Application of enzymehistochemical methods to isolated subcellular fractions and to sucrose-ficoll density gradients. A contribution to the comparison of histochemical and biochemical data

Histochemistry. 1977 Aug 1;53(2):165-81. doi: 10.1007/BF00498491.

Abstract

To compare histochemical and biochemical determinations of enzyme activities, enzymehistochemical procedures are applied to sections of pellets of subcelluar fractions. These investigations are of value to determine the subcellular localization of histochemically demonstrable enzyme activities and to test the homogeneity of an isolated fraction. In homogenating duckling liver a great part of the endothelial cells is not destructed and consequently is found in the nuclear fraction. Kupffer cell lysosomes land in the heavy mitochondrial fraction, whereas hepatocyte lysosomes are chiefly found in the light mitochondrial fraction. beta-Glucuronidase activity shows a preferentially microsomal localization. Application of enzymehistochemical staining reactions to discontinuous gradients and comparison with biochemical data provides additional information about the validity of an enzymehistochemical reaction. In rat liver the tetrazolium reductases show a distinctly dual localization: activity in the mitochondrial band and in microsomal bands. As to their localization in different bands of the gradients non-specific esterases demonstrate a clear pH-dependency.

Publication types

  • Comparative Study

MeSH terms

  • Acetylglucosaminidase / analysis
  • Acid Phosphatase / analysis
  • Alkaline Phosphatase / analysis
  • Animals
  • Cell Fractionation / methods
  • Centrifugation, Density Gradient / methods
  • Ducks
  • Ficoll
  • Glucose-6-Phosphatase / analysis
  • Glucuronidase / analysis
  • Histocytochemistry
  • Leucyl Aminopeptidase / analysis
  • Liver / enzymology*
  • Liver / ultrastructure
  • Male
  • Microscopy, Electron
  • NADH Tetrazolium Reductase / analysis
  • Nucleotidases / analysis
  • Subcellular Fractions / enzymology*
  • Sucrose

Substances

  • Ficoll
  • Sucrose
  • NADH Tetrazolium Reductase
  • Nucleotidases
  • Alkaline Phosphatase
  • Acid Phosphatase
  • Glucose-6-Phosphatase
  • Glucuronidase
  • Acetylglucosaminidase
  • Leucyl Aminopeptidase