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Journal of Histochemistry and Cytochemistry, Vol. 48, 963-970, July 2000, Copyright © 2000, The Histochemical Society, Inc.

ARTICLE

Protease-elicited TUNEL Positivity of Non-apoptotic Fixed Cells

István Gála, Tamás Vargaa, Ildikó Szilágyia, Margit Balázsb, József Schlammadingerc, and Gábor Szabó, Jra
a Department of Biophysics and Cell Biology, University Medical School of Debrecen, Debrecen, Hungary
b Department of Hygiene, University Medical School of Debrecen, Debrecen, Hungary
c Department of Human Genetics, University Medical School of Debrecen, Debrecen, Hungary

Correspondence to: Gábor Szabó, Jr, Dept. of Biophysics and Cell Biology, University Medical School of Debrecen, POB 39, 4012 Debrecen, Hungary. E-mail: szabog@jaguar.dote.hu

The appearance of free DNA ends in the chromatin is usually considered an indication of advanced apoptosis. Unexpectedly, the nuclei of non-apoptotic cells derived from mouse thymuses could be specifically labeled by terminal transferase after proteinase K treatment of the fixed, cytocentrifuged samples. Artifactual mechanical or contaminating nucleolytic factors have been ruled out as players in the generation of free DNA ends. The phenomenon was detected in both formaldehyde- and ethanol-fixed specimens, in agarose-embedded fixed cells, and in chromatin spreads. By urea–agarose gel electrophoresis, the average single-strand size of the DNA molecules carrying the free ends was found between 50 and 250 kb. We suggest that ss discontinuities preexisting in the fixed normal cells are unmasked by protease treatment eliciting TUNEL (terminal transferase-mediated nick end-labeling) positivity. (J Histochem Cytochem 48:963–969, 2000)

Key Words: apoptosis, chromatin, fragmentation, protease, TUNEL


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