Kurumun Atıf Alan Makalesi
Atıf Alan Yayın
The influence of trehalose, taurine, cysteamine and hyaluronan on ram semen. Microscopic and oxidative stress parameters after freeze-thawing process
Scopus
Toplam 341 atıf DOI
There is a lack of information regarding lipid peroxidation and antioxidant capacity in cryopreserved ram semen, and cryopreservation is associated with the production of reactive oxygen species (ROS) which lead to lipid peroxidation (LPO) of sperm membranes, resulting in a loss of motility, viability and fertility of sperm. The aim of this study was to determine the influence of certain additives and their different doses on standard semen parameters, lipid peroxidation and antioxidant activities after the cryopreservation/thawing of ram semen. Ejaculates collected from four Akkaraman rams, a native breed of sheep, were evaluated and pooled at 33 °C. Semen samples which were diluted with a Tris-based extender containing additives including trehalose (50, 100 mM), taurine (25, 50 mM), cysteamine (5, 10 mM), and hyaluronan (0.5, 1 mg/ml), and an extender containing no additives (control) were cooled to 5 °C and frozen in 0.25 ml French straws, being stored in liquid nitrogen. Frozen straws were thawed individually at 37 °C for 20 s in a water bath for evaluation. The use of a Tris-based extender supplemented with 50 mM trehalose, 25 mM taurine, and 5 and 10 mM cysteamine led to higher percentages of post-thaw motility, in comparison to the control group (P < 0.01). No significant differences were observed in the percentages of acrosome and total abnormalities, and the hypoosmotic swelling test upon the supplementation of the freezing extender with antioxidants after the thawing of semen. In biochemical assays, the addition of antioxidants did not cause significant differences in levels of malondialdehyde (MDA), glutathione (GSH), and glutathione peroxidase (GSH-Px), after thawing, when compared to groups with no additives. In this study, catalase (CAT) activities were higher in the group that was applied 25 mM taurine as an antioxidant, than in all of the other groups (P < 0.001). Compared to the controls, antioxidant treatment with 100 mM trehalose, 50 mM taurine, 5 mM cysteamine and 0.5 mg/ml hyaluronan, significantly elevated vitamin E (vit E) levels in samples (P < 0.001).
Atıf Kaynağı
Atıf Yapan Yayın
The synergistic effect of trehalose and low concentrations of cryoprotectants can improve post-thaw ram sperm parameters
Scopus
Havuzumuzda 49 atıf almış
The objective of this study was to compare the effects of different concentrations of two different cryoprotectants (glycerol, G and ethylene glycol, EG) and trehalose (T), added to the semen extender, on post-thaw ram sperm parameters. Ejaculates, collected from 6 Merino rams, were pooled and evaluated at 37 °C. The pooled samples were divided into six equal aliquots, and diluted in Tris-based extenders containing 5% G, 3% G + 60 Mm T, 1.5% G + 100 Mm T, 5% EG, 3% EG + 60 mM T, and 1.5% EG + 100 Mm T. Subsequently, the samples were cooled to 5 °C, frozen in 0.25-ml French straws, and stored in liquid nitrogen (LN2). Frozen samples were thawed individually, at 37 °C for 25 s in a water bath, for evaluation. Sperm motility was assessed using a phase-contrast microscope with a warm stage maintained at 37 °C. Acrosome integrity (FITC/PNA-PI), sperm viability (SYBR-14/PI), mitochondrial activity (JC-1/PI), DNA damage (COMET assay) and DNA fragmentation (TUNEL test) were determined. The group of samples diluted in an extender containing 5% of glycerol (Group 5% G) displayed higher percentages of subjective motility, viability and mitochondrial activity of sperm, compared to the other groups (P < 0.05). On the other hand, Group 3% G + 60 mM T yielded the second-best results for subjective motility, viability and mitochondrial activity of sperm, when compared to the other groups. The post-thaw sperm parameters of Group 3% G + 60 Mm T did not show any statistically significant difference from those of Group 5% G. There were no statistically significant differences between the groups for acrosome integrity (P > 0.05). The results of the COMET assay showed that the use of low concentrations of cryoprotectants in combination with trehalose decreased sperm DNA damage. Accordingly, Group 1.5% G + 100 mM T and Group 3% EG + 60 mM T benefited from a significantly stronger cryoprotective effect on DNA integrity, in comparison to Group 5% G (P < 0.05). According to the results of the TUNEL test, the combined use of low concentrations of cryoprotectants with trehalose decreased sperm DNA damage, when compared to the use of 5% glycerol, but this difference was statistically insignificant (P > 0.05). In conclusion, G and EG concentrations can be reduced by adding various amounts of T (60 mM, 100 mM) to the semen extender. The addition of 5% of glycerol and 3% G + 60 mM T to the semen extender did not yield statistically different post-thaw sperm parameters, when compared for protection against cryoinjury. Post-thaw sperm parameters can be improved by the supplementation of the semen extender with 3% G + 60 mM T. Thus, we recommend the use of freezing extenders containing low cryoprotectant concentrations (3% G) combined with trehalose to avoid the high level of toxic and osmotic damage caused by 5% G.
Atıf Yapan Makale Bilgileri
Kurumlar (5)
Aksaray Üniversitesi
Aksaray, Turkey
Ankara Yildirim Beyazit University
Ankara, Turkey
Pamukkale Üniversitesi
Denizli, Turkey
Selçuk Üniversitesi
Selçuklu, Turkey
Tabriz University of Medical Sciences
Tabriz, Iran