Immunocytochemical study of cytochrome P450 (1A1/1A2) induction in murine ocular tissues

Abstract

C57BL/6 and DBA/2 mice were injected intraperitoneally with beta-naphthoflavone in corn oil and killed 48 hr later. Control animals received an injection of corn oil. The immunoreactivity of cytochrome P450 1A1/1A2 expressed in different ocular tissues and liver was examined with goat anti-P450 antibody (primary antibody) and gold-conjugated anti-goat antibody (secondary antibody). DBA/2 mice, which are non-responsive to aryl hydrocarbon treatment, showed negligible levels of immunoreactivity toward anti-P450 1A1/1A2 antibody in all ocular tissues, whether or not the animals were treated with beta-naphthoflavone. In responsive C57BL/6 mice, however, the immunoreactivity of the uveal tissues, especially ciliary non-pigmented epithelium, was markedly increased by beta-naphthoflavone treatment. The time course of induction of P450 1A1/1A2 immunoreactivity was very similar for the liver and ciliary non-pigmented epithelium, although the maximum level of immunoreactivity of the ciliary epithelium reached in 48 hr after inducer treatment was about 25% of that of liver. The present results support our previous observations that the P4501A enzyme activities (e.g. aryl hydrocarbon hydroxylase) in the liver and eye of C57BL/6 mice are under the same genetic regulation. Further, this study is the first demonstration of P450 isoform induction in specific ocular tissues of the whole animal.