Preservation of the quality and fertility potential of post-thawed rooster sperm using MitoQ.

Cryopreservation of rooster spermatozoa is an efficient procedure to spread qualified semen samples for reproductive goals in commercial flocks, but the freeze-thawing process reduces the quality and fertility potential of post-thawed sperm cells. This study was aimed to investigate the effect of the mitochondria-targeted antioxidant MitoQ on rooster sperm quality and fertility potential preservation during freeze-thawing process. Semen samples were collected and diluted in the Lake medium, assigned into five equal aliquots, supplemented with 0, 1, 10, 100 and 1000 nM MitoQ, and cryopreserved in liquid nitrogen. After thawing, sperm motility, membrane functionality, abnormal morphology, mitochondria active potential, acrosome integrity, viability, apoptosis status, lipid peroxidation, DNA fragmentation, ROS concentration and fertility potential were evaluated. According to the results, freezing extender supplementation with 10 and 100 nM MitoQ presented higher (P ≤ 0.05) total motility, progressive motility, average path velocity, membrane functionality, mitochondria active potential, acrosome integrity and viability compared to the other groups. On the other hand, lipid peroxidation, apoptosis rate, DNA fragmentation and ROS concentration were lower (P ≤ 0.05) in groups received 10 and 100 nM MitoQ compared to other groups. Moreover, fertility rate was higher in groups received 10 and 100 nM MitoQ compared to control group. Therefore, MitoQ is able to preserve quality parameters and fertility potential of post-thawed spermatozoa in rooster and it could be an effective additive for supplementation of rooster's semen cryopreservation medium during reproductive programs.

Alipour-Jenaghard P ，Daghigh-Kia H ，Masoudi R 《-》

Preservation of rooster post-thawed sperm epigenetic modifications, fertility potential and other quality parameters in different extenders using reduced glutathione.

Although rooster semen cryopreservation is an efficient procedure to spread qualified semen samples for reproductive goals, some post-thawed qualified semen samples resulted in poor fertility rate that could be related to epigenetic modifications during the cryopreservation process. This research was conducted to investigate the effect of reduced glutathione (GSH) in different cryopreservation extenders (Lake and Beltsville) on preservation of epigenetic modifications, fertility potential and other quality parameters of rooster sperm after thawing. Semen samples were collected and diluted in Lake and Beltsville extenders as follows: L-0: Lake without GSH, L-G: Lake with GSH, B-0: Beltsville without GSH, and B-G: Beltsville with GSH. After freeze-thawing process, sperm motility, membrane functionality, mitochondrial activity, acrosome integrity, viability, apoptosis status, lipid peroxidation, DNA fragmentation, ROS concentration, epigenetic modifications and fertility potential were evaluated. In results, the type of extender had no effect (P > 0.05) of post-thawed sperm quality. The treatments containing GSH presented higher (P ≤ 0.05) total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, viability, DNA methylation, fertility as well as lower (P ≤ 0.05) lipid peroxidation, apoptosis, DNA fragmentation and ROS concentration than other treatments. Extender supplementation with GSH had no effect (P > 0.05) on histone methylation, histone acetylation and hatching rate. In conclusion, supplementation of rooster sperm cryopreservation extender with GSH could be an effective strategy to preserve post-thawed sperm DNA methylation, fertility and other quality parameters during reproductive programs.

Masoudi R ，Hatami M ，Esmaeilkhanian S ，Zarei F ，Sharafi M ，Hatefi A ... -  《-》

Effects of freezing extender supplementation with mitochondria-targeted antioxidant Mito-TEMPO on frozen-thawed rooster semen quality and reproductive performance.

Rooster semen cryopreservation is a useful method to utilize semen samples for artificial insemination in commercial flocks, but with use of the freezing-thawing process there is a reduction in the quality and fertilization capacity of rooster spermatozoa post-thawing. The aim of the current study was to investigate the efficacy of the mitochondria-targeted antioxidant Mito-TEMPO on rooster sperm quality and fertilization capacity after conducting the freezing-thawing processes. Semen samples were diluted and there were five equal aliquots supplemented with 0, 0.5, 5, 50 and 500 μM Mito-TEMPO. Semen samples were subsequently cryopreserved in liquid nitrogen. After thawing, sperm motility, lipid peroxidation, membrane functionality, normal morphology, mitochondria active potential, acrosome integrity, viability, apoptotic-like changes, DNA fragmentation, hydrogen peroxide concentration and fertilizing capacity were evaluated. Supplementation of Lake medium with 5 and 50 μM Mito-TEMPO resulted in greater (P ≤ 0.05) total sperm motility, progressive motility, average path velocity, membrane functionality, mitochondria active potential, acrosome integrity and viability compared with semen of the other groups. Lipid peroxidation, late apoptotic-like changes, DNA fragmentation and hydrogen peroxide content, however, were less (P ≤0.05) in semen samples supplemented with 5 and 50 μM Mito-TEMPO compared to other groups. Furthermore, fertility percentages were greater when there was supplementation with 5 and 50 μM Mito-TEMPO compared to the control group. Mitochondria-targeted antioxidant Mito-TEMPO could be included in semen extender before cryopreservation to improve quality and fertilization capacity of rooster semen after thawing of cryopreserved samples.

Masoudi R ，Asadzadeh N ，Sharafi M 《-》

Mitochondria-targeted antioxidant "MitoQ" improves rooster's cooled sperm quality indicators and reproductive performance.

The cell membrane of rooster sperm is sensitive to cold due to the high content of polyunsaturated fatty acids, which are very susceptible to lipid peroxidation. The present study was conducted to determine the effect of different concentrations of the mitochondrial-targeting antioxidant "MitoQ" on sperm quality and fertility potential of chilled semen in roosters. Semen samples were collected from 10 roosters, diluted in Lake extender, assigned into 5 groups according to MitoQ concentrations (0, 1, 10, 100 and 1000 nM MitoQ) and stored at 5 °C up to 48 h. Motility, mitochondrial activity, viability, membrane integrity, and lipid peroxidation were assessed at 0, 24, and 48 h of cold storage periods. In addition, the fertility potential was assessed using 24 h-cooled semen samples. Our results showed that extender supplementation with MitoQ had no effect (P > 0.05) on chilled semen samples quality parameters at time 0, while at times 24 and 48 h storage, samples contained 100 nM MitoQ presented higher (P ≤ 0.05) total motility, progressive motility, viability and membrane integrity compared to the other groups. In addition, semen samples containing 10 and 100 nM MitoQ showed higher (P ≤ 0.05) mitochondrial activity and lower (P ≤ 0.05) lipid peroxidation than other groups at 24 and 48 h storage. Fertility rate was higher (P ≤ 0.05) when the hens were artificially inseminated with 24 h-chilled semen samples containing 100 nM MitoQ. In conclusion, supplementing Lake Extender with 100 nM MitoQ could be a helpful strategy to preserve chilled semen quality and fertility potential in the rooster.

Alipour-Jenaghard P ，Daghigh-Kia H ，Masoudi R ，Moghaddam G ，Qasemi-Panahi B ... -  《-》

The effects of MitoQ as a mitochondrial-targeted antioxidant in a plant-based extender on buck sperm quality parameters during cryopreservation.

Hatami M ，Masoudi R ，Hatefi A ，Alipour-Jenaghard P ，Esmaeili V ... -  《-》