Phenobarbital inducibility and differences in protein expression of an animal model

Aldehyde dehydrogenases (ALDHs) are a group of enzymes which catalyze the conversion of aldehydes to the corresponding carboxylic acids in a NAD(P)(+)-dependent reaction. In mammals, different ALDHs are constitutively expressed in liver, stomach, eye and skin. In addition, inducible ALDH-isoenzymes are detectable in many tissues; apart from other physico- and immuno-chemical differences, two cytosolic ALDHs (ALDH1A3 and ALDH3A1) are known to be activated in rat liver, by different types of inducers of drug metabolism. Phenobarbital-type inducers increase the ALDH1A3, while polycyclic hydrocarbons (such as BaP and TCDD) increase the expression of the two members of ALDH3A subfamily (3A1 and 3A2). In this study, we used two Wistar rat substrains which have been well-characterized for different inducibility of ALDH1A3 enzyme activity after treatment with phenobarbital. Animals that respond (RR) or do not respond (rr) to treatment have been inbred for almost 25 years, offering a useful experimental model. Apart from the level of ALDH1A3 induced enzyme expression after phenobarbital treatment, no other differences between the two substrains have been noticed, as far as drug metabolizing enzyme activities (like the pentoxy- and ethoxy-O-dealkylation rate) are concerned. According to the present results, the ALDH1A3 expression is still the only difference between the two substrains. Immunoblotting experiments with polyclonal antibodies raised against CYP2B1 or/and CYP1A1/1A2 showed no differences between the two substrains. Additionally, data concerning time- and dose-response induction of ALDH1A3 after phenobarbital and griseofulvin treatment are presented. It is concluded that these two Wistar rat substrains represent a unique animal model for studying what seems to be the only difference between these substrains - the genetic basis of the phenobarbital induction.