Apoptosis and necrosis: Detection, discrimination and phagocytosis

Krysko, Dmitri V.; Berghe, Tom Vanden; D’Herde, Katharina; Vandenabeele, Peter

doi:10.1016/j.ymeth.2007.12.001

Cited by 592 publications

(503 citation statements)

References 61 publications

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“…22 Moreover, PI staining of non-permeabilized-induced cells, which identifies late apoptotic and necrotic cells, remained below 6.5% during the 3 days of observation (Figure 3e), as opposed to 98.5% of control ethanol permeabilized cells (not shown). This shows that a large majority of cells did not die from necrosis.…”

Section: Resultsmentioning

confidence: 87%

Inhibition of active nuclear transport is an intrinsic trigger of programmed cell death in trypanosomatids

et al. 2008

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The link between nucleocytoplasmic transport and apoptosis remains controversial. Nucleocytoplasmic exchange of molecules seems indeed essential for the initiation and execution of the apoptotic programme; but inhibition of nuclear transport factors may also represent a powerful apoptotic trigger. The GTPase Ran (together with its partners), first discovered to be essential in nucleocytoplasmic transport, has multiple key functions in cell biology, and particularly in spindle assembly, kinetochore function and nuclear envelope assembly. Among the Ran partners studied, NTF2 appears to be solely involved in nucleocytoplasmic transport. Here, we localised Ran and several of its partners, RanBP1, CAS and NTF2, at the nuclear membrane in the trypanosomatid Leishmania major. Remarkably, these proteins fused to GFP decorated a perinuclear 'collar' of about 15 dots, colocalising at nuclear pore complexes with the homologue of nucleoporin Sec13. In the other trypanosomatid Trypanosoma brucei, RNAi knockdown of the expression of the corresponding genes resulted in an apoptosis-like phenomenon. These phenotypes show that Ran and its partners have a key function in trypanosomatids like they have in mammals. Our data, notably those about TbNTF2 RNAi, support the idea that active nucleocytoplasmic transport is not essential for the initiation and execution of apoptosis, and, rather, the impairment of this transport constitutes an intrinsic signal for triggering PCD.

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Section: Resultsmentioning

confidence: 87%

Inhibition of active nuclear transport is an intrinsic trigger of programmed cell death in trypanosomatids

et al. 2008

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“…In most cases, their performance is toxic to the cells or require fixation. On a large scale, three major methodologies are currently used in cellular research (Krysko et al 2008;Martinez et al 2010a;Tait 2008):…”

Section: Introductionmentioning

confidence: 99%

Beyond annexin V: fluorescence response of cellular membranes to apoptosis

2012

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Dramatic changes in the structure of cell membranes on apoptosis allow easy, sensitive and non-destructive analysis of this process with the application of fluorescence methods. The strong plasma membrane asymmetry is present in living cells, and its loss on apoptosis is commonly detected with the probes interacting strongly and specifically with phosphatidylserine (PS). This phospholipid becomes exposed to the cell surface, and the application of annexin V labeled with fluorescent dye is presently the most popular tool for its detection. Several methods have been suggested recently that offer important advantages over annexin V assay with the ability to study apoptosis by spectroscopy of cell suspensions, flow cytometry and confocal or twophoton microscopy. The PS exposure marks the integrated changes in the outer leaflet of cell membrane that involve electrostatic potential and hydration, and the attempts are being made to provide direct probing of these changes. This review describes the basic mechanisms underlying the loss of membrane asymmetry during apoptosis and discusses, in comparison with the annexin V-binding assay, the novel fluorescence techniques of detecting apoptosis on cellular membrane level. In more detail we describe the detection method based on smart fluorescent dye F2N12S incorporated into outer leaflet of cell membrane and reporting on apoptotic cell transformation by easily detectable change of the spectral distribution of fluorescent emission. It can be adapted to any assay format.

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“…A large number of intracellular components are released into the intercellular space due to cell burst. Necrosis causes damage to the surrounding tissue because it promotes an inflammatory process (49).…”

Section: Introductionmentioning

confidence: 99%

“…These apoptotic bodies are then phagocyted by surrounding phagocytes, thus preventing the release of the cytoplasmic content to the outside. Hence, the development of an inflammatory response and damage to the neighboring cells are avoided (49,50).…”

Section: Introductionmentioning

confidence: 99%

Cell Death Mechanisms Induced by Cytotoxic Lymphocytes

et al. 2009

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One of the functions of the immune system is to recognize and destroy abnormal or infected cells to maintain homeostasis. This is accomplished by cytotoxic lymphocytes. Cytotoxicity is a highly organized multifactor process. Here, we reviewed the apoptosis pathways induced by the two main cytotoxic lymphocyte subsets, natural killer (NK) cells and CD8 + T cells. In base to recent experimental evidence, we reviewed NK receptors involved in recognition of target-cell, as well as lytic molecules such as perforin, granzymes-A and -B, and granulysin. In addition, we reviewed the Fas-FasL intercellular linkage mediated pathway, and briefly the cross-linking of tumor necrosis factor (TNF) and TNF receptor pathway. We discussed three models of possible molecular interaction between lytic molecules from effector cytotoxic cells and target-cell membrane to induction of apoptosis. Cellular & Molecular Immunology. 2009;6(1):15-25.

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Apoptosis and necrosis: Detection, discrimination and phagocytosis

Cited by 592 publications

References 61 publications

Inhibition of active nuclear transport is an intrinsic trigger of programmed cell death in trypanosomatids

Inhibition of active nuclear transport is an intrinsic trigger of programmed cell death in trypanosomatids

Beyond annexin V: fluorescence response of cellular membranes to apoptosis

Cell Death Mechanisms Induced by Cytotoxic Lymphocytes

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