X-ray induced DNA double-strand breaks in human sperm.
A methodology for quantifying DNA double-strand breaks in human sperm is described. Sperm from three healthy human donors on three separate days each were irradiated with 12.5, 25, 50 and 100 cGy X-rays. Linear dose-response effects were observed in migrated DNA from sperm nuclei when electrophoresed under neutral conditions. RNase and proteinase K treatments for longer duration were necessary, to decondense the chromatin and presumably to release the broken DNA for migration in the electrophoretic field in a dose-dependent manner. An increase in DNA migration was observed with as low as 12.5 cGy, but damage was observed in all samples at 25 cGy. No evidence of repair of these X-ray-induced DNA double-strand breaks was observed during a 2 h period.
Singh
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Department of Bioengineering, School of Medicine, University of Washington, Seattle 98195, USA. narendra@u.washington.edu
Stephens
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D000328:Adult; D019457:Chromosome Breakage / Q000235:genetics*; D004247:DNA / Q000187:drug effects / Q000528:radiation effects*; D004249:DNA Damage; D004260:DNA Repair / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D019286:Endopeptidase K / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000009:adverse effects; D008297:Male; D012260:Ribonucleases / Q000494:pharmacology; D013094:Spermatozoa / Q000187:drug effects / Q000528:radiation effects*; D014965:X-Rays / Q000009:adverse effects
D004247:DNA; D006861:Hydrogen Peroxide; D012260:Ribonucleases; D019286:Endopeptidase K