Non-viable sperm in the ejaculate: Lethal escorts for contemporary viable sperm

Roca, Jordi; Parrilla, Inmaculada; Gil, M.A.; Cuello, C.; Martínez, E. A.; Rodriguez‐Martinez, Heriberto

doi:10.1016/j.anireprosci.2016.02.028

Cited by 36 publications

(39 citation statements)

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“…The authors suggest that differences in the biochemical components of the HEY, OEY, or LPFo could account for the marked variations in sperm cryo-survival. Another possible explanation for this phenomenon could be the high proportions of dead frozen-thawed spermatozoa in the lactose-OEY-G extender, which might be an additional source of production of reactive oxygen species (ROS) resulting in compromised sperm viability and DNA integrity (Roca et al, 2016). Similar assumptions have been suggested for the wide variations in cryo-induced sperm DNA damage for boar semen frozen in various avian egg yolk-based extenders (Strzeżek et al, 2014;Fraser et al, 2016).…”

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confidence: 76%

Sperm DNA damage in relation to lipid peroxidation following freezing-thawing of boar semen

et al. 2017

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This study investigated the relationships between lipid peroxidation (LPO) and sperm DNA damage following freezing-thawing of boar semen in different extenders. The comet assay was used to measure the extent of sperm DNA damage in a cryoprotectant-free extender or in cryoprotectant-based extenders after single and repeated freezing and thawing. As well as an analysis of sperm motion characteristics, mitochondrial function, membrane integrity, and lipid peroxidation (LPO) were assessed simultaneously with the measurements of sperm DNA damage. Consistent positive significant correlations were found between sperm DNA damage and LPO after freezing-thawing. Comet assay measurements showed that cryo-induced sperm DNA damage was more marked in the cryoprotectant-free extender, irrespective of freezing cycle. The frequency of sperm cells with damaged DNA increased with repeated freezing and thawing in the cryoprotectant-based extenders. Except for sperm DNA damage, there were no consistent associations between post-thaw sperm LPO and sperm quality characteristics. It could be suggested that the increased LPO of membrane phospholipids is associated with higher susceptibility of boar spermatozoa to cryoinduced DNA damage. ______________________________________________________________________________________ Keywords: Comet assay measurements, cryopreservation, extenders, spermatozoa # Corresponding author: fraser@uwm.edu.pl Evidence has shown that reduced fertilizing ability of frozen-thawed spermatozoa is because of compromised viability, DNA integrity, and increased peroxidative damage (Thompson et al., 2009;Fraser et al., 2010;Hamilton et al., 2016;Fraser et al., 2016). Intact sperm DNA integrity is essential for fertilization and embryo development (Evenson, 2016). Accumulating evidence has shown that sperm DNA damage is a useful marker for post-thaw semen quality (Fraser et al., 2010;Gosálvez et al., 2011;Gürler et al., 2016). Cryo-induced oxidative stress, associated with excess reactive oxygen species (ROS), has a negative effect on sperm nuclear DNA integrity (Peris et al., 2007; Thompson et al., 2009;Gürler et al., 2016). Moreover, lipid peroxidation (LPO) cascade is initiated by oxidative challenge on the membrane phospholipids of spermatozoa, which significantly compromises their function (Neild et al., 2005;Whitaker et al., 2012;Hamilton et al., 2016). Studies on the relationships between DNA damage and LPO of frozen-thawed boar spermatozoa are limited. This study therefore investigated the relationships between sperm DNA damage and LPO following freezing-thawing of boar semen in a cryoprotectant-free extender or in cryoprotectantbased extenders supplemented with whole hen egg yolk (HEY), ostrich egg yolk (OEY) and lipoprotein fractions isolated from OEY (LPFo). Repeated freezing and thawing of semen samples was performed to assess its effect on post-thaw sperm DNA integrity.Sperm-rich fractions (SRFs) were collected from five Polish Large White boars (average age 18 months), using the gloved-hand tech...

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confidence: 76%

Sperm DNA damage in relation to lipid peroxidation following freezing-thawing of boar semen

et al. 2017

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“…Martin-Muñoz et al [33] demonstrated assessing these markers to determine the differential effect of ROS during cryopreservation on stallion sperm and the importance of ROS homeostasis in the selection of ejaculates for freezing. The elimination of non-viable sperm from a sample could be important in ROS homeostasis since the dead sperm are known to have a negative effect on the viable subpopulation [9]. In addition, Roca et al [34] showed that high proportions of dead sperm in raw semen samples before and during freezing induce significantly increased ROS generation and nuclear DNA fragmentation in frozen-thawed sperm.…”

Section: Discussionmentioning

confidence: 99%

“…One of the most important objectives of conservationist projects is to store as many high-quality samples as possible. Sperm selection optimizes the use of available space, allowing us to select useful cells (viable and motile sperm) and discard non-viable, apoptotic and non- motile sperm that impair the quality of the sample [9]. …”

