Presence of double-strand breaks with single-base 3' overhangs in cells undergoing apoptosis but not necrosis.

Didenko, Vladimir V.; Hornsby, Peter J.

doi:10.1083/jcb.135.5.1369

Cited by 179 publications

(168 citation statements)

References 32 publications

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“…The presence of some such 3' -recessed ends in gluco corticoid-treated thymic lymphocytes had previously been deduced by labeling with the Klenow fragment of DNA polymerase (Peitsch et aI., 1993). However, a com pletely opposite conclusion was reached in another re cent study, in which it was suggested that the majority of fragment ends in thymus have single-base 3' overhangs, although some blunt-ended fragments were also present (Didenko and Hornsby, 1996). The methods used to gen erate these latter findings are not quantitative, so the LM-PCR method appears to produce the most reliable information owing to its semiquantitative nature.…”

Section: Klenow Polymerase Treatment (A)mentioning

confidence: 95%

Cerebral Ischemia Produces Laddered DNA Fragments Distinct from Cardiac Ischemia and Archetypal Apoptosis

MacManus

Fliss

Preston³

et al. 1999

J Cereb Blood Flow Metab

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Summary:The electrophoretic pattern of laddered DNA frag ments which has been observed after cerebral ischemia is con sidered to indicate that neurons are dying by apoptosis. Herein the authors directly demonstrate using ligation-mediated poly merase chain reaction methods that 99% of the DNA fragments produced after either global or focal ischemia in adult rats, or produced after hypoxia-ischemia in neonatal rats, have stag gered ends with a 3' recess of approximately 8 to 10 nucleo tides. This is in contrast to archetypal apoptosis in which the DNA fragments are blunt ended as seen during developmental Despite its initial description as long ago as 1980 (Wyllie et aI., 1980), an electrophoretic laddered pattern of DNA fragments remains the premier biochemical hall mark of an apoptotic mode of cell death (Bortner et aI., 1995;Choi, 1996;Walker and Sikorska, 1997). How ever, some exceptions have been noted, for example in neonatal brain after excitotoxicity (van Lookeren Cam pagne et aI., 1995), that prevent the sole use of this criterion as conclusive evidence of apoptosis. This elec trophoretic pattern arises from the cleavage of the linker DNA between nucleosomes in chromatin, resulting in oligomeric fragments of unit size of approximately 180 base pairs (bp). The occurrence of such internucleosomal cleavage has been documented in a wide variety of cells and tissues induced to die by a myriad of insults. The endonuclease responsible for the ordered scission of the DNA backbone remains unknown, but several candidates such as DNase I or II and Nuc 18 have been proposed over the years (Bortner et aI., 1995; Walker and Sikor ska, 1997

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Section: Klenow Polymerase Treatment (A)mentioning

confidence: 95%

Cerebral Ischemia Produces Laddered DNA Fragments Distinct from Cardiac Ischemia and Archetypal Apoptosis

MacManus

Fliss

Preston³

et al. 1999

J Cereb Blood Flow Metab

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“…Doublestranded DNA is subject to the activity of endonucleases, which can induce cuts with staggered ends and blunt ends. 16 The characteristic pattern of apoptosis is double-stranded breaks at internucleosomal DNA sites such that the breaks have staggered ends with 3Ј overhangs comprising 1 or 2 bases, or longer overhangs involving 4 bases. 16 A histochemical method using Taq polymerase detects single-base 3Ј overhangs produced by Ca 2ϩ -dependent DNAase I, whereas the method using TdT detects 1-base 3Ј overhangs as well as multiple-base 3Ј overhangs produced by pH-dependent DNAase II and possibly other DNAases.…”

Section: See P 1422mentioning

confidence: 99%

“…16 The characteristic pattern of apoptosis is double-stranded breaks at internucleosomal DNA sites such that the breaks have staggered ends with 3Ј overhangs comprising 1 or 2 bases, or longer overhangs involving 4 bases. 16 A histochemical method using Taq polymerase detects single-base 3Ј overhangs produced by Ca 2ϩ -dependent DNAase I, whereas the method using TdT detects 1-base 3Ј overhangs as well as multiple-base 3Ј overhangs produced by pH-dependent DNAase II and possibly other DNAases. 16 Although the TdT-based TUNEL assay is less specific, comparable results of the Taq polymerase-based in situ ligation and TdT-based TUNEL methods have been reported in a recent study.…”

