Abstract
Apoptosis and necrosis reflect the program of cell death employed by a dying cell and the final stage of death, respectively. Whereas apoptosis is defined as a physiological, highly organized cell death process, necrosis is commonly considered to be accidental and uncontrolled. Physiological and weak pathological death stimuli preferentially induce apoptosis, while harsh non-physiological insults often immediately instigate (primary) necrosis. If an apoptosing cell transits into a phase of plasma membrane disintegration, this stage of death is referred to as secondary or post-apoptotic necrosis.
Here, we present several conditions that stimulate primary and/or secondary necrosis and show that necrosis displays considerably different time courses. For subclassification of necrotic phenotypes we employed a flow cytometric single-tube 4-color staining technique including annexin A5-FITC, propidium iodide, DiIC1(5), and Hoechst 33342.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ellis RE, Yuan JY, Horvitz HR (1991) Mechanisms and functions of cell death. Annu Rev Cell Biol 7:663–698
Van Cruchten S, Van Den Broeck W (2002) Morphological and biochemical aspects of apoptosis, oncosis and necrosis. Anat Histol Embryol 31:214–223
Majno G, Joris I (1995) Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 146:3–15
Munoz LE, Lauber K, Schiller M et al (2010) The role of defective clearance of apoptotic cells in systemic autoimmunity. Nat Rev Rheumatol 6:280–289
Janko C, Schorn C, Grossmayer GE et al (2008) Inflammatory clearance of apoptotic remnants in systemic lupus erythematosus (SLE). Autoimmun Rev 8:9–12
Walker NI, Harmon BV, Gobe GC et al (1988) Patterns of cell death. Methods Achiev Exp Pathol 13:18–54
Nicotera P, Leist M, Manzo L (1999) Neuronal cell death: a demise with different shapes. Trends Pharmacol Sci 20:46–51
Zong WX, Thompson CB (2006) Necrotic death as a cell fate. Genes Dev 20:1–15
Golstein P, Kroemer G (2007) Cell death by necrosis: towards a molecular definition. Trends Biochem Sci 32:37–43
Raffray M, Cohen GM (1997) Apoptosis and necrosis in toxicology: a continuum or distinct modes of cell death? Pharmacol Ther 75:153–177
Davies KJ (1999) The broad spectrum of responses to oxidants in proliferating cells: a new paradigm for oxidative stress. IUBMB Life 48:41–47
Burniston JG, Chester N, Clark WA et al (2005) Dose-dependent apoptotic and necrotic myocyte death induced by the beta2-adrenergic receptor agonist, clenbuterol. Muscle Nerve 32:767–774
Munoz LE, Maueröder C, Chaurio R, Berens C, Herrmann M, Janko C (2013) Colourful death: Six-parameter classification of cell death by flow cytometry-Dead cells tell tales. Autoimmunity. [Epub ahead of print]
Elstein KH, Zucker RM (1994) Comparison of cellular and nuclear flow cytometric techniques for discriminating apoptotic subpopulations. Exp Cell Res 211:322–331
Vermes I, Haanen C, Steffens-Nakken H et al (1995) A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods 184:39–51
Scarlett JL, Sheard PW, Hughes G et al (2000) Changes in mitochondrial membrane potential during staurosporine-induced apoptosis in Jurkat cells. FEBS Lett 475:267–272
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
Acknowledgments
This work was supported by SFB643 TP B5, by the Emerging Fields Initiative (EFI) of the FAU Erlangen-Nuremberg, by the K & R Wucherpfennigstiftung, by the Interdisciplinary Center of Clinical Research (IZKF) at the University Hospital Erlangen-Nuremberg, and by the training Grant GK SFB 643 from the German Research Community (DFG).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Janko, C. et al. (2013). Navigation to the Graveyard-Induction of Various Pathways of Necrosis and Their Classification by Flow Cytometry. In: McCall, K., Klein, C. (eds) Necrosis. Methods in Molecular Biology, vol 1004. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-383-1_1
Download citation
DOI: https://doi.org/10.1007/978-1-62703-383-1_1
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-382-4
Online ISBN: 978-1-62703-383-1
eBook Packages: Springer Protocols