function oxidase activities with benzo(a)pyrene, 7-ethoxycoumarin, and 7-ethoxyresorufin as substrates. Cytochrome P-450 content was increased in some treated fish. Sheepshead injected im with 3-MC showed the most marked induction of cytochrome P-450 content. In sheepshead, the Soret maximum of the CO-reduced difference spectrum changed from 450 nm in control to 448 or 449 nm in 3-MC-treated fish. The time course of 3-MC induction of benzo(a)pyrene hydroxylase activity in sheepshead changed with season (water temperature) and was not the same as the time course of induction of cytochrome P-450 content or 7-ethoxyresorufin O-deethylase activity. In summer (water temperature 26°C), benzo(a)pyrene hydroxylase activities in hepatic microsomes from 3-MC-treated sheepshead were maximum at 3 days and were back to control levels at 14 days after the dose, whereas in winter (water temperature 14°C) maximum activities were observed 8 days after dosing, but elevated activities were still observed 28 days after one ip dose. Epoxide hydrase and glutathione S-transferase activities were not induced by polycyclic aromatic hydrocarbons or 5,6-benzoflavone in the marine fish studied, at the doses tested.

References (44)

R.F. Addison et al.
Mixed-function oxidase enzymes in trout (Salvelinus fontinalis) liver; absence of induction following feeding of p,p′-DDT or p,p′-DDE
Comp. Biochem. Physiol.
(1977)
E. Bresnick et al.
Effect of phenobarbital and 3-methylcholanthrene administration on epoxide hydrase levels in liver microsomes
Biochem. Pharmacol.
(1977)
G. Clifton et al.
Effect of dietary phenobarbital, 3,4-benzo(a)pyrene and 3-methylcholanthrene on hepatic, intestinal and renal glutathione S-transferase activities in the rat
Biochem. Pharmacol.
(1978)
J.W. DePierre et al.
A reliable, sensitive and convenient radioactive assay for benzpyrene monoxygenase
Anal. Biochem.
(1975)
C.R. Elcombe et al.
Induction and characterization of hemoprotein(s) P-450 and monoxygenation in rainbow trout (Salmo gairdneri)
Toxicol. Appl. Pharmacol.
(1979)
E.H. Gerhart et al.
Hepatic mixed-function oxidase activity in rainbow trout exposed to several polycyclic aromatic compounds
Environ. Res.
(1978)
R.M. Harrison et al.
Polynuclear aromatic hydrocarbon in raw, potable, and waste waters
Water Res.
(1975)
W.H. Habig et al.
Glutathione S-transferases. The first enzymatic step in mercapturic acid formation
J. Biol. Chem.
(1974)
D.A. Haugen et al.
Induction of multiple forms of mouse liver cytochrome P-450
J. Biol. Chem.
(1976)
M.O. James et al.
Hepatic and extrahepatic metabolism of an epoxide (styrene oxide) by the rabbit
Biochem. Pharmacol.
(1976)

M.O. James et al.

Hepatic microsomal mixed-function oxidase activities in several marine species common to coastal Florida

Comp. Biochem. Physiol.

(1979)

M.O. James et al.

Epoxide hydrase and glutathione S-transferase activities with selected alkene and arene oxides in several marine species

Chem. Biol. Interact.

(1979)

E.F. Johnson et al.

Multiple forms of cytochrome P-450: Resolution and purification of rabbit liver aryl hydrocarbon hydroxylase

Biochem. Biophys. Res. Commun.

(1977)

H. Mukhtar et al.

Effect of phenobarbital and 3-methylcholanthrene administration on glutathione-S-epoxide transferase activity in rat liver

Biochem. Pharmacol.

(1976)

S. Nesnow et al.

An improved radiochemical assay for benzo(a)pyrene monooxygenase

Anal. Biochem.

(1977)

F. Oesch et al.

A radiometric assay for hepatic epoxide hydrase activity with (7-³H) styrene oxide

Biochim. Biophys. Acta

(1971)

T. Omura et al.

The carbon monoxide binding pigment of liver microsomes

J. Biol. Chem.

(1964)

D. Ryan et al.

Multiple forms of cytochrome P-450 in phenobarbital and 3-methylcholanthrene-treated rats

J. Biol. Chem.

(1975)

L. Balk et al.

Initial characterization of drug-metabolizing systems in the liver of the northern pike, Esox lucius

Drug. Metab. Disp.

