Distribution of Organelles and Membranes between Hepatocytes and Nonhepatocytes in a Stereological Study

When biochemical studies on the liver are interpreted, the cells of the sinusoidal area frequently receive little attention because, compared to hepatocytes, their contribution to subcellular fractions is assumed insignificant. A systematic stereological analysis of liver parenchyma was therefore performed in order to determine the distribution of organelles and membranes between hepatocytic and nonhepatocytic cells, namely endothelial, Kupffer, and fat-storing cells. The livers were fixed by vascular perfusion and the data were corrected for systematic errors dur to section thickness and compression. The extracellular space compartment includes the lumina of sinusoids (10.6%), the space of Disse (4.9%), and the bile canaliculi (0.4%). Hepatocytes constitute 78% of parenchymal volume; the nonhepatocytes account for 6.3% and consist of 2.8% endothelial cells, 2.1% Kupffer cells, and 1.4% fat- storing cells. The nonhepatocytes contribute 55% of the volume of lipid droplets in the liver, 43% of the lysosomes, and 1.2% of the mitochondria. Although the nonhepatocytes account for only 8% of the total surface area of parenchymal membranes, they contain 26.5% of all the plasma membranes, 32.4% of the lysosomal membranes, 15.1% of the Golgi apparatus 6.4% of the endoplasmic reticulum, and 2.4% of the mitochondrial membranes. The data demonstrate the extent to which nonhepatocytic organelles can potentially contaminate subcellular fractions used for biochemical studies. Particularly important for the interpretation of studies on lysosomes, plasma membrane, and Golgi apparatus is the finding that an appreciable part of these organelles may be derived from cell types other than hepatocytes.

[1]  P. Siekevitz,et al.  GOLGI FRACTIONS PREPARED FROM RAT LIVER HOMOGENATES , 1973, The Journal of cell biology.

[2]  A. Loud A QUANTITATIVE STEREOLOGICAL DESCRIPTION OF THE ULTRASTRUCTURE OF NORMAL RAT LIVER PARENCHYMAL CELLS , 1968, The Journal of cell biology.

[3]  P. Siekevitz,et al.  Golgi fractions prepared from rat liver homogenates. I. Isolation procedure and morphological characterization. , 1973 .

[4]  K. Wake "Sternzellen" in the liver: perisinusoidal cells with special reference to storage of vitamin A. , 1971, The American journal of anatomy.

[5]  Hans Rudolf Gnägi,et al.  CORRELATED MORPHOMETRIC AND BIOCHEMICAL STUDIES ON THE LIVER CELL , 1969, The Journal of cell biology.

[6]  P. Siekevitz,et al.  GOLGI FRACTIONS PREPARED FROM RAT LIVER HOMOGENATES , 1973, The Journal of cell biology.

[7]  E. Weibel Stereological principles for morphometry in electron microscopic cytology. , 1969, International review of cytology.

[8]  O. Greengard,et al.  CYTOMORPHOMETRY OF DEVELOPING RAT LIVER AND ITS APPLICATION TO ENZYMIC DIFFERENTIATION , 1972, The Journal of cell biology.

[9]  G. Paumgartner,et al.  Uptake of bile acids by perfused rat liver. , 1976, The American journal of physiology.

[10]  D. Bendall,et al.  Tissue fractionation studies. 13. Analysis of mitochondrial fractions from rat liver by density-gradient centrifuging. , 1959, The Biochemical journal.

[11]  E. Wisse,et al.  An ultrastructural characterization of the endothelial cell in the rat liver sinusoid under normal and various experimental conditions, as a contribution to the distinction between endothelial and Kupffer cells. , 1972, Journal of ultrastructure research.

[12]  K. Wake Distribution of Vitamin A in the Liver , 1964 .

[13]  W. Scherle,et al.  A simple method for volumetry of organs in quantitative stereology. , 1970, Mikroskopie.

[14]  E. Weibel,et al.  Similarity of supporting tissue in fish gills and the mammalian reticuloendothelium. , 1972, Journal of ultrastructure research.

