NADPH-generating system: influence on microsomal mono-oxygenase stability during incubation for the liver-microsomal assay with rat and mouse S9 fractions

Mutat Res. 1984 Dec;129(3):281-97. doi: 10.1016/0027-5107(84)90083-6.

Abstract

Activity levels of 7-ethoxycoumarin O-deethylase (ED), aminopyrine N-demethylase (APD), p-nitroanisole O-demethylase (p-NAD) and glucose-6-phosphate dehydrogenase (G-6-PDH) were determined in incubation mixtures for the liver-microsomal assay (LMA) at time 0 and after 1 and 2 h incubation under conditions for mutagenic assay. The experiments were performed with S9 liver fractions from mice (induced with Na-phenobarbital and beta-naphthoflavone) and rats (induced with Aroclor 1254) with and without G-6-PDH in the incubation mixtures. In the absence of G-6-PDH the activities were significantly lower at time 0 in the mouse. The pattern of stability, however, was similar for the activities, with an increase of stability after 1 and 2 h of pre-incubation (an exception for p-NAD). Only ED activity showed a similar behaviour in the rat. No differences were present for APD and p-NAD activities at time 0 in the rat, but the enzyme stabilities were significantly decreased after 2 h of incubation (about 15% and 10% for APD and p-NAD respectively) in the absence of G-6-PDH. At time 0, the amounts of G-6-PDH differed between mouse and rat fractions; however, during the incubations for LMA they decreased by about 57% and 53% for the two species, respectively. In addition to the above biochemical results, the presence of exogenous G-6-PDH in the incubations for the mutagenic assay, significantly increased the mitotic gene conversion and mitotic crossing-over of dimethylnitrosamine (DMN) and AR2MNFN (a nitroimidazo[2,1-b]thiazole) in the D7 strain of Saccharomyces cerevisiae.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 7-Alkoxycoumarin O-Dealkylase
  • Aminopyrine N-Demethylase / metabolism
  • Animals
  • Biotransformation
  • Crossing Over, Genetic / drug effects
  • Female
  • Glucosephosphate Dehydrogenase / metabolism
  • Kinetics
  • Mice
  • Microsomes, Liver / enzymology*
  • Mixed Function Oxygenases / metabolism*
  • Mutagenicity Tests
  • Mutagens / pharmacology*
  • Mutation*
  • NADP / metabolism*
  • Oxidation-Reduction
  • Oxidoreductases, O-Demethylating / metabolism
  • Oxygenases / metabolism
  • Rats
  • Rats, Inbred Strains
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics

Substances

  • Mutagens
  • NADP
  • 4-nitroanisole O-demethylase
  • Mixed Function Oxygenases
  • Oxidoreductases, O-Demethylating
  • Glucosephosphate Dehydrogenase
  • Oxygenases
  • 7-Alkoxycoumarin O-Dealkylase
  • Aminopyrine N-Demethylase