Detection and measurement of single-strand breaks in nuclear DNA in fixed lens sections☆,☆☆
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Chromatin collapse during caspase-dependent apoptotic cell death requires DNA fragmentation factor, 40-kDa subunit-/caspase-activated deoxyribonuclease- mediated 3′-OH single-strand DNA breaks
2013, Journal of Biological ChemistryCitation Excerpt :Therefore, the results presented here support the notion that DFF40/CAD acts similarly in cellula, first by introducing single-strand nicks/breaks and later by cleaving the adjacent second strand, yielding laddered patterns of oligonucleosomal-size fragments (see Fig. 10). The presence of single-strand (transient) DNA lesions that permit the rearrangement of genetic material has also been observed during cell differentiation (50–53). More recently, the SSBs generated during terminal differentiation of skeletal muscle cells have been attributed to DFF40/CAD (54, 55).
On the mechanism of organelle degradation in the vertebrate lens
2009, Experimental Eye ResearchCitation Excerpt :Like DNase IIα, DNase IIβ cleaves DNA to produce 3′-phosphoryl/5′-hydroxy ends (Shiokawa and Tanuma, 1999). It has been known for many years that 3′-OH ends rather than 5′-OH ends accumulate during fiber cell denucleation (Modak and Bollum, 1972) so the realization that DNase IIβ is the major nuclease responsible for lens chromatin degradation was surprising. This apparently paradoxical finding may be explained by the presence of endogenous phosphatases, which rapidly convert 3′-PO4 ends generated by DNAse IIβ digestion into 3′-OH ends (De Maria and Bassnett, 2007).
Adult neurogenesis and neuronal regeneration in the brain of teleost fish
2008, Journal of Physiology ParisAdult neurogenesis and neuronal regeneration in the teleost fish brain: Implications for the evolution of a primitive vertebrate trait
2007, Evolution of Nervous SystemsApoptosis in lens development and pathology
2006, DifferentiationChapter 5.6 Cellular and molecular analysis of stress-induced neurodegeneration - methodological considerations
2005, Techniques in the Behavioral and Neural SciencesCitation Excerpt :This approach detects 3′-OH ends of single-stranded DNA after the addition of labeled nucleotides to the open ends of DNA in a procedure known as in situ end-labeling (ISEL). The latter may be achieved using either E. coli polymerase (or its Klenow fragment) in a method called in situ nick-translation (ISNT), or terminal transferase in a method referred to as terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL) (Modak and Bollum, 1972; Gavrieli et al., 1992; Jin et al., 1999). These methods allow the cytochemical demonstration of free DNA strand openings.
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A part of this work was conducted when one us (S. P. M.) was at the Department of Microbiology, University of Kentucky, Lexington, Ky, USA.
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Research was supported jointly by grant 3.466.70 from the Swiss National Fund for Scientific Research and grants CA-08487 and CA-10375 from the National Cancer Institute, USA.