(1980)

J.R. Bend et al.

Xenobiotic metabolism in marine and freshwater species

J.R. Bend et al.

Further studies of the microsomal mixed-function oxidase system of the little skate, Raja erinacea, including its response to some xenobiotics

Bull. Mt. Desert Island Biol. Lab.

(1973)

J.R. Bend et al.

Hepatic and extrahepatic glutathione S-transferase activity towards several arene oxides and epoxides in the rat

Cited by (117)

Consequences of prenatal exposure to contaminants in elasmobranchs: Biochemical outcomes during the embryonic development of Pseudobatos horkelii
2023, Environmental Pollution
Coastal elasmobranchs are vulnerable to chemicals mostly due to their k-strategic life history characteristics and high trophic positions. Embryos might be particularly exposed through the maternal offloading of contaminants, possibly leading to disruptions during critical developmental phases. Yet, knowledge on biochemical outcomes of prenatal exposure in elasmobranchs is notably limited. Therefore, we aimed to investigate the effects of prenatal exposure to contaminants in embryos of the critically endangered Brazilian guitarfish, Pseudobatos horkelii. Polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), pharmaceuticals and personal care products, and metals were determined in embryos. Additionally, glutathione S-transferase activity (GST), glutathione (GSH), and metallothionein levels (MT) were analyzed. Finally, lipid peroxidation levels (LPO) and protein carbonyl groups (PCO) were assessed. Embryonic exposure depended on yolk consumption, which was conspicuous in earlier development. We observed a dilution effect of contaminants levels, potentially related to biotransformation of these compounds throughout the embryonic development. Nevertheless, GST was not correlated to contaminant concentrations. The multivariate relationship between antioxidant components (GSH and GST) and LPO and PCO was negative, suggesting the lack of efficient defense of these biomarkers in early development, leading to oxidative damage. In this context, our results indicate that prenatal exposure to contaminants might impact the redox status in embryos of P. horkelii, leading to oxidative damage. Furthermore, metal concentrations influenced MT levels, suggesting this as a potential detoxification pathway in this species.
A multi-year study of hepatic biomarkers in coastal fishes from the Gulf of Mexico after the Deepwater Horizon Oil Spill
2017, Marine Environmental Research
Citation Excerpt :
These results suggest persistent exposure to petroleum compounds occurred for some reef fishes long after the well was capped in July 2010. Laboratory studies have shown that fish exposed to a single inducing dose of an AhR agonist exhibit elevated hepatic EROD activity for weeks after the dose, before a return to baseline levels (James and Bend, 1980; Kloepper-Sams and Stegeman, 1989). Field studies have validated the use of EROD as a biomarker of PAH exposure, both from oil spills and natural seepage (Moore et al., 2003; Morales-Caselles et al., 2006).
Following the 2010 Gulf of Mexico oil spill, concerns were raised regarding exposure of fish to crude oil components, particularly polycyclic aromatic hydrocarbons (PAHs). This three year study examined hepatic enzymes in post-mitochondrial supernatant fractions from red snapper (Lutjanus campechanus) and gray triggerfish (Balistes capriscus) collected in the north central Gulf of Mexico between 2011 and 2014. Biomarker activities evaluated included benzo(a)pyrene hydroxylase (AHH), ethoxyresorufin O-deethylase (EROD), glutathione transferase (GST), and glutathione peroxidase (GPx). Mean EROD activity was higher in gray triggerfish (12.97 ± 7.15 pmol/min/mg protein [mean ± SD], n = 115) than red snapper (2.75 ± 1.92 pmol/min/mg protein, n = 194), p < 0.0001. In both species, EROD declined over time between 2011 and 2014. Declines in GST and GPx activities were also noted over this time period for both species. Gray triggerfish liver was fatty, and heptane extracts of the liver fat contained fluorescent substances with properties similar to known PAHs, however the origin of these PAHs is unknown.
Artificial neural network modeling of biomarkers to infer characteristics of contaminant exposure in Clarias gariepinus
