Abstract
Apoptosis and necrosis represent two different mechanisms by which cells die. The dynamics of cellular lesions in these two processes differ. In particular we demonstrate that plasma membrane damage, occurring as a primary event during necrosis represents, on the contrary, a delayed but massive phenomenon during apoptosis. In consequence there are different kinetics of propidium iodide incorporation by necrotic and apoptotic thymocytes. This represents the basis for the flow cytometric identification of different cellular subsets. Analysis of these subsets after sorting showed that clearly apoptotic cells, which are not able to exclude propidium iodide for long incubation periods, do not show any morphologically detectable membrane damage. The kinetics of propidium iodide incorporation in vivo in isolated rat thymocytes can therefore be used in flow cytometric analysis. This technique can be used instead of DNA staining of ethanol-treated cells or nick translation to recognize apoptotic cells, and distinguish apoptosis from necrosis, without killing the cell.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arends MJ, Morris RG, Wyllie AH (1990) Apoptosis. The role of the endonuclease. Am J Pathol 136:593–608
Barbieri D, Troiano L, Grassilli E, Agnesini C, Cristofalo EA, Monti D, Capri M, Cossarizza A, Franceschi C (1992) Inhibition of apoptosis by zinc: a reappraisal. Biochem Biophys Res Commun 187:1256–1261
Cohen JJ (1991) Programmed cell death in the immune system. Adv Immunol 50:55–85
Collins RJ, Harmon BV, Gobè GC, Kerr JFR (1992) Internucleosomal DNA cleavage should not be the sole criterion for identifying apoptosis. Int Radiat Biol 61:451–453
Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hots MA, Lassota P, Traganos F (1992) Features of apoptotic cells measured by flow cytometry. Cytometry 13:795–808
Ellis RE, Yuan J, Horvitz HR (1991) Mechanisms and functions of cell death. Annu Rev Cell Biol 7:663–698
Falcieri E, Martelli AM, Bareggi R, Cataldi A, Cocco L (1993) The protein kinase inhibitor staurosporin induces morphological changes typical of apoptosis in Molt-4 cells without concomitant DNA fragmentation. Biochem Biophys Res Commun 193:19–25)
Gerschenson LE, Rotello RJ (1992) Apoptosis: a different type of cell death. FASEB J 6:2450–2455
Giannakis C, Forbes IJ, Zalewski PD (1991) Ca2+/Mg2+-dependent nuclease: tissue distribution, relationship to inter-nucleosomal DNA fragmentation and inhibition by Zn2+. Biochem Biophys Res Commun 181:915–920
Kerr JFR, Wyllie AH, Curie AR (1972) Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26:239–257
Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271–279
Sen S, D'Incalci M (1992) Apoptosis. Biochemical events and relevance to cancer chemotherapy. FEBS Lett 307:122–127
Swat W, Ignatowicz L, Kisielow P (1991) Detection of apoptosis of immature CD4+8+ thymocytes by flow cytometry. J Immunol Methods 137:79–87
Telford WG, King LE, Fraker PJ (1992) Comparative evaluation of several DNA binding dyes in the detection of apoptosis-associated chromatin degradation by flow cytometry. Cytometry 13:137–143
William JT (1991) Programmed cell death: apoptosis and oncogenes. Cell 65:1097–1098
Wyllie AH, Kerr JFR, Currie AR (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68:251–306
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Vitale, M., Zamai, L., Mazzotti, G. et al. Differential kinetics of propidium iodide uptake in apoptotic and necrotic thymocytes. Histochemistry 100, 223–229 (1993). https://doi.org/10.1007/BF00269095
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00269095