Susceptibility of human sperm to in situ DNA denaturation is strongly correlated with DNA strand breaks identified by single-cell electrophoresis.
Susceptibility of mammalian sperm DNA to low pH- or heat-induced denaturation in situ has shown very strong dose-response relationships with animal and human exposure to chemical and physical toxicants and also fertility potential. In this study, 23 human semen samples representing a wide range in percentage (7-86%) of sperm exhibiting abnormally high susceptibility of DNA in situ to denaturation were studied for the integrity of their DNA using alkaline comet assay (single-cell microgel electrophoresis, pH 10.0). The percentage of comets observed for these samples ranged from 5 to 95%; these data correlated strongly with the percentage of sperm with increased DNA denaturability (r = 0.973; P < 0.001). Labeling of 3' ends of nicked DNA sites with 5-bromo-2'-deoxyuridine 5'-triphosphate (BrdUTP) followed by tagging with FITC-BrdUTP monoclonal antibody and flow cytometry also indicated significantly strong correlations of BrdUTP incorporation with both abnormal susceptibility of DNA to denaturation (r = 0.859, P < 0.001) and comet assay (r = 0.812, P < 0.001). The relationship among susceptibility of sperm chromatin to acid denaturation in situ, BrdUTP incorporation, and formation of comets suggests that DNA fragmentation monitored by these assays may have important physiological relevance in terms of sperm quality and fertility potential.
Aravindan
G R
GR
Olson Biochemistry Laboratories, South Dakota State University, Brookings 57007, USA.
Bjordahl
J
J
Jost
L K
LK
Evenson
D P
DP
D000143:Acids / Q000494:pharmacology; D000468:Alkalies / Q000494:pharmacology; D001710:Biotin; D003037:Cobalt Radioisotopes; D053938:DNA Fragmentation; D003856:Deoxyuracil Nucleotides; D004307:Dose-Response Relationship, Radiation; D004586:Electrophoresis / Q000379:methods; D005434:Flow Cytometry; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D013094:Spermatozoa / Q000187:drug effects / Q000473:pathology* / Q000528:radiation effects; D013194:Staining and Labeling
D000143:Acids; D000468:Alkalies; D003037:Cobalt Radioisotopes; D003856:Deoxyuracil Nucleotides; D001710:Biotin