Internucleosomal DNA Cleavage Should not be the Sole Criterion for Identifying Apoptosis

Collins, Russell J.; Harmon, Brian V.; Gobé, Glenda C.; Kerr, Judith

doi:10.1080/09553009214551201

Cited by 431 publications

(196 citation statements)

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“…These tests were positive for both excitotoxic and apoptotic cell death, signifying that both types of cell death degrade DNA into similar fragments that are detected by these methods, but because the two cell death processes are clearly dissimilar by ultrastructural analysis, one must conclude that DNA fragmentation analysis is not a valid or reliable test for diagnosing apoptosis. Our finding that DNA fragmentation tests are not specific for apoptosis and do not reliably distinguish apoptosis from excitotoxic cell death is consistent with similar findings from several other laboratories (34)(35)(36), including a study from the Kerr laboratory (37) [Kerr originally coined the term "apoptosis" (38)]. …”

Section: Distinguishing Excitotoxic From Apoptotic Neuronal Cell Deathsupporting

confidence: 79%

Environmental Agents That Have the Potential to Trigger Massive Apoptotic Neurodegeneration in the Developing Brain

Olney¹,

Farber²,

Wozniak³

et al. 2000

Environmental Health Perspectives

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We review recent findings pertaining to several environmental agents (ethanol, phencyclidine, ketamine, nitrous oxide, barbiturates, benzodiazepines, halothane, isoflurane, and propofol) that have the potential to delete large numbers of neurons from the developing brain by a newly discovered mechanism involving interference in the action of neurotransmitters [glutamate and y-amino butyric acid (GABA) at Nmethyl-D-aspartate (NMDA)1 and GABAA receptors during the synaptogenesis period, also known as the brain growth-spurt period. Transient interference (lasting > 4 hr) in the activity of these transmitters during the synaptogenesis period (the last trimester of pregnancy and the first several years after birth in humans) causes millions of developing neurons to commit suicide (die by apoptosis). Many of these agents are drugs of abuse (ethanol is a prime example) to which the human fetal brain may be exposed during the third trimester by drug-abusing mothers. Ethanol triggers massive apoptotic neurodegeneration in the developing brain by interfering with both the NMDA and GABAA receptor systems, and this can explain the reduced brain mass and lifelong neurobehavioral disturbances associated with intrauterine exposure of the human fetus to ethanol (fetal alcohol syndrome). Exposure of the immature brain in a medical treatment context is also of concern because many of these agents are drugs used frequently as sedatives, tranquilizers, anticonvulsants, or anesthetics in pediatric and/or obstetrical medicine. Because this is a newly discovered mechanism, further research will be required to fully ascertain the nature and degree of risk posed by exposure of the developing human brain to environmental agents that act by this mechanism.

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Section: Distinguishing Excitotoxic From Apoptotic Neuronal Cell Deathsupporting

confidence: 79%

Environmental Agents That Have the Potential to Trigger Massive Apoptotic Neurodegeneration in the Developing Brain

Olney¹,

Farber²,

Wozniak³

et al. 2000

Environmental Health Perspectives

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“…These changes are characteristic of necrotic cell death where DNA fragmentation occurs randomly as a late event secondary to cell death, not necessarily internucleosomal. 29 However, Ramos cells incubated with EPA displayed typical signs of apoptosis. DNA fragmentation was already observed after 12 h (16%) and the TUNEL labeling was intense (Figure 4b).…”

Section: Resultsmentioning

confidence: 99%

Multiplication and death-type of leukemia cell lines exposed to very long-chain polyunsaturated fatty acids

et al. 1998

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Polyunsaturated fatty acids (PUFA) may reduce cell multiplication in cultures of normal, as well as transformed, white blood cells. We assessed the sensitivity of 14 different leukemia cell lines to PUFA by measuring cell number after 3 days of incubation. Ten of the examined cell lines were sensitive to 30, 60 and/or 120 M of arachidonic, eicosapentaenoic and docosahexaenoic acid, whereas four cell lines were resistant. The sensitivity to PUFA was not associated with any particular cell lineage, clinical origin or specific mRNA pattern of bcl-2 and c-myc. Effects on cell viability were assessed by studying cell membrane integrity, DNA fragmentation and cell morphology. The sensitive cell lines Raji and Ramos died by necrosis and apoptosis, respectively, during incubation with eicosapentaenoic acid, whereas the viability of the resistant U-698 cell line was unaffected. The effects of EPA on Raji cells, was counteracted by vitamin E, indicating that lipid peroxidation was involved. However, apoptosis induced by eicosapentaenoic acid in Ramos cells, was unaffected by vitamin E, as well as eicosanoid synthesis inhibitors. In conclusion, our results indicate that a majority of leukemia cell lines are sensitive to PUFA. This sensitivity may be caused by induction of apoptosis or necrosis by very long-chain polyunsaturated fatty acids.

