Effects of boar variability on comet-detected sperm-DNA damage following cryopreservation

2018 ◽  
Vol 58 (2) ◽  
pp. 252 ◽  
Author(s):  
L. Fraser ◽  
Ł. Zasiadczyk ◽  
C. S. Pareek
Keyword(s):  
Dna Damage ◽  
Membrane Integrity ◽  
Tail Length ◽  
Dna Integrity ◽  
Comet Tail ◽  
Sperm Dna Damage ◽  
Type Iv ◽  
Sperm Dna ◽  
Damage Type

Assessment of sperm-DNA integrity is a crucial issue in male fertility. In the present study, parameters derived from the image analysis of comets after single-cell gel electrophoresis were used to analyse the types of DNA damage of frozen–thawed boar spermatozoa. Semen, frozen in a cryoprotectant-free extender or in cryoprotectant-based extenders, was analysed for DNA fragmentation and with the following comet tail measures: percentage DNA in comet tail, comet tail length and olive tail moment. The percentages of sperm DNA damage in the comet tails were classified as Type 0 (no DNA damage), Type I (very low DNA damage), Type II (light DNA damage), Type III (medium DNA damage) and Type IV (heavy DNA damage). Sperm motility characteristics and membrane integrity were assessed in the pre-freeze and frozen–thawed semen samples. Assessment of sperm DNA fragmentation and comet tail measures showed marked inter-boar variability following cryopreservation. However, consistent differences among the boars, with respect to cryo-induced sperm DNA damage, were detected by the comet tail length and olive tail moment. Besides Type IV, all types of DNA damage were detected in the cryoprotectant-based extenders. It was found that the frequency of Type II and Type III of DNA damage of frozen–thawed spermatozoa was significantly greater in the cryoprotectant-based and cryoprotectant-free extenders respectively. Deterioration in the quality of the sperm DNA integrity was concomitant with a marked decline in sperm motility characteristics, reduced plasma membrane integrity and higher lipid peroxidation and aspartate aminotransferase activity after cryopreservation. It can be suggested that the comet-assay parameters, coupled with routine laboratory tests, are useful to improve the sperm evaluations of post-thaw quality of semen from individual boars and would offer more comprehensive information for a better understanding of the degree of cryo-induced sperm-DNA damage.

3 SELECTION OF UNFRAGMENTED-DNA SPERMATOZOA FROM HEAT STRESSED MICE BY FEMALE UTERINE TRACT AND ZONA PELUCIDA BINDING

2009 ◽  
Vol 21 (1) ◽  
pp. 102
Author(s):  
J. D. Hourcade ◽  
M. Perez-Crespo ◽  
B. Pintado ◽  
A. Gutiérrez-Adán
Keyword(s):  
Heat Treatment ◽  
Dna Damage ◽  
Sperm Motility ◽  
Tail Length ◽  
Dna Integrity ◽  
Sperm Selection ◽  
Epididymal Sperm ◽  
Cleavage Rate ◽  
Sperm Dna ◽  
Selection Of

