Induction of mRNA Coding for Phenobarbital-Inducible Form of Microsomal Cytochrome P-450 in Rat Liver by Administration of 1,1-Di(p-Chlorophenyl)-2,2-Dichloroethylene and Phenobarbital1

Abstract

In the preceding paper (Yoshioka, H., et al. (1984) J. Biochem. 95, 937–947), we reported that 1,1-di(p-chlorophenyl)-2,2-dichloro-ethylene (DDE) induced the pheno-barbital (PB)-inducible form of microsomal cytochrome P-450 (P-450(PB-1)) in rat liver. In order to study more precisely the molecular events responsible for the induction of this particular form of cytochrome P-450 by the two chemical compounds, we determined the amounts of the mRNA coding for P-450(PB-1) in the liver of rats given a single dose of PB or DDE. RNA was extracted from the livers of the treated rats and the determination of the specific mRNA was carried out by using the rabbit reticulocyte lysate translation system and by a dot hybridization method using cloned P-450(PB-1) cDNA (Fujii-Kuriyama, Y., et al. (1982) Proc. Natl. Acad. Sci. U.S.79, 2793–2797) as the probe. The amounts of P-450(PB-1) mRNA determined by these two methods at various time points of the induction process showed good agreement. These observations further confirmed the induction of an identical form of cytochrome P-450 by DDE and PB. The maximum level of P-450(PB-1) mRNA, which was about 8-fold higher than the control level, was attained at 20-30 h and at 48–72 h after the administration of PB and DDE, respectively. The mRNA level showed a rapid decrease after the peak in the liver of PB-treated rats, but the decrease was much slower with DDE-treated rats. We conclude that DDE had a more persistent inducing effect on the mRNA level than PB, although these two compounds induced an identical form of cytochrome P-450 in the liver microsomes of the animals.