Fertility and flow cytometry evaluations of ram frozen semen in plant-based extender supplemented with Mito-TEMPO

Nader Asadzadeh; Zahra Abdollahi; Saeid Esmaeilkhanian; Reza Masoudi

doi:10.1016/j.anireprosci.2021.106836

Fertility and flow cytometry evaluations of ram frozen semen in plant-based extender supplemented with Mito-TEMPO

Anim Reprod Sci. 2021 Aug 29:233:106836. doi: 10.1016/j.anireprosci.2021.106836. Online ahead of print.

Authors

Nader Asadzadeh¹, Zahra Abdollahi², Saeid Esmaeilkhanian¹, Reza Masoudi³

Affiliations

¹ Animal Science Research Institute of Iran (ASRI), Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran.
² Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran.
³ Animal Science Research Institute of Iran (ASRI), Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran. Electronic address: rezamasoudi@ut.ac.ir.

PMID: 34500193
DOI: 10.1016/j.anireprosci.2021.106836

Abstract

Semen cryopreservation is an effective strategy for distributing spermatozoa for artificial insemination, but this process reduces the fertility potential of post-thawed spermatozoa. The current study was conducted to assess effects of the novel mitochondria-targeted antioxidant "Mito-TEMPO" on ram sperm quality and fertility potential during the freeze-thawing process in a plant-based extender. Semen samples were diluted in extenders supplemented with 0, 0.5, 5, 50 and 500 µM Mito-TEMPO and then frozen using a standard protocol. Motility, abnormal morphology, acrosome integrity, membrane integrity, mitochondria membrane potential, viability, apoptotic-like changes, lipid peroxidation, DNA fragmentation, H₂O₂ concentration and fertility potential were assessed after thawing. Results indicated that with the 5 and 50 μM Mito-TEMPO there was a greater (P ≤ 0.05) percentage of sperm total motility, progressive motility, acrosome integrity and viability as well as less (P ≤ 0.05) lipid peroxidation and late apoptotic-like changes. Membrane integrity and mitochondria membrane potential were greater (P ≤ 0.05) with the 50 μM Mito-TEMPO extender supplementation. Furthermore, with 0.5, 5 and 50 μM Mito-TEMPO supplementations there was a greater (P ≤ 0.05) average path velocity and lesser (P ≤ 0.05) percentages of spermatozoa with DNA fragmentation and relatively greater H₂O₂ concentration. Results from the fertility experiment indicated the 5 and 50 μM Mito-TEMPO treatments resulted in greater pregnancy, parturition and lambing rates. It, therefore, is concluded that Mito-TEMPO may enhance quality and fertility potential of post-thawed semen of rams.

Keywords: Antioxidant; Cryopreservation; Fertility; Mito-TEMPO; Sheep.