Isolation of Centrolobular and Perilobular Hepatocytes after Phenobarbital Treatment

Daily phenobarbital (PB) injections, on 3-7 consecutive days, induce an intense proliferation of smooth endoplasmic reticulum (ER) associated with a decrease of the glucose-6-phosphatase activity. This situation first affects the centrolobular hepatocytes, enhancing the degree of liver lobule hetero...

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Bibliographic Details
Published in:The Journal of cell biology 1975-07, Vol.66 (1), p.23-41
Main Authors: Wanson, Jean-Claude, Drochmans, Pierre, May, Claude, Penasse, Willy, Popowski, Anna
Format: Article
Language:English
Subjects:
Animals
Cell Separation
Cells
Centrifugation
Centrifugation, Density Gradient
Density
DNA
DNA - analysis
Endoplasmic Reticulum - ultrastructure
Female
Glucose-6-Phosphatase - analysis
Glycogen
Hepatocytes
Histocytochemistry
Injections, Intraperitoneal
Liver
Liver - analysis
Liver - cytology
Liver - ultrastructure
Liver cells
Liver Glycogen - analysis
Microscopy, Electron
Organoids - ultrastructure
Phenobarbital - pharmacology
Phosphorylases - analysis
Proteins - analysis
Rats
RNA
RNA - analysis
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Summary:Daily phenobarbital (PB) injections, on 3-7 consecutive days, induce an intense proliferation of smooth endoplasmic reticulum (ER) associated with a decrease of the glucose-6-phosphatase activity. This situation first affects the centrolobular hepatocytes, enhancing the degree of liver lobule heterogeneity. This experimental model was used for isolation and further subfractionation of hepatocytes on Ficoll density gradients, as described in the preceding paper. Profiles of protein, DNA, RNA, glycogen, phosphorylase, and glucose-6-phosphatase were determined all along the gradient. Two liver cell populations were distinguished: (a) light hepatocytes (mean density 1.10) present the same morphological characteristics as centrolobular cells, i.e., an abundant smooth ER composed of tubular elements, numerous small mitochondria, and few glycogen particles; (b) heavy hepatocytes (mean density 1.14) are characterized by large and compact glycogen areas and prominent rough endoplasmic cisternae, as are the perilobular cells. After incubation in the Wachstein-Meisel medium, centrolobular hepatocytes exhibit dispersed reaction sites of glucose-6-phosphatase activity, whereas perilobular cells present a continuous and intense reaction. Morphometric determinations were carried out for both cell populations. Centrolobular PB hepatocytes are considerably enlarged (mean diameter: 23.7 μm); perilobular hepatocytes have a significantly smaller mean diameter of 19.2 μm, which is close to the values of control liver cells.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.66.1.23