Section: See P 1422mentioning

confidence: 99%

“…16 A histochemical method using Taq polymerase detects single-base 3Ј overhangs produced by Ca 2ϩ -dependent DNAase I, whereas the method using TdT detects 1-base 3Ј overhangs as well as multiple-base 3Ј overhangs produced by pH-dependent DNAase II and possibly other DNAases. 16 Although the TdT-based TUNEL assay is less specific, comparable results of the Taq polymerase-based in situ ligation and TdT-based TUNEL methods have been reported in a recent study. 17 In oncocytic necrosis, there is more generalized activation of endonucleases and exonucleases leading to cleavage of nucleosomal and internucleosomal DNA that results in blunt ends of the digested fragments.…”

Section: See P 1422mentioning

confidence: 99%

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Modes of Myocardial Cell Injury and Cell Death in Ischemic Heart Disease

1998

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“…De todas ellas, la más importante es la que se refiere a las alteraciones del ADN. En todo proceso de muerte celular se produce la activación de unas enzimas, las nucleasas, que fragmentan al ADN 3 . Mientras que en la apoptosis se activan selectivamente ciertas endonucleasas que fragmentan al ADN sólo en sitios internucleosomales, en la oncosis se activan más generalizadamente endonucleasas y exonucleasas, que fragmentan al ADN tanto en sitios internucleosomales como nucleosomales.…”

Section: Apoptosis In Cardiovascular Diseasesunclassified

Apoptosis en las enfermedades cardiovasculares

Díez¹

2000

Revista Española de Cardiología

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INTRODUCCIÓNDesde las primeras descripciones clásicas de Virchow se reconocía un tipo de muerte celular: la necrosis. Sin embargo, a finales del siglo pasado, Walter Flemming describió un tipo de muerte celular con características morfológicas diferentes, al que denominó cromatólisis. Tuvo que transcurrir casi un siglo hasta que este tipo de muerte celular fuese adecuadamente caracterizado por Kerr, quien en 1972 propuso el tér-mino apoptosis para denominarlo 1 .En los últimos 20 años, la apoptosis y la necrosis se han considerado como los dos tipos fundamentales de muerte celular. Sin embargo, Majno y Joris han puesto de manifiesto la inconsistencia de este concepto 2 . En efecto, en términos estrictos, la necrosis representa el conjunto de cambios degradativos en los que culmina cualquier tipo de muerte celular. Así, a la necrosis se llega tanto por un proceso de apoptosis, morfológica-mente caracterizado por el arrugamiento y la fragmentación de la célula, como por un proceso de oncosis, en el que la célula se hincha y estalla. Por lo tanto, hoy día cabe distinguir dos tipos fundamentales de muerte celular: el apoptótico y el oncocítico. La apoptosis constituye una forma de muerte celular con características morfológicas y dinámicas distintas de la muerte por oncosis o por necrosis. Desde el punto de vista de la arquitectura celular, la apoptosis equilibra el efecto de la proliferación celular. Por ello, las alteraciones de la regulación de la apoptosis pueden participar en el desarrollo de numerosas enfermedades. Es el caso de la apoptosis exagerada, que conduce a la atrofia y al fallo de la función de un órgano, o de la apoptosis insuficiente, que propicia el remodelado estructural organotisular. En los últimos años se asiste a una gran expansión en la investigación sobre apoptosis, una investigación en la que también participa la comunidad cardiovascular. Desde muy diversos ámbitos de la medicina cardiovascular se han aportado observaciones sobre el posible papel de la apoptosis en síndromes que oscilan desde los defectos de conducción hasta la insuficiencia cardíaca congestiva, desde la aterosclerosis hasta los aneurismas. No hay duda de que esas observaciones puntuales acabarán configurando conceptos etiopatogénicos y fisiopatológi-cos que serán la base del diseño de nuevas estrategias diagnósticas y terapéuticas. A R T Í C U L O D E R E V I S I Ó N

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Presence of double-strand breaks with single-base 3' overhangs in cells undergoing apoptosis but not necrosis.

Cited by 179 publications

References 32 publications

Cerebral Ischemia Produces Laddered DNA Fragments Distinct from Cardiac Ischemia and Archetypal Apoptosis

Cerebral Ischemia Produces Laddered DNA Fragments Distinct from Cardiac Ischemia and Archetypal Apoptosis

Modes of Myocardial Cell Injury and Cell Death in Ischemic Heart Disease

Apoptosis en las enfermedades cardiovasculares

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