Structural changes of isolated hepatocytes during treatment with digitonin

George A. Cook, Vincent H. Gattone, Andrew P. Evan, Robert A. Harris

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The structural changes accompanying digitonin-induced release of enzymes and metabolites from isolated hepatocytes have been studied by scanning and transmission electron microscopy. In the initial phase, characterized by total release of the cytosolic marker enzyme, lactate dehydrogenase, the plasma membrane was immediately damaged, rapidly followed by extensive damage to the endoplasmic reticulum. The shape of the cell, however, was maintained, and the mitochondria and nucleus remained tightly held together by the cytoskeleton. Mitochondria remained intact initially, whereas the cytosol became less electron dense and the nuclear chromatin was more dispersed. An intermediate phase was characterized by total release of adenylate kinase and most of the glucose-6-phosphatase, marker enzymes for the mitochondrial intermembrane space and the endoplasmic reticulum, respectively. The outer mitochondrial membrane was ruptured, but mitochondria maintained their normal matrix electron density. In the final phase, characterized by the beginning of citrate synthase release from the mitochondrial matrix space, the mitochondria became swollen, and only the nucleus, inner and outer mitochondrial membranes, and the cytoskeleton could be clearly distinguished. Although the plasma membrane could not be readily discerned in electron micrographs after the initial phase, the plasma membrane marker enzyme 5′-nucleotidase remained associated with digitonin-treated hepatocytes. Acetyl-CoA carboxylase was released much more slowly than lactate dehydrogenase, indicating some severe restriction on its release. The release of acetyl-CoA carboxylase closely paralleled the release of glucose-6-phosphatase. The controlled exposure of hepatocytes to digitonin, therefore, leads to the sequential release of soluble, compartmentalized cellular components and some membrane-bound components, but the mitochondrial membrane, cytoskeleton and the nucleoskeleton survive even long-term digitonin treatment.

Original languageEnglish (US)
Pages (from-to)356-367
Number of pages12
JournalBBA - Molecular Cell Research
Volume763
Issue number4
DOIs
StatePublished - Dec 19 1983

Fingerprint

Digitonin
Mitochondrial Membranes
Hepatocytes
Mitochondria
Cytoskeleton
Acetyl-CoA Carboxylase
Glucose-6-Phosphatase
Cell Membrane
Electrons
Enzymes
L-Lactate Dehydrogenase
Endoplasmic Reticulum
Nuclear Matrix
Adenylate Kinase
Citrate (si)-Synthase
5'-Nucleotidase
Scanning Transmission Electron Microscopy
Cell Shape
Cytosol
Chromatin

Keywords

  • (Rat liver)
  • Digitonin effect
  • Electron microscopy
  • Enzyme release
  • Hepatocyte
  • Metabolite release

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Structural changes of isolated hepatocytes during treatment with digitonin. / Cook, George A.; Gattone, Vincent H.; Evan, Andrew P.; Harris, Robert A.

In: BBA - Molecular Cell Research, Vol. 763, No. 4, 19.12.1983, p. 356-367.

Research output: Contribution to journalArticle

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