Hepatic cellular distribution of cytochrome P-450 IA1 in rainbow trout (Oncorhynchus mykiss): An immunohisto- and cytochemical study

Susan M. Lester, Swee J Teh, Swee J. Teh, John J. Stegeman, Michael R. Miller, David E. Hinton

Research output: Contribution to journalArticle

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Abstract

Monoclonal antibody 1-12-3 reactive against scup (Stenotomus chrysops) cytochrome P450 E (a teleost CYP IA1) has been used to immunohistochemically localize CYP IA1 within hepatocytes and presumably sinusoidal endothelial and biliary epithelial cells of scup and trout. The goal of the present study was to extend immunohistochemical studies to the ultrastructural level determining intracellular locations of CYP IA1 in fish liver. Juvenile trout (5-10 g) were given i.p. injections once (50 μg/g β-naphthoflavone in cod liver oil; 0.5-ml injectate volume). After 5 days, livers were fixed (0.25% glutaraldehyde) via vascular in situ perfusion, removed, cut in 100-μm slices, infiltrated, and embedded in LR White monomer. Ultrathin sections of exposed livers were incubated in monoclonal antibody 1-12-3, rabbit anti-mouse IgG, and protein G colloidal gold. Membranes of granular endoplasmic reticulum in perinuclear regions of hepatocytes were consistently labeled. In addition, hepatocyte plasma membrane, particularly microvilli at bile canaliculi, was labeled. Biliary epithelial cells were labeled on luminal plasma membrane surrounding biliary passageway. Plasma membrane facing sinusoid and immediately subjacent cytoplasm was labeled in endothelial cells. Presence of CYP IA1 in sinusoidal endothelium could contribute to detoxication and/or bioactivation of blood borne chemicals. Granular endoplasmic reticulum was not uniformly labeled in hepatocytes. Rather, distribution seemed sequestered within highly specific regions and not dispersed along all membrane surfaces. Localization within biliary epithelial cells could signify potential of this cell type to bioactivate polycyclic aromatic hydrocarbons and may explain the common finding of biliary as well as hepatocytic tumors of trout liver.

Original languageEnglish (US)
Pages (from-to)3700-3706
Number of pages7
JournalCancer Research
Volume53
Issue number16
StatePublished - Aug 15 1993

Fingerprint

Oncorhynchus mykiss
Cytochrome P-450 Enzyme System
Trout
Hepatocytes
Rough Endoplasmic Reticulum
Liver
Epithelial Cells
Cell Membrane
Cod Liver Oil
Monoclonal Antibodies
Bile Canaliculi
Gold Colloid
Membranes
Polycyclic Aromatic Hydrocarbons
Glutaral
Microvilli
Endothelium
Blood Vessels
Cytoplasm
Fishes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Lester, S. M., Teh, S. J., Teh, S. J., Stegeman, J. J., Miller, M. R., & Hinton, D. E. (1993). Hepatic cellular distribution of cytochrome P-450 IA1 in rainbow trout (Oncorhynchus mykiss): An immunohisto- and cytochemical study. Cancer Research, 53(16), 3700-3706.

Hepatic cellular distribution of cytochrome P-450 IA1 in rainbow trout (Oncorhynchus mykiss) : An immunohisto- and cytochemical study. / Lester, Susan M.; Teh, Swee J; Teh, Swee J.; Stegeman, John J.; Miller, Michael R.; Hinton, David E.

In: Cancer Research, Vol. 53, No. 16, 15.08.1993, p. 3700-3706.

Research output: Contribution to journalArticle

Lester, Susan M. ; Teh, Swee J ; Teh, Swee J. ; Stegeman, John J. ; Miller, Michael R. ; Hinton, David E. / Hepatic cellular distribution of cytochrome P-450 IA1 in rainbow trout (Oncorhynchus mykiss) : An immunohisto- and cytochemical study. In: Cancer Research. 1993 ; Vol. 53, No. 16. pp. 3700-3706.
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abstract = "Monoclonal antibody 1-12-3 reactive against scup (Stenotomus chrysops) cytochrome P450 E (a teleost CYP IA1) has been used to immunohistochemically localize CYP IA1 within hepatocytes and presumably sinusoidal endothelial and biliary epithelial cells of scup and trout. The goal of the present study was to extend immunohistochemical studies to the ultrastructural level determining intracellular locations of CYP IA1 in fish liver. Juvenile trout (5-10 g) were given i.p. injections once (50 μg/g β-naphthoflavone in cod liver oil; 0.5-ml injectate volume). After 5 days, livers were fixed (0.25{\%} glutaraldehyde) via vascular in situ perfusion, removed, cut in 100-μm slices, infiltrated, and embedded in LR White monomer. Ultrathin sections of exposed livers were incubated in monoclonal antibody 1-12-3, rabbit anti-mouse IgG, and protein G colloidal gold. Membranes of granular endoplasmic reticulum in perinuclear regions of hepatocytes were consistently labeled. In addition, hepatocyte plasma membrane, particularly microvilli at bile canaliculi, was labeled. Biliary epithelial cells were labeled on luminal plasma membrane surrounding biliary passageway. Plasma membrane facing sinusoid and immediately subjacent cytoplasm was labeled in endothelial cells. Presence of CYP IA1 in sinusoidal endothelium could contribute to detoxication and/or bioactivation of blood borne chemicals. Granular endoplasmic reticulum was not uniformly labeled in hepatocytes. Rather, distribution seemed sequestered within highly specific regions and not dispersed along all membrane surfaces. Localization within biliary epithelial cells could signify potential of this cell type to bioactivate polycyclic aromatic hydrocarbons and may explain the common finding of biliary as well as hepatocytic tumors of trout liver.",
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