The synergistic effect of trehalose and low concentrations of cryoprotectants can improve post-thaw ram sperm parameters
| dc.contributor.author | Öztürk, Ali Erdem | |
| dc.contributor.author | Bodu, Mustafa | |
| dc.contributor.author | Bucak, Mustafa Numan | |
| dc.contributor.author | Ağır, Vahit | |
| dc.contributor.author | Özcan, Ayşe | |
| dc.contributor.author | Keskin, Nazan | |
| dc.contributor.author | İli, Pınar | |
| dc.contributor.author | Topraggaleh, Tohid Rezaei | |
| dc.contributor.author | Sidal, Hümeyra | |
| dc.contributor.author | Başpınar, Nuri | |
| dc.contributor.author | Dursun, Şükrü | |
| dc.date.accessioned | 2020-04-16T07:03:37Z | |
| dc.date.available | 2020-04-16T07:03:37Z | |
| dc.date.issued | 2020 | |
| dc.department | Veteriner Fakültesi | |
| dc.description | Dursun, Şükrü ( Aksaray, Yazar ) | |
| dc.description.abstract | 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. | |
| dc.identifier.doi | 10.1016/j.cryobiol.2020.03.008 | |
| dc.identifier.endpage | - | en_US |
| dc.identifier.issn | 0011-2240 | |
| dc.identifier.issue | - | en_US |
| dc.identifier.pmid | 32259524 | |
| dc.identifier.scopusquality | Q1 | |
| dc.identifier.startpage | - | en_US |
| dc.identifier.uri | https:/dx.doi.org/10.1016/j.cryobiol.2020.03.008 | |
| dc.identifier.uri | https://hdl.handle.net/20.500.12451/7534 | |
| dc.identifier.volume | - | en_US |
| dc.identifier.wos | WOS:000556580400019 | |
| dc.identifier.wosquality | Q2 | |
| dc.indekslendigikaynak | Web of Science | |
| dc.indekslendigikaynak | Scopus | |
| dc.indekslendigikaynak | PubMed | |
| dc.language.iso | en | |
| dc.publisher | Academic Press Inc. | |
| dc.relation.ispartof | Cryobiology | |
| dc.relation.publicationcategory | Makale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı | |
| dc.rights | info:eu-repo/semantics/embargoedAccess | |
| dc.subject | Ethylene Glycol | |
| dc.subject | Glycerol | |
| dc.subject | Ram Semen | |
| dc.subject | Sperm Parameters | |
| dc.subject | Trehalose | |
| dc.title | The synergistic effect of trehalose and low concentrations of cryoprotectants can improve post-thaw ram sperm parameters | |
| dc.type | Article |
