Germ cell and dose-dependent DNA damage measured by the comet assay in murine spermatozoaa after testicular X-irradiation.

2.50
Hdl Handle:
http://hdl.handle.net/10541/83582
Title:
Germ cell and dose-dependent DNA damage measured by the comet assay in murine spermatozoaa after testicular X-irradiation.
Authors:
Haines, Grant A; Hendry, Jolyon H; Daniel, C Paul; Morris, Ian D
Abstract:
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.
Affiliation:
School of Biological Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.
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 Reproduction
Issue Date:
Sep-2002
URI:
http://hdl.handle.net/10541/83582
DOI:
10.1095/biolreprod.102.004382
PubMed ID:
12193394
Type:
Article
Language:
en
ISSN:
0006-3363
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorHaines, Grant A-
dc.contributor.authorHendry, Jolyon H-
dc.contributor.authorDaniel, C Paul-
dc.contributor.authorMorris, Ian D-
dc.date.accessioned2009-10-05T16:05:08Z-
dc.date.available2009-10-05T16:05:08Z-
dc.date.issued2002-09-
dc.identifier.citationGerm 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.en
dc.identifier.issn0006-3363-
dc.identifier.pmid12193394-
dc.identifier.doi10.1095/biolreprod.102.004382-
dc.identifier.urihttp://hdl.handle.net/10541/83582-
dc.description.abstractThe 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.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshDNA-
dc.subject.meshDNA Damage-
dc.subject.meshDose-Response Relationship, Radiation-
dc.subject.meshElectrophoresis-
dc.subject.meshMale-
dc.subject.meshMice-
dc.subject.meshSpermatozoa-
dc.subject.meshTestis-
dc.subject.meshX-Rays-
dc.titleGerm cell and dose-dependent DNA damage measured by the comet assay in murine spermatozoaa after testicular X-irradiation.en
dc.typeArticleen
dc.contributor.departmentSchool of Biological Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.en
dc.identifier.journalBiology of Reproductionen
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