Expression of P4501A1 in a primary culture of rainbow trout hepatocytes exposed to β-naphthoflavone or 2,3,7,8-tetrachlorodibenzo-p-dioxin

Pesonen, Maija; Goksøyr, Anders; Andersson, Tommy

doi:10.1016/0003-9861(92)90072-5

Cited by 93 publications

(44 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The fact that the pattern of DNA adducts was the same as that observed in the absence of EROD induction (see experiment II) supports the assumption that P4501A was present in control hepatocytes, as recently shown by Pesonen et al (1992) and that EROD induction did not result from the synthesis of new forms of P-450 involved in B[a]P activation, but from reinforcement of the formation/stabilization of P-4501A present in the cells before treatment with flNF.…”

Section: Discussionsupporting

confidence: 86%

“…Unfortunately, these authors did not describe the effect of this concentration of B[a]P on the activity of P4501A. As, in our study, EROD was not induced after 12hr of exposure, it seems likely that activation of B[a]P by rainbow trout hepatocytes takes place before any significant increase in P4501A protein content, which occurs after some delay caused by the synthesis of CYP1A mRNA, as recently shown by Pesonen et al (1992). A close similarity was shown between DNA adduct patterns generated in cultured hepatocytes and in vivo, suggesting that the metabolic activation of PAHs in primary culture of rainbow trout hepatocytes is representative of that in vivo.…”

Section: Discussionsupporting

confidence: 60%

See 1 more Smart Citation

DNA adduct formation and 7-ethoxyresorufin O-deethylase induction in primary culture of rainbow trout hepatocytes exposed to benzo[a]pyrene

Masfaraud¹,

Devaux²,

Pfohl‐Leszkowicz³

et al. 1992

Toxicology in Vitro

View full text Add to dashboard Cite

Abstract--The formation of DNA adducts, using the 32p-postlabeiling assay, and induction of 7-ethoxyresorufin O-deethylase (EROD) were investigated in a primary culture of rainbow trout hepatocytes exposed to benzo[a]pyrene (B[a]P). Concentrations of 0.1 and 1/zM-B[a]P were shown to induce EROD whereas 10 #M was an inhibitory concentration. DNA adducts were detected for 12 hr to 72 hr after exposure to 1 #M-B[a]P whereas EROD activity was increased 36 hr after treatment. The pattern of adducts was shown to be identical to that obtained after B[a]P treatment of rainbow trout in vivo, as demonstrated by co-chromatography of the adducts. Pre-exposure of hepatocytes for 48 hr to fl-naphthoflavone (~NF) and subsequent 24-hr exposure to 1 ~uM-B[a]P did not lead to increased DNA adduct formation although flNF treatment led to a 3.4-fold induction of EROD activity at the time of B[a]P addition. This study suggests that primary culture of rainbow trout hepatocytes provides a suitable method for studying DNA adduct formation and its modulating factors/n vitro.

show abstract

Section: Discussionsupporting

confidence: 86%

Section: Discussionsupporting

confidence: 60%

DNA adduct formation and 7-ethoxyresorufin O-deethylase induction in primary culture of rainbow trout hepatocytes exposed to benzo[a]pyrene

Masfaraud¹,

Devaux²,

Pfohl‐Leszkowicz³

et al. 1992

Toxicology in Vitro

View full text Add to dashboard Cite

show abstract

“…PB-induced accumulation of CYP1A1 mRNA decreased after peaking, and by 48 h the CYP1A1 mRNA levels remained static until the duration of the experiment (76 h). PB induction of EROD activity lagged behind the elevated CYP1A1 mRNA levels by approximately 9 h. The kinetics of PB induction of EROD activity and CYP1A1 mRNA levels were similar to that of ␤-naphthoflavone (␤NF) in primary cultures of rainbow trout hepatocytes (51). Conversely, the kinetics of TCDD induction of CYP1A1 mRNA in primary cultures of rainbow trout hepatocytes has been shown to increase linearly up to 12 h after TCDD addition and then remain at a plateau for up to 96 h (51).…”

Section: Cells Treated With Varying Concentrations Of Phenobarbital Imentioning

confidence: 79%

Phenobarbital Induction of Gene Expression in a Primary Culture of Rainbow Trout Hepatocytes

