DNA integrity in sexed bull sperm assessed by neutral Comet assay and sperm chromatin structure assay

Research output: Contribution to journalArticlepeer-review

Author(s)

  • G B Boe-Hansen
  • I D Morris
  • A K Ersboll
  • T Greve
  • P Christensen

Department/unit(s)

Publication details

JournalTheriogenology
DatePublished - 1 Apr 2005
Issue number6
Volume63
Number of pages14
Pages (from-to)1789-1802
Original languageEnglish

Abstract

During the production of sex-sorted spermatozoa from bull semen, the cells are exposed to a number of potential hazards including: dilution, centrifugation, incubation, exposure to DNA stains and laser light. These factors may affect the survival capacity and fertilization potential of the sperm. The objective of this study was to determine whether sex-sorted bull spermatozoa have more DNA damage than sperm from conventional processed bull semen. Two methods were used 0 to determine DNA integrity: the neutral Comet assay (NCA) and the sperm chromatin structure assay (SCSA). The NCA showed that the conventional samples had a higher tail moment (TM) (P < 0.017) than the sorted samples and that there was no difference between the samples in tail length (TL) (P = 0.36). The SCSA showed that the DNA fragmentation index (DFI) was higher for conventional than the sorted samples (P = 0.011), but the standard deviation of DFI (SD-DFI) was higher for the sorted samples (P < 0.001). We conclude that the NCA and SCSA can be used in assessing DNA integrity in bovine sperm and that cell sorting by flow cytometry improves the integrity of the sperm cell population. Additionally the results from the SCSA indicated that the sex-sorted sperm had less homogenous sperm chromatin. In the future assessment of sperm DNA integrity may be used to select bulls for sperm sex sorting and optimizing sperm sex sorting procedures. (c) 2004 Elsevier Inc. All rights reserved.

    Research areas

  • flow cytometry, single cell gel electrophoresis, sex sorting spermatozoa, bovine, DNA, CELL GEL-ELECTROPHORESIS, IN-VITRO FERTILIZATION, HUMAN SPERMATOZOA, SEMEN QUALITY, MAMMALIAN SPERM, FIELD FERTILITY, FLOW-CYTOMETRY, BOVINE SPERM, DAMAGE, SUSCEPTIBILITY

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