Antioxidants and their effect on the oxidative/nitrosative stress of frozen-thawed boar sperm

Pezo, Felipe; Yeste, Marc; Zambrano, Fabiola; Uribe, Pamela; Risopatrón, Jennie; Sánchez, Raúl

doi:10.1016/j.cryobiol.2020.11.007

Cited by 22 publications

(20 citation statements)

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“…Although, at first glance, the combination of these two results could seem contradictory, they may indicate that, while all post-thaw injury effects (targeting viability, motility and DNA damage) are originated from the same oxidative stress mechanism, the detrimental impact on some physiological parameters, such as viability and motility, occurs earlier. Our data support that the ROS generated during freezing and thawing processes (before our 0 h time point) [ 53 , 54 ] would oxidize lipids from mitochondrial and plasma membranes, leading to a rapid loss of mitochondrial membrane potential and plasma membrane integrity, which would ultimately abolish motility and induce cell death [ 10 , 43 ]. Hence, the constant reduction of total ROS observed throughout post-thawing incubation (i.e., between 0 and 4 h) could be the consequence of a rapid reaction of these highly reactive species to cell components and the leakage of ROS from non-viable sperm cells.…”

Section: Discussionsupporting

confidence: 76%

Determination of double- and single-stranded DNA breaks in bovine sperm is predictive of their fertilizing capacity

Ribas-Maynou

Delgado-Bermúdez

Mateo‐Otero

et al. 2022

J Animal Sci Biotechnol

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Background The analysis of chromatin integrity has become an important determinant of sperm quality. In frozen-thawed bovine sperm, neither the sequence of post-thaw injury events nor the dynamics of different types of sperm DNA breaks are well understood. The aim of the present work was to describe such sperm degradation aftermath focusing on DNA damage dynamics, and to assess if this parameter can predict pregnancy rates in cattle. Results A total of 75 cryopreserved ejaculates from 25 Holstein bulls were evaluated at two post-thawing periods (0-2 h and 2-4 h), analyzing global and double-stranded DNA damage through alkaline and neutral Comet assays, chromatin deprotamination and decondensation, sperm motility, viability, acrosomal status, and intracellular levels of total ROS, superoxides and calcium. Insemination of 59,605 females was conducted using sperm from the same bulls, thus obtaining the non-return to estrus rates after 90 d (NRR). Results showed an increased rate of double-stranded breaks in the first period (0-2 h: 1.29 ± 1.01%/h vs. 2-4 h: 0.13 ± 1.37%/h; P < 0.01), whereas the rate of sperm with moderate + high single-stranded breaks was higher in the second period (0-2 h: 3.52 ± 7.77 %/h vs. 2-4h: 21.06 ± 11.69 %/h; P < 0.0001). Regarding sperm physiology, viability decrease rate was different between the two periods (0-2 h: − 4.49 ± 1.79%/h vs. 2-4 h: − 2.50 ± 3.39%/h; P = 0.032), but the progressive motility decrease rate was constant throughout post-thawing incubation (0-2 h: − 4.70 ± 3.42%/h vs. 2-4 h: − 1.89 ± 2.97%/h; P > 0.05). Finally, whereas no correlations between bull fertility and any dynamic parameter were found, there were correlations between the NRR and the basal percentage of highly-damaged sperm assessed with the alkaline Comet ( Rs = − 0.563, P = 0.003), between NRR and basal progressive motility ( Rs = 0.511, P = 0.009), and between NRR and sperm with high ROS at 4 h post-thaw ( Rs = 0.564, P = 0.003). Conclusion The statistically significant correlations found between intracellular ROS, sperm viability, sperm motility, DNA damage and chromatin deprotamination suggested a sequence of events all driven by oxidative stress, where viability and motility would be affected first and sperm chromatin would be altered at a later stage, thus suggesting that bovine sperm should be used for fertilization within 2 h post-thaw. Fertility correlations supported that the assessment of global DNA damage through the Comet assay may help predict bull fertility. Supplementary Information The online version contains supplementary material available at 10.1186/s40104-022-00754-8.

