Original article
English
Khadija Debri, Mariam Alkhadi, Mabrouk Mohamed, Ali Gurnazi, Aisha Dugani
The National Medical Research Centre, Zawia
LJMR Vol 1, No.1 2002: 64-81
Abstract
The cellular distribution of cytochrome P4503A1 and cytochrome P4503A2 enzymes has been investigated in the hepatic and extrahepatic tissues of the wistar rats by immunocytochemistry using; two highly specific antibodies against rafP4503AI and P4503A2 enzymes. The specificity of these antibodies was verified by a number of techniques; ELISA, western blotting; and immuncadsorbent; assays, {our published study}. Breifly, the previous studies showed that in ELISA assay both antibodies bound strongly to hepatic microsomal fraction from rats treated with pregnenolone 16 a-carbonitrile (PCN). Also, the binding of these antibodies to hepatic and extra hepatic mirosomal fractions from uninduced and induced rats showed differences in expression of cytochrome P450 enzyme (protein) recognized by the two antibodies, providing further clear evidence of antibody specificity (immunoblotting).
In this study we applied immunocytochemistry technique, as an other ????formation investigation for the specificity of each antibody. Distribution and loca1ization of CYP3AI and CYP3A2 in the liver of untreated animals, both CYP3AI and CP3A2 were found to be expressed in the centriobular region. However, some CYP3AI immunoreactivity was also detected in many, but not all, hepatocytes throughout the lobule. However, following treatment of rats with PCN, both CYP3AI Mid CPY3A2 and CPY3A2 were found to be strongly expressed in hepatocytes throughout the lobule, although CYP3A2 showed greater expression in centrilobular region. PCN treatment was also found to result in induction of CYP3AI in specific regions of the small intestine, lung, and kidney. Therefore; the binding of anti-CYP3AI (???IITGS) and anti-CYP3A2 (VTNGA) is exclusive and consistent with specific binding to their target antigens.
Keywords: Immunohistochemical localization of CYP45O3AI & CYP3A2 Enzymes in hepatic and extrahepatic tissues of wistar rat
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