Sadar

Ash

Sundqvist

et al. 1996

Journal of Biological Chemistry

View full text Add to dashboard Cite

In mammals, phenobarbital (PB) is an in vivo inducer of the cytochrome P4502B (CYP2B) family, whereas in teleosts PB induction of cytochrome P450 is unclear. We show that teleost cytochrome P4502K1 (CYP2K1) protein levels and 7-pentoxyresorufin-O-deethylase activity were not induced by exposure of primary cultures of rainbow trout hepatocytes to PB. Instead, cytochrome P4501A1 (CYP1A1) gene expression was strongly induced by PB, based upon observations of marked increases in CYP1A1 mRNA, CYP1A1 protein, and 7-ethoxyresorufin-O-deethylase activity. In accordance with these data we provide a temporal study employing antibodies for the aromatic hydrocarbon (Ah) receptor that showed an increase in Ah receptor in nuclear extracts prepared from cells exposed to PB. Employment of the electrophoretic mobility shift assay (EMSA) showed PB to cause activation or "transformation" of the Ah receptor in nuclear extracts. Studies employing actinomycin D and cycloheximide indicated that PB induction of CYP1A1 was regulated at both the transcriptional and post-transcriptional levels. Nuclear run-off experiments confirm that PB causes an increase in CYP1A1 transcription. Inhibition of protein synthesis led to the superinduction of CYP1A1 mRNA, suggesting the regulation of teleost CYP1A1 may involve a labile repressor protein. These findings suggest that PB induction of the CYP1A1 gene involves the Ah receptor and is via transcriptional activation.

show abstract

“…The P450 1A induction kinetics in fish primary hepatocyte cultures, as shown in Fig. 2, were similar to that in in vivo tests with fish after a single treatment (Pesonen et al, 1992). It is possible that primary cells maintain intact characteristics of the original tissue in in vivo and many metabolic systems, including biotransformation of complex environmental matrices.…”

Section: Resultsmentioning

confidence: 51%

EROD activities in a primary cell culture of grass carp (Ctenopharyngodon idellus) hepatocytes exposed to polychlorinated aromatic hydrocarbonas

Wan

et al. 2004

Ecotoxicology and Environmental Safety

View full text Add to dashboard Cite

Aryl hydrocarbon (Ah) receptor (Ah-agonist) effects of environmental samples containing polychlorinated aromatic hydrocarbons were evaluated using a 7-ethoxyresorufin-O-deethylase (EROD) assay of a primary hepatocyte culture from grass carp (Ctenopharyngodon idellus). The results were compared with those obtained from the assay using the rat hepatoma cell line H4IIE and chemical analysis using high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS). A dose-response relationship was observed between the EROD activities, either from primary hepatocyte culture assay or from H4IIE assay, and concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The results showed that the assay based on the H4IIE cell line (EC 50 =0.83 pg/mL) is more sensitive to TCDD than the assay based on primary hepatocyte culture (EC 50 =9.7 pg/mL). In tests of environmental samples, the results from the assay using primary hepatocyte culture were comparable to those from the assay using the H4IIE cell line and chemical analysis of concentrations of mixtures of polychlorinated dibenzo-p-dioxin and dibenzofuran (PCDD/PCDF). The lack of a change in the activities of glutathione-S-transferase (GST) and lactate dehydrogenase (LDH) in cell culture upon exposure to TCDD indirectly indicates that the compound is persistent to biodegradation in the cell culture system. r 2004 Elsevier Inc. All rights reserved.

show abstract

Expression of P4501A1 in a primary culture of rainbow trout hepatocytes exposed to β-naphthoflavone or 2,3,7,8-tetrachlorodibenzo-p-dioxin

Cited by 93 publications

References 24 publications

DNA adduct formation and 7-ethoxyresorufin O-deethylase induction in primary culture of rainbow trout hepatocytes exposed to benzo[a]pyrene

DNA adduct formation and 7-ethoxyresorufin O-deethylase induction in primary culture of rainbow trout hepatocytes exposed to benzo[a]pyrene

Phenobarbital Induction of Gene Expression in a Primary Culture of Rainbow Trout Hepatocytes

EROD activities in a primary cell culture of grass carp (Ctenopharyngodon idellus) hepatocytes exposed to polychlorinated aromatic hydrocarbonas

Contact Info

Product

Resources

About