Germ cell and dose-dependent DNA damage measured by the comet assay in murine spermatozoaa after testicular X-irradiation.
Affiliation
School of Biological Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.Issue Date
2002-09
Metadata
Show full item recordAbstract
The single-cell gel electrophoresis (Comet) assay has been widely used to measure DNA damage in human sperm in a variety of physiological and pathological conditions. We investigated the effects of in vivo radiation, a known genotoxin, on spermatogenic cells of the mouse testis and examined sperm collected from the vas deferens using the neutral Comet assay. Irradiation of differentiating spermatogonia with 0.25-4 Gy X-rays produced a dose-related increase in DNA damage in sperm collected 45 days later. Increases were found when measuring Comet tail length and percentage of tail DNA, but the greatest changes were in tail moment (a product of tail length and tail DNA). Spermatids, spermatocytes, differentiating spermatogonia, and stem cell spermatogonia were also irradiated in vivo with 4 Gy X-rays. DNA damage was indirectly deduced to occur at all stages. The maximum increase was seen in differentiating spermatogonia. DNA damaged cells were, surprisingly, still detected 120 days after stem cell spermatogonia had been irradiated. The distribution of DNA damage among individual sperm cells after irradiation was heterogeneous. This was seen most clearly when changes in the Comet tail length were measured when there were discrete undamaged and damaged populations. After increasing doses of irradiation, an increasing proportion of cells were found in the damaged population. Because a proportion of undamaged sperm cells remains after all but the highest dose, the possibility of normal fertility remains. However, fertilization with a spermatozoa carrying high amounts of DNA damage could lead to effects as diverse as embryonic death and cancer susceptibility in the offspring.Citation
Germ cell and dose-dependent DNA damage measured by the comet assay in murine spermatozoaa after testicular X-irradiation. 2002, 67 (3):854-61 Biol. Reprod.Journal
Biology of ReproductionDOI
10.1095/biolreprod.102.004382PubMed ID
12193394Type
ArticleLanguage
enISSN
0006-3363ae974a485f413a2113503eed53cd6c53
10.1095/biolreprod.102.004382
Scopus Count
Collections
Related articles
- Increased levels of comet-detected spermatozoa DNA damage following in vivo isotopic- or X-irradiation of spermatogonia.
- Authors: Haines GA, Hendry JH, Daniel CP, Morris ID
- Issue date: 2001 Aug 22
- DNA damage in human and mouse spermatozoa after in vitro-irradiation assessed by the comet assay.
- Authors: Haines G, Marples B, Daniel P, Morris I
- Issue date: 1998
- Effect of ATM heterozygosity on heritable DNA damage in mice following paternal F0 germline irradiation.
- Authors: Baulch JE, Li MW, Raabe OG
- Issue date: 2007 Mar 1
- Evaluation of DNA damage in different stages of mouse spermatogenesis after testicular X irradiation.
- Authors: Cordelli E, Fresegna AM, Leter G, Eleuteri P, Spanò M, Villani P
- Issue date: 2003 Oct
- Direct and delayed X-ray-induced DNA damage in male mouse germ cells.
- Authors: Cordelli E, Eleuteri P, Grollino MG, Benassi B, Blandino G, Bartoleschi C, Pardini MC, Di Caprio EV, Spanò M, Pacchierotti F, Villani P
- Issue date: 2012 Jul