title: Techniques to Distinguish Apoptosis from Necroptosis.
abstract: The processes by which cells die are as tightly regulated as those that govern cell growth and proliferation. Recent studies of the molecular pathways that regulate and execute cell death have uncovered a plethora of signaling cascades that lead to distinct modes of cell death, including "apoptosis," "necrosis," "autophagic cell death," and "mitotic catastrophe." Cells can readily switch from one form of death to another; therefore, it is vital to have the ability to monitor the form of death that cells are undergoing. A number of techniques are available that allow the detection of cell death and when combined with either knockdown approaches or inhibitors of specific signaling pathways, such as caspase or RIP kinase pathways, they allow the rapid dissection of divergent cell death pathways. However, techniques that reveal the end point of cell death cannot reconstruct the sequence of events that have led to death; therefore, they need to be complemented with methods that can distinguish all forms of cell death. Apoptotic cells frequently undergo secondary necrosis under in vitro culture conditions; therefore, novel methods relying on high-throughput time-lapse fluorescence video microscopy are necessary to provide temporal resolution to cell death events. Further, visualizing the assembly of multiprotein signaling hubs that can execute apoptosis or necroptosis helps to explore the underlying processes. Here we introduce a suite of techniques that reliably distinguish necrosis from apoptosis and secondary necrosis, and that enable investigation of signaling platforms capable of instructing apoptosis or necroptosis.
authors.affiliation: Section of Molecular Dermatology, Department of Dermatology, Venereology, and Allergology, Medical Faculty Mannheim, University Heidelberg, 68167 Mannheim, Germany;
authors.forename: Maria
authors.initials: M
authors.lastname: Feoktistova
authors.affiliation: The Breakthrough Toby Robins Breast Cancer Research Centre, Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, United Kingdom;
authors.forename: Fredrik
authors.initials: F
authors.lastname: Wallberg
authors.affiliation: The Breakthrough Toby Robins Breast Cancer Research Centre, Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, United Kingdom;
authors.forename: Tencho
authors.initials: T
authors.lastname: Tenev
authors.affiliation: Section of Molecular Dermatology, Department of Dermatology, Venereology, and Allergology, Medical Faculty Mannheim, University Heidelberg, 68167 Mannheim, Germany;
authors.forename: Peter
authors.initials: P
authors.lastname: Geserick
authors.affiliation: Section of Molecular Dermatology, Department of Dermatology, Venereology, and Allergology, Medical Faculty Mannheim, University Heidelberg, 68167 Mannheim, Germany; Department of Dermatology & Allergology, University Hospital of RWTH Aachen University, 52074 Aachen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Leverkus
authors.affiliation: The Breakthrough Toby Robins Breast Cancer Research Centre, Institute of Cancer Research, Chester Beatty Laboratories, London SW3 6JB, United Kingdom;
authors.forename: Pascal
authors.initials: P
authors.lastname: Meier
mesh_terms: D017209:Apoptosis; D003584:Cytological Techniques / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D018715:Microscopy, Video / Q000379:methods; D009336:Necrosis; D015398:Signal Transduction; D059008:Time-Lapse Imaging / Q000379:methods