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Journal of Histochemistry and Cytochemistry, Vol. 46, 1175-1184, October 1998, Copyright © 1998, The Histochemical Society, Inc.

ARTICLE

Pre-apoptotic Alterations in Hepatocytes of TNF{alpha}-treated Galactosamine-sensitized Mice

Sabine Angermüllera, Gerald Künstleb, and Gisa Tiegsc
a Department of Anatomy and Cell Biology II, University of Heidelberg, Heidelberg, Germany
b Biochemical Pharmacology, University of Konstanz, Konstanz, Germany
c Institute of Experimental and Clinical Pharmacology and Toxicology, University of Erlangen-Nürnberg, Erlanger, Germany

Correspondence to: Sabine Angermüller, Dept. of Anatomy and Cell Biology II, U. of Heidelberg, Im Neuenheimer Feld 307, D-69120 Heidelberg, Germany..

Tumor necrosis factor (TNF) induces apoptotic death of hepatocytes in the galactosamine (GalN)-sensitized mouse liver after 5 hr. In our study, the most remarkable sign of the early stage of apoptosis was the focal rupture of the outer mitochondrial membrane. Parts of the inner membrane extended through the gap of the outer membrane, whereas the rest of the inner membrane still formed the cristae. This feature appeared in hepatocytes before chromatin condensation. With the diaminobenzidine technique for localization of cytochrome oxidase activity, the reaction product was detectable by light and electron microscopy. Ten percent of the hepatocytes were apoptotic, with condensed chromatin and high enzyme activity, 37% were pre-apoptotic, without chromatin condensation but high enzyme activity, and 53% had neither condensed chromatin nor a remarkable reaction product of cytochrome oxidase activity. Fas (APO-1, CD95) molecules on the plasma membrane of hepatocytes increased and were represented immunohistochemically in cells without chromatin condensation. DNA strand breaks were also detectable before chromatin aggregation. The results of this study indicate that mitochondria play a pivotal role in pre-apoptotic hepatocytes, together with an increase of the Fas molecule on the plasma membrane and with the occurrence of DNA strand breaks in the nucleus. (J Histochem Cytochem 46:1175–1183, 1998)

Key Words: hepatocytes, apoptosis, mitochondria, cytochrome oxidase, Fas, DNA strand breaks


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