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PMID: 8918817 Published · ppublish English Journal Article

A new approach for the electrophoretic detection of apoptosis.

Nucleic acids research ·Vol. 24 ·No. 20 ·1996-10-15 ·Pages 4092-3

Eldadah BA, Yakovlev AG, Faden AI

Abstract

Apoptotic cell death is often characterized by internucleosomal cleavage of genomic DNA, which exhibits a distinctive ladder upon electrophoresis. However, techniques used for the isolation and detection of DNA to demonstrate laddering may not be sufficiently sensitive, particularly when cleaved DNA is present at modest levels. We propose a new approach for isolating total cellular DNA using a silica-based resin that improves the resolution of DNA laddering. In addition, we introduce a rapid DNA labeling method that can increase the sensitivity of detecting DNA laddering. Each of these methods can be used for DNA from cell cultures or tissues.

MeSH Terms
Animals Apoptosis Brain/metabolism Cells, Cultured Cerebellum/metabolism DNA/analysis,metabolism DNA Fragmentation DNA-Directed DNA Polymerase/metabolism Electrophoresis, Agar Gel/methods Nucleosomes/metabolism Rats Taq Polymerase
Chemicals
Nucleosomes DNA Taq Polymerase DNA-Directed DNA Polymerase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Eldadah B A
Department of Neurology and the Georgetown Institute for Cognitive and Computational Sciences, Georgetown University Medical Center, Washington, DC 20007, USA.
Yakovlev A G
Faden A I
References (16)
16 references, click to expand
  1. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics.
    Br J Cancer. 1972 Aug;26(4):239-57 PMID: 4561027
  2. Removal of serum from primary cultures of cerebellar granule neurons induces oxidative stress and DNA fragmentation: protection with antioxidants and glutamate receptor antagonists.
    J Neurosci Res. 1996 Feb 15;43(4):465-75 PMID: 8699532
  3. Gangliosides prevent glutamate and kainate neurotoxicity in primary neuronal cultures of neonatal rat cerebellum and cortex.
    Proc Natl Acad Sci U S A. 1988 Oct;85(19):7351-5 PMID: 2902628
  4. Novel non-templated nucleotide addition reactions catalyzed by procaryotic and eucaryotic DNA polymerases.
    Nucleic Acids Res. 1988 Oct 25;16(20):9677-86 PMID: 2460825
  5. Traumatic brain injury in the rat: characterization of a lateral fluid-percussion model.
    Neuroscience. 1989;28(1):233-44 PMID: 2761692
  6. A simple and rapid method for detection of apoptosis in human cells.
    Nucleic Acids Res. 1992 Oct 11;20(19):5243 PMID: 1408849
  7. Microscale autoradiographic method for the qualitative and quantitative analysis of apoptotic DNA fragmentation.
    J Cell Physiol. 1993 Mar;154(3):519-26 PMID: 8436601
  8. Apoptosis: the biochemistry and molecular biology of programmed cell death.
    Endocr Rev. 1993 Apr;14(2):133-51 PMID: 8325248
  9. Induction of apoptosis in cerebellar granule neurons by low potassium: inhibition of death by insulin-like growth factor I and cAMP.
    Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):10989-93 PMID: 8248201
  10. Neuroprotective effects of 619C89, a use-dependent sodium channel blocker, in rat traumatic brain injury.
    Brain Res. 1995 Feb 27;673(1):133-40 PMID: 7757466
  11. Mechanisms of delayed cell death following hypoxic-ischemic injury in the immature rat: evidence for apoptosis during selective neuronal loss.
    Brain Res Mol Brain Res. 1995 Mar;29(1):1-14 PMID: 7769986
  12. The use of C0t-1 probe DNA for the detection of low levels of DNA fragmentation.
    Biochem Cell Biol. 1994 Nov-Dec;72(11-12):649-53 PMID: 7654339
  13. Activation of G proteins bidirectionally affects apoptosis of cultured cerebellar granule neurons.
    J Neurochem. 1995 Dec;65(6):2425-31 PMID: 7595536
  14. Evidence of apoptotic cell death after experimental traumatic brain injury in the rat.
    Am J Pathol. 1995 Dec;147(6):1575-83 PMID: 7495282
  15. Molecular biology of CNS injury.
    J Neurotrauma. 1995 Oct;12(5):767-77 PMID: 8594206
  16. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation.
    Nature. 1980 Apr 10;284(5756):555-6 PMID: 6245367
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1996-10-15
Pages
4092-3
Language
English
Region
England
NLM ID
0411011
PMCID
PMC146208
Subset
IM
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