2012, Ecotoxicology and Environmental Safety
Citation Excerpt :
This study also demonstrated the power of ANN in predicting the route of exposure in fish biomarker studies, since small differences were observed between the computed and measured injection methods (Table 4). Route of exposure strongly influences fish biomarkers (James and Bend, 1980; Jonsson et al., 2004; Karami et al., 2011). Though natural routes of contaminant exposure (i.e., food, water, sediment) were not examined in the present study, the ability of ANN to differentiate different exposure routes in the lab (i.m. vs. i.p. injection) is encouraging.
This study examined the potential of artificial neural network (ANN) modeling to infer timing, route and dose of contaminant exposure from biomarkers in a freshwater fish. Hepatic glutathione S-transferase (GST) activity and biliary concentrations of BaP, 1-OH BaP, 3-OH BaP and 7,8D BaP were quantified in juvenile Clarias gariepinus injected intramuscularly or intraperitoneally with 10–50 mg/kg benzo[a]pyrene (BaP) 1–3 d earlier. A feedforward multilayer perceptron (MLP) ANN resulted in more accurate prediction of timing, route and exposure dose than a linear neural network or a radial basis function (RBF) ANN. MLP sensitivity analyses revealed contribution of all five biomarkers to predicting route of exposure but no contribution of hepatic GST activity or one of the two hydroxylated BaP metabolites to predicting time of exposure and dose of exposure. We conclude that information content of biomarkers collected from fish can be extended by judicious use of ANNs.
The effects of intramuscular and intraperitoneal injections of benzo[a]pyrene on selected biomarkers in Clarias gariepinus
2011, Ecotoxicology and Environmental Safety
Citation Excerpt :
We did not measure DNA adduct formation in the present study but the strong relationship between 7,8D BaP formation and dose and method of injection relative to other biliary FACs (partial eta squared=0.856 and 0.626, respectively; Table 1) demonstrates its potential as a biomarker in ecotoxicological studies. No significant difference between the GST activities in fish injected i.p. and i.m. is inconsistent with the study of James and Bend (1980) in which i.m. injection of 3-methylcholanthrene (3-MC) caused a higher CYP450 content than i.p. injection in five marine fish species. This difference may reflect the different PAHs used in the two studies but we did observe a different trend in GST activity between i.p.-and i.m.-injected fish.
This study investigated the dose-dependent and time-course effects of intramuscular (i.m.) and intraperitoneal (i.p.) injection of benzo[a]pyrene (BaP) on the biomarkers EROD activity, GST activity, concentrations of BaP metabolites in bile, and visceral fat deposits (Lipid Somatic Index, LSI) in African catfish (Clarias gariepinus). Intraperitoneal injection resulted in 4.5 times higher accumulation of total selected biliary FACs than i.m. injection. Hepatic GST activities were inhibited by BaP via both injection methods. Dose–response relationships between BaP injection and both biliary FAC concentrations and hepatic GST activities were linear in the i.p. injected group but nonlinear in the i.m. injected fish. Hepatic EROD activity and LSI were not significantly affected by BaP exposure by either injection route. We conclude that i.p. is a more effective route of exposure than i.m. for future ecotoxicological studies of PAH exposure in C. gariepinus.
Activity of Phase I and Phase II enzymes of the benzo[a]pyrene transformation pathway in zebrafish (Danio rerio) following waterborne exposure to arsenite
2010, Comparative Biochemistry and Physiology - C Toxicology and Pharmacology
