Reactive Oxygen Species and Cryopreservation Promote DNA Fragmentation in Equine Spermatozoa

Julie Baumber, Barry A. Ball, Jennifer J. Linfor, Stuart A Meyers

Research output: Contribution to journalArticle

215 Citations (Scopus)

Abstract

The objective of this study was to examine the effect of reactive oxygen species (ROS) and cryopreservation on DNA fragmentation of equine spermatozoa. In experiment 1, equine spermatozoa were incubated (1 hour, 38°C) according to the following treatments: 1) sperm alone; 2) sperm + xanthine (X, 0.3 mM)-xanthine oxidase (XO, 0.025 U/mL); 3) sperm + X (0.6 mM)-XO (0.05 U/roL); and 4) sperm + X (1 mM)-XO (0.1 U/mL). In experiment 2, spermatozoa were incubated (1 hour, 38°C) with X (1 mM)-XO (0.1 U/mL) and either catalase (200 U/mL), superoxide dismutase (SOD, 200 U/mL), or reduced glutathione (GSH, 10 mM). Following incubation, DNA fragmentation was determined by the single cell gel electrophoresis (comet) assay. In experiment 3, equine spermatozoa were cryopreserved, and DNA fragmentation was determined in fresh, processed, and postthaw sperm samples. In experiment 1, incubation of equine spermatozoa in the presence of ROS, generated by the X-XO system, increased DNA fragmentation (P < .005). In Experiment 2, the increase in DNA fragmentation associated with X-XO treatment was counteracted by the addition of catalase and GSH but not by SOD, suggesting that hydrogen peroxide and not superoxide appears to be the ROS responsible for such damage. In experiment 3, cryopreservation of equine spermatozoa was associated with an increase (P < .01) in DNA fragmentation when compared with fresh or processed samples. This study indicates that ROS and cryopreservation promote DNA fragmentation in equine spermatozoa; the involvement of ROS in cryopreservation-induced DNA damage remains to be determined.

Original languageEnglish (US)
Pages (from-to)621-628
Number of pages8
JournalJournal of Andrology
Volume24
Issue number4
StatePublished - Jul 2003

Fingerprint

Cryopreservation
DNA Fragmentation
Horses
Spermatozoa
Reactive Oxygen Species
Comet Assay
Catalase
Xanthine
Xanthine Oxidase
Superoxides
Hydrogen Peroxide
DNA Damage
Superoxide Dismutase
Glutathione

Keywords

  • Antioxidants
  • DNA damage
  • Horse
  • Oxidative stress
  • Sperm

ASJC Scopus subject areas

  • Endocrinology

Cite this

Reactive Oxygen Species and Cryopreservation Promote DNA Fragmentation in Equine Spermatozoa. / Baumber, Julie; Ball, Barry A.; Linfor, Jennifer J.; Meyers, Stuart A.

In: Journal of Andrology, Vol. 24, No. 4, 07.2003, p. 621-628.

Research output: Contribution to journalArticle

Baumber, Julie ; Ball, Barry A. ; Linfor, Jennifer J. ; Meyers, Stuart A. / Reactive Oxygen Species and Cryopreservation Promote DNA Fragmentation in Equine Spermatozoa. In: Journal of Andrology. 2003 ; Vol. 24, No. 4. pp. 621-628.
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