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Journal of Histochemistry and Cytochemistry, Vol. 50, 1599-1609, December 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

NF{kappa}B and Caspase-3 Activity in Apoptotic Hepatocytes of Galactosamine-sensitized Mice Treated with TNF{alpha}

Dan Tapalagaa, Gisa Tiegsb, and Sabine Angermüllera
a Department of Anatomy and Cell Biology II, University of Heidelberg, Heidelberg, Germany
b Institute of Experimental and Clinical Pharmacology and Toxicology, University of Erlangen-Nürnberg, Nürnberg, Germany

Correspondence to: Sabine Angermüller, Dept. of Anatomy and Cell Biology II, U. of Heidelberg, Im Neuenheimer Feld 307, D-69120 Heidelberg, Germany. E-mail: Sabine.Angermueller@urz.uni-heidelberg.de

Tumor necrosis factor-{alpha} (TNF{alpha}) induces apoptosis in hepatocytes only under transcriptional arrest induced by galactosamine (GalN). In this study we demonstrated the shuttle of the transcription factor NF{kappa}B (nuclear factor-kappa B) in the liver tissue of mice within 30 min–4.5 hr hours after GalN/TNF{alpha} treatment. NF{kappa}B translocation from cytoplasm to the nucleus is initiated by its separation from the inhibitory I{kappa}B proteins which include I{kappa}B{alpha}, I{kappa}Bß, and I{kappa}B. Thirty minutes after GalN/TNF{alpha} administration, NF{kappa}Bp65 in hepatocellular nuclei becomes increasingly detectable and reaches its highest level after 2.5 hr. Then export back into cytoplasm begins but, surprisingly, approximately 30% of NF{kappa}B remains in the nuclear fraction and appears as an immunoprecipitate in the nuclei of apoptotic hepatocytes. Non-apoptotic hepatocytes do not show any reaction product in the nuclei 4.5 hr after treatment. Correspondingly, the amount of dissociated I{kappa}Bß decreases in the cytoplasm up to 2.5 hr and increases again afterwards, although it does not reach the level of the control samples. No evidence of I{kappa}Bß in the nuclei was found either immunocytochemically or biochemically. Caspase-3 activity, which is responsible for apoptosis, increases significantly after 3.5 hr. At that time, apoptotic hepatocytes can occasionally be observed and, 4.5 hr after GalN/TNF{alpha} treatment, constitute approximately 30% of the hepatocytes.

(J Histochem Cytochem 50:1599–1609, 2002)

Key Words: NF{kappa}B, I{kappa}Bß, caspase-3 activity, apoptosis, hepatocytes, GalN-sensitized TNF{alpha}-treated, mice


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