The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100 μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1 h with 0, 20 and 100 µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (P<0.05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8 ± 1.5 and 69.1 ± 1.4 respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (P<0.05), while the percentage of spermatozoa with high ΔΨm increased (P<0.05). The results of Experiment 2 showed that 1 h-treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4 ± 5.8 and 83.7.1 ± 1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (P<0.05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (P<0.05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0 ± 11.4). In conclusion, the treatment with 100 µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100 µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation.
Effect of caspase inhibitor Z-VAD-FMK on bovine sperm cryotolerance / Pagano, Nunzia; Longobardi, Valentina; De Canditiis, Carolina; Zuchegna, Candida; Romano, Antonella; Kosior, Michal Andrzej; Pero, Maria Elena; Gasparrini, Bianca. - In: REPRODUCTION IN DOMESTIC ANIMALS. - ISSN 0936-6768. - (2020). [10.1111/rda.13648]
Effect of caspase inhibitor Z-VAD-FMK on bovine sperm cryotolerance
Pagano, Nunzia;Longobardi, Valentina;De Canditiis, Carolina;Zuchegna, Candida;Kosior, Michal Andrzej;Pero, Maria Elena
;Gasparrini, Bianca
2020
Abstract
The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100 μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1 h with 0, 20 and 100 µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (P<0.05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8 ± 1.5 and 69.1 ± 1.4 respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (P<0.05), while the percentage of spermatozoa with high ΔΨm increased (P<0.05). The results of Experiment 2 showed that 1 h-treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4 ± 5.8 and 83.7.1 ± 1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (P<0.05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (P<0.05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0 ± 11.4). In conclusion, the treatment with 100 µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100 µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation.File | Dimensione | Formato | |
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Reprod Domestic Animals - 2020 - Pagano - Effect of caspase inhibitor Z‐VAD‐FMK on bovine sperm cryotolerance.pdf
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