Abstract:
:Neuronal death can be induced by DNA-damaging agents and occurs by apoptosis involving a specific signal-transduction pathway. However, to our knowledge, methods for the quantitative determination of DNA damage in individual neurons have not yet been described. Here we optimize the single-cell gel electrophoresis (SCGE) or "comet"-assay to measure DNA damage within individual neurons growing in dissociated cell culture. In addition, we have written a macro for the NIH Image program to determine the tail moment of individual comets. We have calibrated this method using gamma-irradiated (0-16 Gy) cerebral cortical neurons from the rat central nervous system. Neuronal DNA damage (in the form of DNA strand breaks) occurs in a linear, dose-dependent manner, which can be quantitatively determined in vitro using the SCGE assay. These data demonstrate that the SCGE assay is an effective method to measure DNA damage in individual neurons and may be highly useful for the study of neuronal DNA damage formation, repair and apoptosis.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Morris EJ,Dreixler JC,Cheng KY,Wilson PM,Gin RM,Geller HMdoi
10.2144/99262st02subject
Has Abstractpub_date
1999-02-01 00:00:00pages
282-3, 286-9issue
2eissn
0736-6205issn
1940-9818journal_volume
26pub_type
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