NADP-dependent dehydrogenases in rat liver parenchyma. I. Methodological studies on the qualitative histochemistry of G6PDH, 6PGDH, malic enzyme and ICDH.
At present soluble NADP-dependent dehydrogenases are histochemically demonstrated in three different ways: according to the standard method incubation in aqueous media leads to the precipitation of formazan, the formation of which depends entirely on the presence of endogeneous NADPH2-tetrazolium reductases. With the two more recently established methods these reductases are by-passed with the use of intermediate electron acceptors incorporated in the medium. In addition, enzyme diffusion is inhibited either by an increased viscosity of the medium (PVA) or by a semipermeable membrane separating the medium from the section. Depending on the technique applied different distribution patterns have been described. By altering the concentrations of substrates, coenzyme, tetrazolium salt and cytochrome oxidase inhibitor, it was possible to improve both the PVA and membrane methods. Although similar results were obtained, because of its advantages the PVA method is recommended in this report and a detailed description is given. Using the latter for the demonstration of glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), malic enzyme (ME) and isocitrate dehydrogenase (ICDH), characteristic distribution patterns were obtained in the liver parenchyma of male and female rats. For the first time a high G6PDH activity could be demonstrated in nonparenchymal cells which are mainly found in zone 1 of the liver acinus.
NADP-dependent dehydrogenases in rat liver parenchyma. I. Methodological studies on the qualitative histochemistry of G6PDH, 6PGDH, malic enzyme and ICDH. At present soluble NADP-dependent dehydrogenases are histochemically demonstrated in three different ways: according to the standard method incubation in aqueous media leads to the precipitation of formazan, the formation of which depends entirely on the presence of endogeneous NADPH2-tetrazolium reductases. With the two more recently established methods these reductases are by-passed with the use of intermediate electron acceptors incorporated in the medium. In addition, enzyme diffusion is inhibited either by an increased viscosity of the medium (PVA) or by a semipermeable membrane separating the medium from the section. Depending on the technique applied different distribution patterns have been described. By altering the concentrations of substrates, coenzyme, tetrazolium salt and cytochrome oxidase inhibitor, it was possible to improve both the PVA and membrane methods. Although similar results were obtained, because of its advantages the PVA method is recommended in this report and a detailed description is given. Using the latter for the demonstration of glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), malic enzyme (ME) and isocitrate dehydrogenase (ICDH), characteristic distribution patterns were obtained in the liver parenchyma of male and female rats. For the first time a high G6PDH activity could be demonstrated in nonparenchymal cells which are mainly found in zone 1 of the liver acinus.