Sperm DNA fragmentation is related to sperm morphological staining patterns.
In this prospective comparative study, sperm DNA fragmentation (sDNAfrag) was compared at each step of a sequential semen preparation, with semen parameters according to their degree of severity. At each step (fractions) of the sequential procedure, sDNAfrag was determined: fresh (Raw), after gradient centrifugation, washing, and swim-up (SU) for 70 infertile men enrolled in intracytoplasmic sperm injection cycles. sDNAfrag significantly (P = 0.04; P < 0.0001) decreased throughout the steps of semen preparation, with centrifugation and washing not increasing it. A negative correlation to sperm motility was observed in Raw and SU fractions, and a higher sDNAfrag was observed in samples with lower semen quality. Our results confirm that the steps of the sequential procedure do not compromise sperm DNA integrity and progressively decreased sDNAfrag regardless of the sperm abnormality and that semen parameters with lower quality present higher sDNAfrag. Four distinct patterns were observed, of which the entire sperm head staining was the pattern most expressed in all studied fractions. Additionally, the sperm head gene-rich region staining pattern was reduced by the procedure. This suggests that pattern quantification might be a useful adjunct when performing sDNAfrag testing for male infertility.
Department of Microscopy, Laboratory of Cell Biology, Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Porto, Portugal. Electronic address: rmsa@icbas.up.pt.
Rosália
R
Sá
Centre for Reproductive Genetics Prof. Alberto Barros (CGR-ABarros), 4100-009 Porto, Portugal.
Mariana
M
Cunha
Department of Microscopy, Laboratory of Histology and Embryology, ICBAS-UP, 4050-313 Porto, Portugal.
Eduardo
E
Rocha
Centre for Reproductive Genetics Prof. Alberto Barros (CGR-ABarros), 4100-009 Porto, Portugal; Department of Genetics, Faculty of Medicine, UP, 4200-319 Porto, Portugal.
Alberto
A
Barros
Department of Microscopy, Laboratory of Cell Biology, Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto (UP), Porto, Portugal.
Mário
M
Sousa
D000328:Adult; D053938:DNA Fragmentation; D005260:Female; D006801:Humans; D007248:Infertility, Male / Q000378:metabolism* / Q000473:pathology* / Q000628:therapy; D008297:Male; D008875:Middle Aged; D011446:Prospective Studies; D027724:Reproductive Techniques, Assisted; D055101:Semen Analysis; D013076:Sperm Count; D013077:Sperm Head / Q000378:metabolism / Q000473:pathology; D020554:Sperm Injections, Intracytoplasmic; D013081:Sperm Motility; D013094:Spermatozoa / Q000002:abnormalities* / Q000378:metabolism*; D013194:Staining and Labeling
TUNEL assay; semen parameters; sequential sperm preparation technique; sperm DNA fragmentation; sperm head morphological patterns