Effects of cryopreservation on human sperm deoxyribonucleic acid integrity

Zribi, Nassira; Chakroun, N.; Euch, Henda El; Gargouri, Jalel; Bahloul, Ali; Keskes, Leila

doi:10.1016/j.fertnstert.2008.09.038

Cited by 170 publications

(121 citation statements)

References 45 publications

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“…Thus, cryostorage is wellknown to be associated with the induction of DNA damage and, again, a majority of this damage seems to be oxidative. 58,59 Similarly, heat exposure will induce free radical generation by the spermatozoa as a consequence of the initiation of apoptotic cascade, and so will exposure to radiofrequency electromagnetic radiation 21 or hydrogen peroxide. 52 All of these conditions are associated with the creation of oxidative stress.…”

Section: Triggers Of Apoptosismentioning

confidence: 99%

Apoptosis and DNA damage in human spermatozoa

Aitken¹,

Koppers²

2010

Asian J Androl

286

179

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DNA damage is frequently encountered in spermatozoa of subfertile males and is correlated with a range of adverse clinical outcomes including impaired fertilization, disrupted preimplantation embryonic development, increased rates of miscarriage and an enhanced risk of disease in the progeny. The etiology of DNA fragmentation in human spermatozoa is closely correlated with the appearance of oxidative base adducts and evidence of impaired spermiogenesis. We hypothesize that oxidative stress impedes spermiogenesis, resulting in the generation of spermatozoa with poorly remodelled chromatin. These defective cells have a tendency to default to an apoptotic pathway associated with motility loss, caspase activation, phosphatidylserine exteriorization and the activation of free radical generation by the mitochondria. The latter induces lipid peroxidation and oxidative DNA damage, which then leads to DNA fragmentation and cell death. The physical architecture of spermatozoa prevents any nucleases activated as a result of this apoptotic process from gaining access to the nuclear DNA and inducing its fragmentation. It is for this reason that a majority of the DNA damage encountered in human spermatozoa seems to be oxidative. Given the important role that oxidative stress seems to have in the etiology of DNA damage, there should be an important role for antioxidants in the treatment of this condition. If oxidative DNA damage in spermatozoa is providing a sensitive readout of systemic oxidative stress, the implications of these findings could stretch beyond our immediate goal of trying to minimize DNA damage in spermatozoa as a prelude to assisted conception therapy.

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Section: Triggers Of Apoptosismentioning

confidence: 99%

Apoptosis and DNA damage in human spermatozoa

Aitken¹,

Koppers²

2010

Asian J Androl

286

179

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“…4,15,71,72 Recent studies have shown that DNA fragmentation can be primarily attributed to cryopreservation-induced ROS. [73][74][75] In contrast to superoxide anion, the more stable H 2 O 2 readily crosses the plasma membrane and is responsible for DNA oxidative damage. 76 Because DNA integrity is essential for achieving fertilisation and enabling embryo development in assisted reproductive technologies, the preservation of DNA during cryostorage is of extreme importance.…”

Section: Gangliosides and Spermatozoamentioning

confidence: 99%

Protective effects of exogenous gangliosides on ROS-induced changes in human spermatozoa

Gavella¹,

Lipovac²

2013

Asian J Androl

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This article summarizes the available evidence on the efficacy of gangliosides to reduce the degree of reactive oxygen species (ROS)-mediated damage. The antioxidative efficacy of exogenous gangliosides in protecting different cells encouraged us to examine their ability to protect human spermatozoa. Gangliosides are sialic acid-containing glycosphingolipids with strong amphiphilic character due to the bulky headgroup made of several sugar rings with sialic acid residues and the double-tailed hydrophobic lipid moiety. The amphiphilicity of gangliosides allows them to exist as micelles in aqueous media when they are present at a concentration above their critical micellar concentration. The protective effect of ganglioside micelles on spermatozoa is believed to stem from their ability to scavenge free radicals and prevent their damaging effects. In our study, we particularly focused our attention on the protective effect of ganglioside micelles on DNA in human spermatozoa exposed to cryopreservation. The results indicate that ganglioside micelles can modulate the hydrophobic properties of the sperm membrane to increase tolerance to DNA fragmentation, thus protecting the DNA from cryopreservation-induced damage. Further actions of ganglioside micelles, which were documented by biochemical and biophysical studies, included (i) the modulation of superoxide anion generation by increasing the diffusion barrier for membrane events responsible for signal translocation to the interior of the cell; (ii) the inhibition of iron-catalysed hydroxyl radical formation due to the iron chelation potential of gangliosides; and (iii) inhibition of hydrogen peroxide diffusion across the sperm membrane.

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“…However, these methods are presently criticized as affected by the possible spontaneous formation of 8-OHdG during the step of extraction/digestion of sperm DNA (Badouard et al 2008). Recently, emerging techniques to reveal 8-OHdG that employ antibodies (Kao et al 2008) or binding proteins , Zribi et al 2010 have been reported.…”

Section: Introductionmentioning

confidence: 99%

Development of a specific method to evaluate 8-hydroxy,2-deoxyguanosine in sperm nuclei: relationship with semen quality in a cohort of 94 subjects

Cambi¹,

Tamburrino²,

Marchiani³

et al. 2013

Reproduction

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Oxidative stress (OS) is involved in many disoders including male infertility. Human spermatozoa are very sensitive targets of reactive oxygen species (ROS) and most sperm functions are impaired in the case of OS. In addition unbalanced production of ROS is considered one of the most important causes of sperm DNA fragmentation, a semen trait of infertile men. The relationship between oxidative damage and semen quality is partially controversial, probably due to the different methods and/or targets used to reveal the OS. In this study, by fluorescence microscopy and flow cytometry, we compared two methods to reveal 8-hydroxy,2-deoxyguanosine (8-OHdG), the hallmark of oxidative DNA damage: an immunofluorescence method and the commercial OxyDNA kit. We found that although both methods localized the labelling in sperm nuclei they yielded different measures, and only with the immunofluorescence method was the labelling specific for sperm 8-OHdG. The immunofluorescence method, coupled to flow cytometry, was thus selected to analyse the 8-OHdG content in semen samples from 94 subfertile patients and to investigate the relationship with semen quality. We found that the percentages of spermatozoa with 8-OHdG (meanGS.D., 11.4G6.9%) were related to sperm count (Pearson's correlation coefficient (r)ZK0.27, PZ0.04 (ANOVA and student's t-test)), motility (progressive: rZK0.22, PZ0.04; non-progressive: rZ0.25, PZ0.01), and normal morphology (rZK0.27, PZ0.01).In conclusion, we demonstrate that immunofluorescence/flow cytometry is a reliable and specific method to detect 8-OHdG at single-cell level and show that oxidative damage only partially overlaps poor semen quality, suggesting that it could provide additional information on male fertility with respect to routine semen analysis.

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Effects of cryopreservation on human sperm deoxyribonucleic acid integrity

Cited by 170 publications

References 45 publications

Apoptosis and DNA damage in human spermatozoa

Apoptosis and DNA damage in human spermatozoa

Protective effects of exogenous gangliosides on ROS-induced changes in human spermatozoa

Development of a specific method to evaluate 8-hydroxy,2-deoxyguanosine in sperm nuclei: relationship with semen quality in a cohort of 94 subjects

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