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Effects of Alpha-G Rutin Supplementation in Sperm Freezing Extender on Dog Sperm Cryopreservation

  • Park, Sang-Hyoun (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jeon, Yubyeol (Department of Theriogenology and Reproductive Biotechnology, College of Veterinary Medicine and Bio-Safety Research Institute, Jeonbuk National University) ;
  • Talha, Nabeel Abdelbagi Hamad (Department of Theriogenology and Reproductive Biotechnology, College of Veterinary Medicine and Bio-Safety Research Institute, Jeonbuk National University) ;
  • Yu, Il-Jeoung (Department of Theriogenology and Reproductive Biotechnology, College of Veterinary Medicine and Bio-Safety Research Institute, Jeonbuk National University)
  • Received : 2019.07.23
  • Accepted : 2019.10.08
  • Published : 2019.10.31

Abstract

This study was designed to investigate the effects of alpha-glucosyl rutin (G-rutin) and its comparative effects with other antioxidants (glutathione: GSH, catalase: CATA and beta-mercaptoethanol : ${\beta}ME$) on dog sperm freezing. In the first experiment (E1), the spermatozoa were diluted in freezing extender supplemented with 0 (control), 0.001, 0.01, or 0.1% G-rutin and frozen using liquid nitrogen ($LN_2$). The progressive motility, reactive oxygen species (ROS) level and apoptosis of spermatozoa were assessed after sperm thawing at $37^{\circ}C$ for 25 sec. In the second experiment (E2), 0.1% G-rutin group was compared with 10 mM ${\beta}-ME$, $5{\mu}M$ GSH and $50{\mu}M$ CATA groups by assaying progressive motility, viability and gene expression of Bcl-2 and SMCP after sperm freezing and thawing. In E1, 0.1% G-rutin group showed higher (P < 0.05) post-thaw progressive motility and lower (P < 0.05) ROS levels. In E2, the expressions of SMCP in G-rutin group were higher (P < 0.05) than in CATA group while Bcl-2 expression of G-rutin group was higher (P < 0.05) than ${\beta}-ME$ and CATA groups. However, there were no significant differences in progressive motility and viability. Therefore, we suggest that G-rutin can be used as a potentially antioxidative supplement in dog sperm freezing extender on the basis of gene expression related to motility and apoptosis as well as ROS level.

Keywords

References

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