Measurement of NADPH-ferrihemoprotein reductase content in sections of liver.

Abstract

We developed a method for measuring the content of NADPH-ferrihemoprotein reductase in sections of liver. First, reductase in sections of rat liver was detected with the indirect immunoperoxidase reaction. Subsequently, specific absorbances were measured in the stained sections by microphotometry. Then, the resulting specific absorbances were converted into the reductase content in the sections using an apparent extinction coefficient obtained from a nitrocellulose binding assay. The average of the reductase content in hepatocytes in periportal, intermediate, and perivenous zones thus measured was consistent with the value in liver homogenates estimated by enzyme-linked immunosorbent assay. Therefore, the present method gave accurate measurement of the reductase content in the sections. Perivenous hepatocytes contained 1.5 times as much reductase (1.15 nmol/g liver, mean for five animals) as that in periportal hepatocytes (0.74 nmol/g liver). The reductase content in hepatocytes in the intermediate zone (0.93 nmol/g liver) was intermediate between values of the periportal and perivenous hepatocytes.