Quantitative analysis of oxidized guanine, 8-oxoguanine, in mitochondrial DNA by immunofluorescence method
- PMID: 19513676
- DOI: 10.1007/978-1-59745-521-3_13
Quantitative analysis of oxidized guanine, 8-oxoguanine, in mitochondrial DNA by immunofluorescence method
Abstract
8-Oxoguanine (8-oxoG), an oxidized form of guanine, is one of the major mutagenic lesions generated under oxidative stress. Oxidative damage in mitochondrial DNA has been implicated as a causative factor for a wide variety of degenerative diseases as well as for cancer during aging. We established a quantitative method for in situ detection of 8-oxoG in mitochondrial DNA in a single-cell level using a monoclonal antibody. Specific detection of 8-oxoG in mitochondrial DNA was confirmed by pre-treatment of samples with DNase I or MutM, the latter excising 8-oxoG opposite C in DNA. We then analyzed 8-oxoG dynamics in mitochondrial DNA of the wild-type and 8-oxoG DNA glycosylase (OGG1)-deficient mouse cells after exposure to hydrogen peroxide. Intensities for the 8-oxoG immunoreactivity in mitochondrial DNA were increased immediately after the exposure to hydrogen peroxide in both types of cells. The increased intensities returned to basal levels within a few hours only in wild-type cells, but not in OGG1-deficient cells which exhibited the increased intensities even 24 h after the exposure. These results indicate that OGG1 is a major enzyme for excision repair of 8-oxoG in mitochondrial DNA in mouse cells, and that our method described here is appropriate to study 8-oxoG dynamics in mitochondrial DNA.
Similar articles
-
Construction and characterization of a cell line deficient in repair of mitochondrial, but not nuclear, oxidative DNA damage.Methods Mol Biol. 2009;554:251-64. doi: 10.1007/978-1-59745-521-3_16. Methods Mol Biol. 2009. PMID: 19513679
-
Functional cooperation of Ogg1 and Mutyh in preventing G: C-->T: a transversions in mice.Fukuoka Igaku Zasshi. 2004 Jan;95(1):17-30. Fukuoka Igaku Zasshi. 2004. PMID: 15031996
-
Mitochondrial repair of 8-oxoguanine is deficient in Cockayne syndrome group B.Oncogene. 2002 Dec 12;21(57):8675-82. doi: 10.1038/sj.onc.1205994. Oncogene. 2002. PMID: 12483520
-
Different DNA repair strategies to combat the threat from 8-oxoguanine.Mutat Res. 2007 Jan 3;614(1-2):69-76. doi: 10.1016/j.mrfmmm.2006.03.007. Mutat Res. 2007. PMID: 16769088 Review.
-
Oxidative damage in nucleic acids and Parkinson's disease.J Neurosci Res. 2007 Apr;85(5):919-34. doi: 10.1002/jnr.21191. J Neurosci Res. 2007. PMID: 17279544 Review.
Cited by
-
Endothelial Endothelin Receptor A Expression Is Associated With Podocyte Injury and Oxidative Stress in Patients With Focal Segmental Glomerulosclerosis.Kidney Int Rep. 2021 Apr 22;6(7):1939-1948. doi: 10.1016/j.ekir.2021.04.013. eCollection 2021 Jul. Kidney Int Rep. 2021. PMID: 34307988 Free PMC article.
-
Particle coatings but not silver ions mediate genotoxicity of ingested silver nanoparticles in a mouse model.NanoImpact. 2017 Jan;5:92-100. doi: 10.1016/j.impact.2017.01.003. Epub 2017 Jan 26. NanoImpact. 2017. PMID: 28944309 Free PMC article.
-
Biomarkers of nucleic acid oxidation - A summary state-of-the-art.Redox Biol. 2021 Jun;42:101872. doi: 10.1016/j.redox.2021.101872. Epub 2021 Jan 28. Redox Biol. 2021. PMID: 33579665 Free PMC article.
-
POLB: A new role of DNA polymerase beta in mitochondrial base excision repair.DNA Repair (Amst). 2017 Dec;60:A1-A5. doi: 10.1016/j.dnarep.2017.11.002. Epub 2017 Nov 6. DNA Repair (Amst). 2017. PMID: 29129598 Free PMC article. Review.
-
Neurotoxicity of cytarabine (Ara-C) in dorsal root ganglion neurons originates from impediment of mtDNA synthesis and compromise of mitochondrial function.Free Radic Biol Med. 2018 Jun;121:9-19. doi: 10.1016/j.freeradbiomed.2018.04.570. Epub 2018 Apr 23. Free Radic Biol Med. 2018. PMID: 29698743 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Research Materials
Miscellaneous
