Monoclonal antibodies to liver microsomal cytochrome P-450E of the marine fish Stenotomus chrysops (scup): Cross reactivity with 3-methylcholanthrene induced rat cytochrome P-450

Park, Sang S.; Miller, Haruko; Klotz, Alan V.; Kloepper-Sams, Pamela J.; Stegeman, John J.; Gelboin, Harry V.

doi:10.1016/0003-9861(86)90010-x

Cited by 180 publications

(64 citation statements)

References 27 publications

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“…Equivalent volumes of lysate were subjected to SDS-PAGE and blotted to nitrocellulose. Blots were probed with monoclonal antibody 1-12-3 (3 g/mL) against Stenotomus chrysops (scup) CYP1A (Park et al, 1986;Kloepper-Sams et al, 1987). Blots were subsequently probed with goat anti-mouse IgG horseradish peroxidase (Bio-Rad, Hercules, CA) secondary antibody (1:1000 dilution).…”

Section: Western Blot Analysismentioning

confidence: 99%

Development of the morpholino gene knockdown technique in Fundulus heteroclitus: A tool for studying molecular mechanisms in an established environmental model

et al. 2008

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a b s t r a c t A significant challenge in environmental toxicology is that many genetic and genomic tools available in laboratory models are not developed for commonly used environmental models. The Atlantic killifish (Fundulus heteroclitus) is one of the most studied teleost environmental models, yet few genetic or genomic tools have been developed for use in this species. The advancement of genetic and evolutionary toxicology will require that many of the tools developed in laboratory models be transferred into species more applicable to environmental toxicology. Antisense morpholino oligonucleotide (MO) gene knockdown technology has been widely utilized to study development in zebrafish and has been proven to be a powerful tool in toxicological investigations through direct manipulation of molecular pathways. To expand the utility of killifish as an environmental model, MO gene knockdown technology was adapted for use in Fundulus. Morpholino microinjection methods were altered to overcome the significant differences between these two species. Morpholino efficacy and functional duration were evaluated with molecular and phenotypic methods. A cytochrome P450-1A (CYP1A) MO was used to confirm effectiveness of the methodology. For CYP1A MO-injected embryos, a 70% reduction in CYP1A activity, a 86% reduction in total CYP1A protein, a significant increase in ␤-naphthoflavone-induced teratogenicity, and estimates of functional duration (50% reduction in activity 10 dpf, and 86% reduction in total protein 12 dpf) conclusively demonstrated that MO technologies can be used effectively in killifish and will likely be just as informative as they have been in zebrafish.

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Section: Western Blot Analysismentioning

confidence: 99%

Development of the morpholino gene knockdown technique in Fundulus heteroclitus: A tool for studying molecular mechanisms in an established environmental model

et al. 2008

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“…The sections were inserted into Shandon coverplates (Shandon Lipshaw Inc., Pittsburg, PA) and incubated with normal goat serum for 5 min to block nonspecific binding of the secondary antibody. This was followed by two 1-h incubations with 150 ml of monoclonal antibody 1-12-3 (Park et al, 1986). The specificity of this antibody for CYP1A has been shown previously (Miller et al, 1989).…”

Section: Cytochrome P450 Assaymentioning

confidence: 99%

2,3,7,8-Tetrachlorodibenzo-p-dioxin induces apoptotic cell death and cytochrome P4501A expression in developing Fundulus heteroclitus embryos

et al. 2001

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“…Some embryos stained by in situ hybridization were sectioned and counter-stained with eosin for detailed observation, after being embedded in paraffin. Conventional immunohistochemistry for CYP1A was performed using a monoclonal antibody specific for fish CYP1A (× 2,400, Mab 1-12-3) [11], according to Teraoka et al [16]. The sections were counter-stained with methyl green after coloring reactions with diaminobenzidine.…”

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confidence: 99%

cDNA Cloning and Expressions of Cytochrome P450 1A in Zebrafish Embryos.

Yamazaki

Teraoka

Wang

et al. 2002

The Journal of Veterinary Medical Science

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ABSTRACT. Cytochrome P450 1A (CYP1A) is well known for being induced by aromatic hydrocarbons, including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). We determined the complete cDNA sequence of a CYP1A open reading frame with both 5'-and 3'-ends in zebrafish (zfCYP1A), a useful model for environmental toxicology. zfCYP1A shows high percentage identity with CYP1As of mammals, domestic fowl and xenopus (51.9-60.4%), as well as the other fish species (63.8-89.2%). As revealed by in situ hybridization and immunohistochemistry, zfCYP1A was scarcely detected in control embryos but was markedly induced by TCDD especially in heart, vascular endothelial cells, intestinal epithelium, pronephros and outer integument in both prehatched and hatched embryos. These e xpression patterns are consistent with possible involvement of zfCYP1A in TCDD-induced toxicities. KEY WORDS: CYP1A, TCDD, zebrafish.

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Monoclonal antibodies to liver microsomal cytochrome P-450E of the marine fish Stenotomus chrysops (scup): Cross reactivity with 3-methylcholanthrene induced rat cytochrome P-450

Cited by 180 publications

References 27 publications

Development of the morpholino gene knockdown technique in Fundulus heteroclitus: A tool for studying molecular mechanisms in an established environmental model

Development of the morpholino gene knockdown technique in Fundulus heteroclitus: A tool for studying molecular mechanisms in an established environmental model

2,3,7,8-Tetrachlorodibenzo-p-dioxin induces apoptotic cell death and cytochrome P4501A expression in developing Fundulus heteroclitus embryos

cDNA Cloning and Expressions of Cytochrome P450 1A in Zebrafish Embryos.

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