Effects of reduced glutathione on the quality of rooster sperm during cryopreservation.

The purpose of this study was to investigate the beneficial effects of reduced glutathione (GSH) for cryopreservation of rooster semen. In experiment 1, semen samples were collected from 15 roosters and diluted in the Lake extender that contained various concentrations of GSH as follows: Lake without GSH (control, GSH 0), Lake containing 0.5 mM (GSH 0.5), 1 mM (GSH 1), 2 mM (GSH 2), 4 mM (GSH 4) and 8 mM (GSH 8) GSH. Viability, membrane functionality, morphology, mitochondrial activity, acrosome integrity, motion parameters, lipid peroxidation and DNA fragmentation were assessed after thawing. In experiment 2, reproductive performance of thawed semen was evaluated via artificial insemination. Supplemented extenders with 2 and 4 mM GSH presented higher (P ≤ 0.05) viability (59.4 ± 2.4% and 60.8 ± 2.4%), membrane functionality (62.3 ± 2.6% and 64.7 ± 2.6%), mitochondrial activity (49.4 ± 1.7% and 49.8 ± 1.7%), total motility (57.1 ± 1.9% and 58.8 ± 1.9%, respectively), progressive motility (28.9 ± 1.3% and 29.6 ± 1.3%), and lower lipid peroxidation (2.4 ± 0.09 nmol/ml and 2.3 ± 0.09 nmol/ml) compared to control group. Acrosome integrity was higher (P ≤ 0.05) in GSH 4 (91.4 ± 1.8%) compared to other groups. DNA fragmentation and MDA concentrations were higher (P ≤ 0.05) in GSH 8 (12 ± 1.2% and 3.4 ± 0.09 nmol/ml). In experiment 2, higher (P ≤ 0.05) fertility rate was observed in GSH 2 and GSH 4 (61.9% and 63.8%, respectively) compared to control (41.4%) group. In conclusion, supplementation of Lake extender with 2 and 4 mM GSH improves the cryo-survival and fertility potential of rooster sperm and it could be an applied method for improvement of reproductive goals.

Masoudi R ，Sharafi M ，Shahneh AZ ，Khodaei-Motlagh M ... -  《-》

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 ... -  《-》

Supplementation of extender with coenzyme Q10 improves the function and fertility potential of rooster spermatozoa after cryopreservation.

The effects of coenzyme Q10 (CoQ10) has not yet been assessed for cryopreservation of rooster semen. The aim of this study was to evaluate the effect of different concentrations of CoQ10 in Lake extender for cryopreservation of rooster semen. The viability and apoptosis status, DNA fragmentation, abnormal morphology, motion parameters, membrane functionality, mitochondrial activity, acrosome integrity, lipid peroxidation, and fertility potential were evaluated after the freeze-thaw process. Semen samples were collected from ten roosters, twice a week, and then diluted in extender contained different concentrations of CoQ10 as follows: Lake without CoQ10 (control, Q 0), Lake containing 1 μM (Q 1), 2 μM (Q 2), 5 μM (Q 5), and 10 μM (Q 10) CoQ10. Supplementation of Lake with 1 and 2 μM CoQ10 resulted in greater sperm viability, total motility, progressive motility, membrane functionality, mitochondrial activity, acrosome integrity, and fertility rate. Furthermore, the extent of lipid peroxidation in thawed spermatozoa treated with 1 and 2 μM CoQ10 was less than with the other groups. Different concentrations of CoQ10 had no effect on DNA fragmentation and sperm morphology. Results of the present study indicate that supplementation of Lake extender with 1 and 2 μM CoQ10 enhances the quality of rooster sperm after the freeze-thaw process.

Masoudi R ，Sharafi M ，Zare Shahneh A ，Kohram H ，Nejati-Amiri E ，Karimi H ，Khodaei-Motlagh M ，Shahverdi A ... -  《-》

Zinc oxide nanoparticles preserve the quality and fertility potential of rooster sperm during the cryopreservation process.

Karimi-Sabet MJ ，Khodaei-Motlagh M ，Masoudi R ，Sharafi M ... -  《-》

Supplementation of chilling storage medium with glutathione protects rooster sperm quality.

This study was aimed to evaluate the effect of addition of reduced glutathione (GSH) to the extender on the rooster's semen quality parameters and fertility potential. Semen samples were diluted with Lake extender contained 0, 0.5, 1, 2, 4 and 8 mM GSH. Then, were chilled to 5 °C and stored for a period of 48 h. Sperm motion characteristics, viability, membrane integrity, lipid peroxidation, mitochondrial activity and fertility potential were evaluated. At the initiation of the experiment (0 h), GSH did not affect sperm parameters, while 2-4 mM GSH improved (P ≤ 0.05) quality indicators during storage periods. Moreover, the samples treated with 2-4 mM GSH have had a lower lipid peroxidation compared to other groups (P ≤ 0.05). Artificial insemination using the semen samples, which had been stored in groups treated with 2-4 mM GSH for a period of 24 h, led to greater (P ≤ 0.05) fertilizing potential compared to the control group.

Masoudi R ，Sharafi M ，Pourazadi L ，Dadashpour Davachi N ，Asadzadeh N ，Esmaeilkhanian S ，Dirandeh E ... -  《-》