Mito-TEMPO improves preservation of cooled canine semen over a 72-hour period of storage

Document Type : Full paper (Original article)

Authors

1 Graduated from Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Theriogenology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

10.22099/ijvr.2026.52842.7905

Abstract

Background: The positive effect of Mito-TEMPO, as a mitochondria-targeted antioxidant, on semen preservation has been reported in various species; however, little information is available on canine semen in this regard. Aims: The present study was conducted to evaluate the effect of Mito-TEMPO on preservation of cooled canine semen over a 72 h period of storage. Methods: In study I, the effect of 0 (as control), 25, 50 and 100 μM concentrations of Mito-TEMPO were evaluated on sperm motility, morphology, viability and plasma membrane integrity at time points 0, 24, 48 and 72 h. In study II, the effect of 0 (as control) and 100 μM (as the most effective dose recognized in study I) concentration of Mito-TEMPO was assessed on reactive oxygen species (ROS) and mitochondrial membrane potential at time points 0 and 72 h. Results: In study I, motility, velocity, viability, and proportion of sperm with normal morphology and intact plasma membrane decreased in all groups over time (P<0.05), but this decrease was slower in sperm treated with different doses of Mito-TEMPO than untreated sperm (P<0.05). In this regard, decrease in total and progressive motility observed from 24 h onward in the untreated group; however, such decrease was detected form 48 h onward in groups treated with different doses of Mito-TEMPO (P<0.05). Although all doses of Mito-TEMPO were effective, treatment with 100 μM concentration of Mito-TEMPO provided the best preservation as compared with the control group (P<0.05). Furthermore, study II revealed that ROS generation increased and mitochondrial membrane potential decreased over time in both groups (P<0.05); however, treatment with 100 μM concentration of Mito-TEMPO ameliorated the increase in ROS and the decrease in mitochondrial membrane potential over time (P<0.05). Conclusion: Mito-TEMPO enhances structural and functional characteristics of canine sperm preserved at 4°C for 72 h, and could serve as an effective antioxidant in canine semen extender.

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References

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