Immunohistochemical Localization and Differential Expression of Cytochrome P450 3A27 in the Gastrointestinal Tract of Rainbow Trout

• Published in: Toxicology and applied pharmacology 2001-12, Vol.177 (2), p.94-102
• Main Authors: Lee, Su-Jun, Hedstrom, Olaf R., Fischer, Kay, Wang-Buhler, Jun-Lan, Sen, Alaattin, Cok, Ismet, Buhler, Donald R.
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Aryl Hydrocarbon Hydroxylases; Biological and medical sciences; Blotting, Northern; Blotting, Western; Cecum - enzymology; CYP3A; CYP3A27; CYP3A27 protein; Cytochrome P-450 CYP3A; Cytochrome P-450 Enzyme System - biosynthesis; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; Digestive System - enzymology; Digestive System - metabolism; Enzymes and enzyme inhibitors; Female; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Enzymologic; General aspects. Methods; Hydroxylation; immunohistochemical localization; Immunohistochemistry; intestine; Isoenzymes - biosynthesis; Isoenzymes - genetics; Isoenzymes - metabolism; Male; Medical sciences; Microsomes, Liver - enzymology; Oncorhynchus mykiss; Oncorhynchus mykiss - metabolism; Oxidoreductases; Oxidoreductases, N-Demethylating - biosynthesis; Oxidoreductases, N-Demethylating - genetics; Oxidoreductases, N-Demethylating - metabolism; Progesterone - metabolism; rainbow trout; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Testosterone - metabolism; Toxicology
• ISSN: 0041-008X; 1096-0333
• DOI: 10.1006/taap.2001.9289

In mammals the cytochrome P450 3A (CYP3A) subfamily isoforms are primarily expressed in liver and intestines with lesser amounts found in other tissues. The aim of this study was to examine the cellular localization and the expression pattern of CYP3A27 in the gastrointestinal tract (GI tract) of a freshwater teleost species, the rainbow trout (Oncorhynchus mykiss), a fish model used extensively for toxicological and carcinogenesis research. Using an avidin biotinylated enzyme complex and 3,3′-diaminobenzidine staining, strong cytoplasmic immunohistochemical staining was observed for CYP3A27 protein in hepatocytes and in enterocytes of the intestinal ceca and the proximal descending intestine when probed with a polyclonal antibody raised against rainbow trout P450 LMC5, a CYP3A protein. The intensity of epithelial staining decreased distally along the GI tract with faint staining observed in the epithelial cells examined near the vent. Western blot analysis was supportive of the immunohistochemistry results. Northern blot analysis also demonstrated that CYP3A27 mRNA was expressed along the entire GI tract. The major area of CYP3A27 mRNA expression was in the intestinal ceca, followed by the proximal descending intestine, at levels that were about three- to five-fold and two- to four-fold, respectively, greater than seen in the liver of the fish studied. Monooxygenase activities of intestinal ceca microsomes against testosterone and progesterone confirmed the presence of active CYP3A enzyme in this tissue. These results suggest that the intestine of rainbow trout may possesses substantial capacity for first-pass metabolism of xenobiotics by CYP3A27, which makes it an excellent model in which to study the consequence of such metabolism.