Heterogeneous expression of phenobarbital-inducible cytochrome P-450 genes within the hepatic acinus in the rat

Abstract

Within the hepatic acinus, the functional unit of liver parenchyma, the induction of cytochrome P-450 protein by phenobarbital is manifested primarily in hepatocytes located closer to the hepatic venule, i.e., distal hepatocytes. The objective of this study was to determine the levels of cytochromes P-450b and P-450e mRNAs in populations of hepatocytes originating in the proximal or distal half of the liver acinus in the rat, as an approach to the elucidation of the mechanisms responsible for the heterogeneous zonal expression of cytochrome P-450 protein. The development of a new method to isolate hepatocytes originating from the proximal or distal half of the liver acinus enabled the measurement of total cytochrome P-450 content and of cytochromes P-450b and P-450e mRNAs in these hepatocytes. Levels of cytochromes P-450b and P-450e mRNAs were assessed in proximal and distal hepatocytes by Northern blot hybridization of poly(A+)RNA with a cDNA recognizing sequences of these two cytochromes. The kinetics of induction were defined by measuring these parameters after a single phenobarbital injection. Cytochrome P-450 mRNA levels reached maximum induction at 16 hr, returning to basal values by 48 hr. In contrast, total cytochrome P-450 microsomal protein content reached maximum induction after 33 hr. Hepatocytes of the distal half of the hepatic acinus responded to phenobarbital with higher levels of cytochromes P-450b and P-450e mRNAs than proximal hepatocytes. These results indicated that there is modulation of the expression of the cytochromes P-450b and P-450e genes within the hepatic acinus.