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confidence: 99%

Improving sperm banking efficiency in endangered species through the use of a sperm selection method in brown bear (Ursus arctos) thawed sperm

et al. 2017

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BackgroundSperm selection methods such as Single Layer Centrifugation (SLC) have been demonstrated to be a useful tool to improve the quality of sperm samples and therefore to increase the efficiency of other artificial reproductive techniques in several species. This procedure could help to improve the quality of genetic resource banks, which is essential for endangered species. In contrast, these sperm selection methods are optimized and focused on farm animals, where the recovery task is not as important as in endangered species because of their higher sperm availability. The aim of this study was to evaluate two centrifugation methods (300 x g/20 min and 600 x g/10 min) and three concentrations of SLC media (Androcoll-Bear −80, 65 and 50%) to optimise the procedure in order to recover as many sperm with the highest quality as possible. Sperm morphology could be important in the hydrodynamic relationship between the cell and centrifugation medium and thus the effect of sperm head morphometry on sperm yield and its hydrodynamic relationship were studied.ResultsThe samples selected with Androcoll-Bear 65% showed a very good yield (53.1 ± 2.9) although the yield from Androcoll-Bear 80% was lower (19.3 ± 3.3). The latter showed higher values of motility than the control immediately after post-thawing selection. However, both concentrations of colloid (65 and 80%) showed higher values of viable sperm and viable sperm with intact acrosome than the control. After an incubation of 2 h at 37 °C, the samples from Androcoll-Bear 80% had higher kinematics and proportion of viable sperm with intact acrosome. In the morphometric analysis, the sperm selected by the Androcoll-Bear 80% showed a head with a bigger area which was more elongated than the sperm from other treatments.ConclusionsWe conclude that sperm selection with Androcoll-Bear at either 65% or 80% is a suitable technique that allows a sperm population with better quality than the initial sample to be obtained. We recommend the use of Androcoll-Bear 65% since the yield is better than Androcoll-Bear 80%. Our findings pave the way for further research on application of sperm selection techniques to sperm banking in the brown bear.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-017-1124-2) contains supplementary material, which is available to authorized users.

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“…Because the ejaculate contains a heterogeneous suspension of spermatozoa, sperm sub-populations of different quality and functionality are present (8). D'Amours et al (9) suggested that qualitative and/or quantitative differences in protein expression among these sperm subpopulations probably resulted in an unbalanced interaction between spermatozoa and the changing surrounding environment during ejaculation.…”

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The Proteome of Pig Spermatozoa Is Remodeled During Ejaculation

Pérez-Patiño

Parrilla

et al. 2019

Molecular & Cellular Proteomics

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The present study identified and quantified using iTRAQ-based LC-MS/MS, 1,723 and 1,602 proteins, respectively, in mature boar spermatozoa retrieved from cauda epididymis and three distinct fractions of the ejaculate. A total of 974 of the identified and 960 of the quantified proteins belonged to the Sus scrofa taxonomy. An ANOVA test revealed that 32 Sus scrofa proteins implicated in sperm function showed quantitative differences among sperm sources. The findings demonstrated that the proteome of boar spermatozoa is remodeled during ejaculation. Graphical Abstract Highlights• Proteome of mature boar spermatozoa from cauda epididymal and ejaculated sources were analyzed by iTRAQ-based LC-MS/MS.• 1,723 sperm proteins identified (974 of Sus scrofa taxonomy).• 1,602 sperm proteins quantified (960 of Sus scrofa taxonomy).• 32 Sus scrofa sperm proteins were differentially expressed among sperm sources.• The proteome of boar spermatozoa is remodelled during ejaculation. Proteins are essential for sperm function, including their fertilizing capacity. Pig spermatozoa, emitted in well-defined ejaculate fractions, vary in their functionality, which could be related to different sperm protein composition. This study aimed (i) to update the porcine sperm proteome and (ii) to identify proteins differentially expressed in mature spermatozoa from cauda epididymis and those delivered in separate ejaculate fractions. Ejaculates from nine mature and fertile boars were manually collected in three separate portions: the first 10 ml of the sperm-rich ejaculate fraction (SRF), the rest of the SRF and the post-SRF. The contents of cauda epididymides of the boars were collected post-mortem by retrograde duct perfusion, generating four different semen sources for each boar. Following centrifugation, the resulting pellets of each semen source were initially pooled and later split to generate two technical replicates per source. The resulting eight sperm samples (two per semen source) were subjected to iTRAQ-based 2D-LC-MS/MS for protein identification and quantification. A total of 1,723 proteins were identified (974 of Sus scrofa taxonomy) and 1,602 of them were also quantified (960 of Sus scrofa taxonomy).After an ANOVA test, 32 Sus scrofa proteins showed quantitative differences (p < 0.01) among semen sources, which was particularly relevant for sperm functionality in the post-SRF. The present study showed that the proteome of boar spermatozoa is remodeled during ejaculation involving proteins clearly implicated in sperm function. The findings provide valuable groundwork for further studies focused on identifying protein biomarkers of sperm fertility.

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Non-viable sperm in the ejaculate: Lethal escorts for contemporary viable sperm

Cited by 36 publications

References 73 publications

Sperm DNA damage in relation to lipid peroxidation following freezing-thawing of boar semen

Sperm DNA damage in relation to lipid peroxidation following freezing-thawing of boar semen

Improving sperm banking efficiency in endangered species through the use of a sperm selection method in brown bear (Ursus arctos) thawed sperm

The Proteome of Pig Spermatozoa Is Remodeled During Ejaculation

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