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Section: Discussionsupporting

confidence: 76%

Determination of double- and single-stranded DNA breaks in bovine sperm is predictive of their fertilizing capacity

Ribas-Maynou

Delgado-Bermúdez

Mateo‐Otero

et al. 2022

J Animal Sci Biotechnol

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“…Uncontrolled (i.e. excessive) ROS generation can attack unsaturated fatty acids (UFAs) on the plasma membrane, alter mitochondrial membrane potential and induce protein sulfhydryl peroxidation, thereby altering sperm function and structure [13][14][15], which is regarded as an important cause of sperm damage during cryopreservation [16,17]. In addition, ROS could cause that the sperm be unable to fertilize the oocyte.…”

Section: Introductionmentioning

confidence: 99%

Freeze-thawing impairs the motility, plasma membrane integrity and mitochondria function of boar spermatozoa through generating excessive ROS

Zhang

Wang

et al. 2021

BMC Vet Res

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Background Cryopreservation is an efficient way to store spermatozoa and is closely associated with the quality of sperm after the freeze-thaw process. During freeze-thaw cycling, excessive reactive oxygen species (ROS) are produced, and the effects of ROS on boar sperm during cryopreservation have not been identified. Results In this study, we evaluated the quality of boar spermatozoa in different steps of cryopreservation (extension, cooling, and thawing for 30 min and 240 min) with or without boar-sperm antioxidant (N-acetylcysteine (NAC)). The ROS levels, sperm motility, plasma membrane integrity, mitochondrial activity, sperm chromatin structure, ATP content, and sperm apoptosis were assayed. After thawing, the ROS level and sperm apoptosis were significantly increased, and the sperm motility, plasma membrane integrity, mitochondrial activity, sperm chromatin structure, and ATP content were significantly impaired compared with those at the extension period and cooling period. Moreover, the addition of N-acetyl L-cysteine (NAC) reversed these changes. Conclusion The freeze-thawing of boar spermatozoa impaired their motility, plasma membrane, mitochondrial activity, sperm chromatin structure and apoptosis by producing excessive ROS. Thus, the downregulation of ROS level by antioxidants, especially the NAC, is important for manufacturing frozen pig sperm to increase reproductive cells and livestock propagation, as well as to improve the application of frozen semen in pigs worldwide.

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“…Boar spermatozoa are one of the most sensitive sperm types to cryopreservation. The resistance of spermatozoa to cryopreservation is highly dependent on their biophysical properties such as the shape, size, and plasma membrane composition, which is mainly linked to a lower cholesterol content and high content of polyunsaturated fatty acids [ 37 , 38 , 39 ].…”

Section: Discussionmentioning

confidence: 99%

Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose

Šimoník

Bubeníčková

Tůmová

et al. 2022

Animals

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The long-term storage of boar sperm presents an ongoing challenge, and the modification of the cryoprotective compounds in semen extenders is crucial for improving cryopreservation’s success rate. The aim of our study was to reduce the percentage of glycerol in the extender by elimination or substitution with biocompatible, non-toxic polysaccharides. For boar semen extender improvement, we tested a novel modification with the polysaccharides dextran and pentaisomaltose in combination with unique in silico predictive modeling. We targeted the analysis of in vitro qualitative sperm parameters such as motility, viability, mitochondrial activity, acrosome integrity, and DNA integrity. Non-penetrating polysaccharide-based cryoprotective agents interact with sperm surface proteins such as spermadhesins, which are recognized as fertility markers of boar sperm quality. The in silico docking study showed a moderate binding affinity of dextran and pentaisomaltose toward one specific spermadhesin known as AWN, which is located in the sperm plasma membrane. Pentaisomaltose formed a hydrophobic pocket for the AWN protein, and the higher energy of this protein–ligand complex compared with dextran was calculated. In addition, the root mean square deviation (RMSD) analysis for the molecular dynamics (MD) of both polysaccharides and AWN simulation suggests their interaction was highly stable. The in silico results were supported by in vitro experiments. In the experimental groups where glycerol was partially or entirely substituted, the use of pentaisomaltose resulted in improved sperm mitochondrial activity and DNA integrity after thawing when compared with dextran. In this paper, we demonstrate that pentaisomaltose, previously used for cryopreservation in hematopoietic stem cells, represents a promising compound for the elimination or reduction of glycerol in extenders for boar semen cryopreservation. This novel approach, using in silico computer prediction and in vitro testing, represents a promising technique to help identify new cryoprotectants for use in animal breeding or genetic resource programs.

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Antioxidants and their effect on the oxidative/nitrosative stress of frozen-thawed boar sperm

Cited by 22 publications

References 86 publications

Determination of double- and single-stranded DNA breaks in bovine sperm is predictive of their fertilizing capacity

Determination of double- and single-stranded DNA breaks in bovine sperm is predictive of their fertilizing capacity

Freeze-thawing impairs the motility, plasma membrane integrity and mitochondria function of boar spermatozoa through generating excessive ROS

Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose

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