Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders

Gadea, J.; Molla, Md. Rezwan; Sellés, E.; Marco, Ma.; García–Vázquez, Francisco Alberto; Gardón, Juan Carlos

doi:10.1016/j.cryobiol.2010.12.001

Cited by 225 publications

(263 citation statements)

References 55 publications

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“…The improvement in the qualities of frozen-thawed boar semen found in the present study, might be explained by L-cysteine being a precursor of intracellular glutathione [6], whose level is decreased during the cryopreservation process, and subsequently resulting in a reduction of fertilization potential of frozen boar spermatozoa [29]. In addition, L-cysteine has the ability to pass into cells rapidly, and to be transformed into taurine, which, combined with a fatty acid in the plasma membrane, is transformed to acyl-taurine, which can improve surfactant properties and osmoregulation of the sperm membrane [24,28].…”

Section: Discussionsupporting

confidence: 50%

Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen

Kaeoket¹,

Chanapiwat²,

Tummaruk³

et al. 2010

Asian J Androl

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Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. The aim of this study was to determine the optimal concentration of L-cysteine needed for cryopreservation of boar semen. Twelve boars provided semen of proven motility and morphology for this study. The semen was divided into four portions in which the lactose-egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various concentrations of L-cysteine to reach 0 mmol L −1 (group I, control), 5 mmol L −1 (group II), 10 mmol L −1 (group III) and 15 mmol L −1 (group IV). Semen suspensions were loaded in straws (0.5 mL) and placed in a controlled-rate freezer. After cryopreservation, frozen semen samples were thawed and investigated for progressive motility, viability using SYBR-14/EthD-1 staining and acrosome integrity using FITC-PNA/EthD-1 staining. There was a significantly higher (P < 0.01) percentage of progressive motility, viability and acrosomal integrity in two L-cysteine-supplemented groups (group II and group III) compared with the control. There was a biphasic effect of L-cysteine, with the highest percentage of progressive motility, viability and acrosomal integrity in group III. In conclusion, 5 or 10 mmol L −1 was the optimum concentration of L-cysteine to be added to the LEY extender for improving the quality of frozen-thawed boar semen.

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Section: Discussionsupporting

confidence: 50%

Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen

Kaeoket¹,

Chanapiwat²,

Tummaruk³

et al. 2010

Asian J Androl

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“…On the basis of our results, we can conclude that 2.5 mM glutathione is not sufficient to protect the sperm from oxidative damage. In experiments carried out by Gadea et al (2005) and Yeste et al (2014), this positive effect was observed with the addition of 1 mM and 2 mM of GSH to the freezing extender, while Gadea et al (2004) reported that the addition of 5 mM GSH to the freezing extender did not result in any improvement in standard semen parameters. In our study, the addition of 5 mM GSH to the freezing extender resulted in: an improvement in motion parameters of thawed spermatozoa, a higher number of viable sperm and sperm with high ΔΨm, and a decrease in percentage of sperm with apoptotic-like changes (YO-PRO-1 + /PI -and AnV + /PI -).…”

Section: Discussionmentioning

confidence: 87%

“…During the cryopreservation process a significant reduction in the glutathione content of the semen was observed (Gadea et al 2004). This can be prevented by using glutathione-supplemented extender for cryopreservation.…”

Section: Discussionmentioning

confidence: 99%

Apoptotic-like changes of boar spermatozoa in freezing media supplemented with different antioxidants

Trzcińska¹,

Bryła²

2015

Polish Journal of Veterinary Sciences

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This study evaluated the effect of supplementing the freezing extender with exogenous antioxidants on apoptotic-like changes in post-thaw boar spermatozoa. A total of 36 ejaculates were resuspended in standard lactose-egg yolk-glycerol extender supplemented with antioxidant to final concentrations of 0 (as control), 2.5mM GSH (group I), 5.0 mM GSH (group II), 150 IU/mL SOD (group III), 300 IU/mL SOD (group IV), 200 IU/mL CAT (group V), 400 IU/mL CAT (group VI), 150 IU/mL SOD+200 IU/mL CAT (group VII), 300 IU/mL SOD+400 IU/mL CAT (group VIII). Sperm motility and apoptotic-like changes were determined before and after freeze-thawing. The various markers of apoptotic-like changes were measured: plasma membrane permeability by YO-PRO-1/PI assay, phosphatidylserine (PS) translocation across the plasma membrane using fluorescein-labeled Annexin-V, mitochondrial transmembrane potential detected by JC-1, and DNA fragmentation evaluated by TUNEL assay. The highest percentage of progressive motile sperm was noticed in group II (PM% 64.2±15.4) compared with control (PM% 36.8±5.5). The supplementation of 400 IU/mL CAT (group VI) revealed significant (P<0.01) reduction of apoptotic-like changes (YO-PRO-1 + /PI -: 13.1±7.5%, AnV + /PI -: 9.9±4.1%) in frozen-thawed spermatozoa compared with extender supplemented with 200 IU/mL CAT (group V). Irrespective of the concentration used, SOD and CAT in combination (group VII and group VIII) significantly (P<0.01) improved post-thaw sperm survival compared with the control. Evaluation by TUNEL assay revealed that cryopreservation and thawing did not induce DNA fragmentation in boar spermatozoa.

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“…It could be related to the pro-oxidative and anti-oxidative capacity that the antioxidant substances present at a different concentration, thereby producing paradoxical effects. In our previous experiments with respect to antioxidant addition of reduced glutathione (GSH) to freezing and thawing media [29][30][31]48], we detected more significant activities when the antioxidant was added in the thawing, than in the freezing media. A plausible explanation for this question would be that cryodamage of antioxidant enzymes during the freezing process results in an overall decrease of enzy matic antioxidant defences in the sperm.…”

Section: Resultsmentioning

confidence: 94%

Effect of genistein supplementation of thawing medium on characteristics of frozen human spermatozoa

Martínez-Soto¹,

DiosHourcade²,

Gutiérrez‐Adán³

et al. 2010

Asian J Androl

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In this study, we evaluated the effects of genistein supplementation of the thawing extender on frozen-thawed human semen parameters. We analyzed the effect of supplementation on sperm motility, capacitation (membrane lipid disorder), reactive oxygen species (ROS) generation, chromatin condensation and DNA damage. Using this preliminary information, it maybe possible to improve the cryopreservation process and reduce the cellular damage. We have confirmed that the isoflavone genistein (10 mmol L -1 ) has antioxidant properties on the frozen-thawed spermatozoa. This results in a decreased ROS production that shows a slight improvement in the sperm motility, and decreases the membrane lipid disorder and DNA damage caused by cryopreservation. These results suggest an effect of genistein on sperm functionality that could be of interest for assisted reproduction treatments using frozenthawed human spermatozoa, but further studies will be necessary to confirm our findings and to evaluate the possible clinical applications.

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Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders

Cited by 225 publications

References 55 publications

Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen

Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen

Apoptotic-like changes of boar spermatozoa in freezing media supplemented with different antioxidants

Effect of genistein supplementation of thawing medium on characteristics of frozen human spermatozoa

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