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“…The somewhat confounding data and interpretations derived from these studies may be partially attributable to the criteria used to identify apoptosis. Some of the criteria, such as the size of the nucleus and DNA fragmentation (Collins et al, 1992;Oberhammer et al, 1993;Portera-Cailliau et al, 1995), may not be specific for apoptosis. In our experiments, apoptotic cells were identified as cells exhibiting nuclear fragmentation, a hallmark of apoptosis in different cell types and in response to a variety of stimuli (Lazebnik et al, 1993;Oberhammer et al, 1993).…”

Section: Discussionmentioning

confidence: 99%

Mechanisms of Cell Death Induced by the Mitochondrial Toxin 3-Nitropropionic Acid: Acute Excitotoxic Necrosis and Delayed Apoptosis

1997

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Impaired energy metabolism may play an important role in neuronal cell death after brain ischemia and in late-onset neurodegenerative diseases. Both excitotoxic necrosis and apoptosis have been implicated in cell death induced by metabolic impairment. However, the factors that determine whether cells undergo apoptosis or necrosis are not known. In the present study, metabolic impairment was induced by 3-nitropropionic acid (3-NP), a suicide inhibitor of succinate dehydrogenase. Treatment of cultured rat hippocampal neurons with 3-NP resulted in two types of cell death with distinct morphological, pharmacological, and biochemical features. A rapid necrotic cell death, characterized by cell swelling and nuclear shrinkage, could be completely prevented by the NMDA receptor antagonist MK-801 (10 M) and dose-dependently potentiated by low micromolar levels of extracellular glutamate. A slowly evolving apoptotic death, characterized by nuclear fragmentation, was not attenuated by MK-801 but was prevented by cycloheximide (1 g/ml). The combination of MK-801 and cycloheximide resulted in an almost complete protection against 3-NP-induced cell death. DNA fragmentation, detected by the terminal deoxynucleotidyl transferase-mediated dUTP-X 3Ј nick end-labeling technique, was a late event in apoptosis and also occurred after necrotic cell death. ATP depletion was an early event in the 3-NP-induced neuronal degeneration, and the decline in ATP was exacerbated by glutamate. We conclude that 3-NP triggers two separate cell death pathways: an excitotoxic necrosis as a result of NMDA receptor activation and a delayed apoptosis that is NMDA receptor-independent. Mildly elevated levels of extracellular glutamate shift the cell death mechanism from apoptosis to necrosis.Key words: energy metabolism; succinate dehydrogenase; 3-nitropropionic acid; excitotoxicity; apoptosis; necrosis; nuclear fragmentation; TUNEL; ATP Neuronal function and survival depend on a continuous supply of glucose and oxygen, used to generate ATP through glycolysis and mitochondrial respiration. A perturbation in energy metabolism during conditions such as ischemia, stroke, and brain trauma may cause irreversible neuronal injury. An age-related decline in energy metabolism also may contribute to neuronal loss during normal aging, as well as in neurodegenerative diseases (Wallace, 1994;Beal, 1995).There are discrepancies in the findings regarding the mechanisms of neuronal cell death after metabolic impairment. A large body of evidence supports the "secondary excitotoxicity" hypothesis that a loss of ATP leads to membrane depolarization, removal of the voltage-dependent Mg 2ϩ block of the NMDA receptor, and subsequent activation of NMDA receptor (for review, see Beal, 1992). Both in vivo and in vitro studies have shown that metabolic inhibitors potentiate glutamate and NMDA toxicity (Weller and

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Internucleosomal DNA Cleavage Should not be the Sole Criterion for Identifying Apoptosis

Cited by 431 publications

References 11 publications

Environmental Agents That Have the Potential to Trigger Massive Apoptotic Neurodegeneration in the Developing Brain

Environmental Agents That Have the Potential to Trigger Massive Apoptotic Neurodegeneration in the Developing Brain

Multiplication and death-type of leukemia cell lines exposed to very long-chain polyunsaturated fatty acids

Mechanisms of Cell Death Induced by the Mitochondrial Toxin 3-Nitropropionic Acid: Acute Excitotoxic Necrosis and Delayed Apoptosis

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