The environmental pollutants inorganic arsenic (iAs) and benzo[a]pyrene (B[a]P) are carcinogens often found together in groundwater. The hepatic metabolism of B[a]P is a multi-step process requiring several Phase I and Phase II enzymes, notably cytochrome p450 1A (CYP1A), epoxide hydrolase (EH), and glutathione S-transferase (GST). The purpose of this study was to examine the effect of arsenite (As(III)) on the activity of these enzymes in vivo utilizing adult zebrafish (Danio rerio). Zebrafish were exposed to either 0.4 μM B[a]P, 0.4 μM B[a]P + 0.4 μM As(III), 0.4 μM B[a]P + 8 μM As(III), 0.4 μM As(III), or 8 μM As(III) for 7 days. Co-exposures to As(III) and B[a]P led to significant decreases in CYP1A enzyme activity (approximately 3-fold) when compared to exposure to B[a]P alone. No similar effects occurred with EH or GST, although B[a]P exposure did significantly increase EH activity. Furthermore As(III) and B[a]P co-exposures significantly decreased CYP1A transcript levels (up to 35-fold) when compared to B[a]P. However, B[a]P-induced CYP1A protein levels remained elevated following co-exposures to As(III). This evidence suggests that As(III) has the potential to modify components of the B[a]P biotransformation pathway in vivo via a disruption of CYP1A activity by way of both pre- and post-translational mechanisms.
Impaired reproductive health of killifish (Fundulus heteroclitus) inhabiting Newark Bay, NJ, a chronically contaminated estuary
2010, Aquatic Toxicology
A battery of biomarkers were used to evaluate the reproductive health and contaminant exposure of Atlantic killifish (Fundulus heteroclitus) inhabiting the heavily industrialized Newark Bay and a reference population from Great Bay, Tuckerton, NJ. The biomarkers investigated included classical endpoints (gonad and liver histopathology, body and tissue morphometrics), hepatic mRNA expression (CYP1A and vitellogenin I), hepatic protein levels (CYP1A and vitellogenin), gonadal aromatase mRNA expression, and chemical exposure analyses (bile PAHs). Our data showed no significant differences between populations for body size and body weight. However, Newark Bay killifish exhibited molecular and morphological changes indicative of impaired reproductive health and endocrine disruption compared to the reference population. Newark Bay males had decreased gonad weight, altered testis development and decreased gonadal aromatase mRNA expression. Newark Bay females had decreased gonad weight, inhibited gonadal development, decreased hepatic vitellogenin production (mRNA and protein) and increased mRNA expression of gonadal aromatase. In addition, Newark Bay females had a significant increase in the percent of pre-vitellogenic follicles (43% at Tuckerton, 64% at Newark Bay) and a significantly decreased percent of follicles at the mid-vitellogenic and mature stages (25% mature at Tuckerton and 3% at Newark Bay). In addition to reproductive endpoints, killifish at Newark Bay exhibited high basal levels of CYP1A mRNA and protein expression which indicated exposure to aryl hydrocarbon receptor (AhR) agonists. An inverse relationship between hepatic CYP1A protein and hepatic vitellogenin mRNA expression was established suggesting a possible link between AhR agonist exposure and vitellogenesis. Killifish in the NY–NJ Harbor Estuary are exposed to a number of chemicals that can interact with the AhR pathway and stimulate enzymatic activity along with chemicals that can modify reproductive success in this indigenous species. Similar effects on the reproductive development in less resilient species may limit their ability to repopulate the NY–NJ Harbor Estuary and similarly contaminated water systems.

View all citing articles on Scopus

View full text

Polycyclic aromatic hydrocarbon induction of cytochrome P-450-dependent mixed-function oxidases in marine fish

Abstract

Comp. Biochem. Physiol.

Biochem. Pharmacol.

Biochem. Pharmacol.

Anal. Biochem.

Toxicol. Appl. Pharmacol.

Environ. Res.

Water Res.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Pharmacol.

Comp. Biochem. Physiol.

Chem. Biol. Interact.

Biochem. Biophys. Res. Commun.

Biochem. Pharmacol.

Anal. Biochem.

Biochim. Biophys. Acta

J. Biol. Chem.

J. Biol. Chem.

Initial characterization of drug-metabolizing systems in the liver of the northern pike, Esox lucius

Drug. Metab. Disp.

Xenobiotic metabolism in marine and freshwater species

Further studies of the microsomal mixed-function oxidase system of the little skate, Raja erinacea, including its response to some xenobiotics

Bull. Mt. Desert Island Biol. Lab.

Hepatic and extrahepatic glutathione S-transferase activity towards several arene oxides and epoxides in the rat