[15]  G. Palade,et al.  CELL JUNCTIONS IN AMPHIBIAN SKIN , 1965, The Journal of cell biology.

[16]  E. Reynolds THE USE OF LEAD CITRATE AT HIGH pH AS AN ELECTRON-OPAQUE STAIN IN ELECTRON MICROSCOPY , 1963, The Journal of cell biology.

[17]  H. Popper,et al.  Fat-storing cells (lipocytes) in human liver. , 1966, Archives of pathology.

[18]  E. Wisse Kupffer cell reactions in rat liver under various conditions as observed in the electron microscope. , 1974, Journal of ultrastructure research.

[19]  John H. Luft,et al.  IMPROVEMENTS IN EPOXY RESIN EMBEDDING METHODS , 1961, The Journal of biophysical and biochemical cytology.

[20]  E. Wisse,et al.  An electron microscopic study of the fenestrated endothelial lining of rat liver sinusoids. , 1970, Journal of ultrastructure research.

[21]  A. Novikoff,et al.  LOCALIZATION OF ACID PHOSPHATASE ACTIVITY IN HEPATIC LYSOSOMES BY MEANS OF ELECTRON MICROSCOPY , 1961, The Journal of biophysical and biochemical cytology.

[22]  R. E. Miles Estimating aggregate and overall characteristics from thick sections by transmission microscopy , 1976 .

[23]  Andreas Schaefer,et al.  Current capabilities and limitations of available stereological techniques , 1972 .

[24]  E. Weibel,et al.  A MORPHOMETRIC STUDY OF THE REMOVAL OF PHENOBARBITAL-INDUCED MEMBRANES FROM HEPATOCYTES AFTER CESSATION OF TREATMENT , 1973, The Journal of cell biology.

[25]  A. Amar‐Costesec,et al.  ELECTRON MICROSCOPE EXAMINATION OF SUBCELLULAR FRACTIONS III. Quantitative Analysis of the Microsomal Fraction Isolated from Rat Liver , 1971 .

[26]  Toshio Ito,et al.  Über die Kupfferschen Sternzellen und die “Fettspeicherungszellen” (“fat storing cells”) in der Blutkapillarenwand der menschlichen Leber. , 1952 .

[27]  F. G. Zaki Principles and Techniques of Electron Microscopy , 1975 .

[28]  Ewald R. Weibel,et al.  CORRELATED MORPHOMETRIC AND BIOCHEMICAL STUDIES ON THE LIVER CELL , 1969, The Journal of cell biology.

[29]  B. Benacerraf,et al.  A HISTOCHEMICAL STUDY OF ACID AND ALKALINE PHOSPHATASE IN MOUSE LIVERS DURING VARIOUS CONDITIONS MODIFYING ACTIVITY OF THE RETICULOENDOTHELIAL SYSTEM , 1961, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[30]  S. Shibasaki,et al.  Electron microscopic study on the hepatic sinusoidal wall and the fat-storing cells in the normal human liver. , 1968, Archivum histologicum Japonicum = Nihon soshikigaku kiroku.

[31]  D. Mizuno,et al.  Possible heterogeneity of the distribution of lysosomal marker enzymes among "lysosomal particles" of rat liver. , 1972, Biochimica et biophysica acta.

[32]  K. Wake DEVELOPMENT OF VITAMIN A-RICH LIPID DROPLETS IN MULTIVESICULAR BODIES OF RAT LIVER STELLATE CELLS , 1974, The Journal of cell biology.

[33]  E. Wisse Observations on the fine structure and peroxidase cytochemistry of normal rat liver Kupffer cells. , 1974, Journal of ultrastructure research.

[34]  Ewald R. Weibel,et al.  An improved apparatus for perfusion fixation with automatic pressure control , 1971, Journal of microscopy.

[35]  T. Berg,et al.  Distribution of lysosomal enzymes between parenchymal and Kupffer cells of rat liver. , 1973, Biochimica et biophysica acta.