Physiological bases of the sperm selection processes within the female reproductive tract before they meet and fertilize the oocyte are unknown. The aim of this work was to determine if one of the keys of spermatozoa selection could be DNA integrity. It has been reported that sperm DNA damage does not impair in vitro fertilization (IVF). However, it has been suggested that the zona pelucida (ZP) is able to select spermatozoa with unfragmented DNA (Liu and Baker 2007 Hum. Reprod. 22, 1597–1602). In this work, DNA damage of spermatozoa was artificially induced by scrotal heat treatment (HT) (42°C, 30 min). Twenty-one days after the HT, spermatozoa were recovered from the epididymis caudae of CD1 mice and from the uterine horns near the cervix (Uc), from the uterine horns near the oviducts (Uo), and from the oviducts (Ov) of CD1 females 1–2 h after mating with HT and control males. In each region we determined numbers of spermatozoa, individual motility and sperm DNA integrity by COMET assay (% DNA in tail, tail length, and COMET moment was calculated). Also, females naturally mated either with HT or control males were killed at Day 14 of pregnancy, and number of foetuses and resorptions was recorded. Additionally, IVF was performed with epididymal sperm from HT or control males, Two hours after IVF attached and un-attached spermatozoa to the ZP were recovered and samples were evaluated for sperm motility (CASA), sperm zona-binding, and sperm DNA fragmentation (COMET). Also cleavage rate of fertilized oocytes with sperm from HT or control males was analyzed. One-way ANOVA was used to compare the results form each group. Epididymal sperm count (12*106 and 4.4*106 for control and HT respectively), sperm motility (75 and 21% respectively) and testis weight (133.90 and 68.76 mg, respectively) were significantly reduced after heat treatment (P < 0.001). For the heat treatment, COMET values decreased significantly during the transit from Uc to Uo and from Uo to Ov (Tail DNA: 25.7, 23.5, and 14.4% respectively, P < 0.01; Tail length: 38.4, 29.4, and 11.2 pixels, P < 0.001; COMET Moment: 12.5, 8.5, and 2 respectively, P < 0.001). Heat treatment reduced numbers of foetuses (7 ± 0.5 v. 5 ± 0.49, control and HT group, respectively), but number of resorptions was not altered. Spermatozoa bound per ZP in IVF experiments (55 ± 7 and 13 ± 6, control and HT, respectively) and cleavage rate (61 ± 1 v. 15 ± 6, control and HT, respectively) were significantly reduced in the HT group. Two hours after IVF, spermatozoa attached to the ZP in HT group showed a significant decrease in COMET parameters as in tail length (59.46 ± 2.895 v. 34.66 ± 3.531), and in tail moment compared with unattached spermatozoa. Our results indicate that DNA integrity sperm selection mechanisms are present in both the female tract and the ZP. We suggest that genital tract and sperm-ZP binding process plays an important role in selection of sperm with normal chromatin DNA.

The effect of cholesterol loaded cyclodextrins on post-thawing quality of buffalo semen in relation to sperm DNA damage and ultrastructure

2017 ◽  
Vol 17 (1) ◽  
pp. 42-50 ◽  
Author(s):  
Mohamed Aboul Ezz ◽  
Abd Elmonem Montasser ◽  
Mamdouh Hussein ◽  
Ashraf Eldesouky ◽  
Magdy Badr ◽  
...  
Keyword(s):  
Dna Damage ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Buffalo Semen

miR-424/322 is downregulated in the semen of patients with severe DNA damage and may regulate sperm DNA damage

2016 ◽  
Vol 28 (10) ◽  
pp. 1598 ◽  
Author(s):  
Kai Zhao ◽  
Yaoping Chen ◽  
Ruifeng Yang ◽  
Yang Bai ◽  
Cuiling Li ◽  
...  
Keyword(s):  
Dna Damage ◽  
Human Sperm ◽  
Vital Role ◽  
Dna Integrity ◽  
Dna Double Strand Breaks ◽  
Human Seminal Plasma ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Murine Homologue

Sperm DNA integrity is an essential factor for accurate transmission of genetic information. Human sperm DNA damage is a common cause of male infertility but the exact mechanism remains poorly understood. Considering the vital role of microRNA (miRNA) in multiple pathophysiological processes, we hypothesised that testicular miRNA is involved in sperm DNA damage during spermatogenesis. Infertile patients with high sperm DNA fragment index (DFI; n = 94) were selected from 1090 infertile men and a total of 18 testis-specific seminal miRNAs previously identified from human seminal plasma were chosen and tested. miR-29c and miR-424 were downregulated in men with high DFI. The inhibition of these two miRNAs in mice confirmed the role of miR-424 (murine homologue miR-322) in sperm DNA damage during spermatogenesis; by contrast, miR-29c exhibited a negative result. Thus, miR-424/322 is involved in sperm DNA damage. Furthermore, the dysregulation of this miRNA can induce DNA double-strand breaks during spermatogenesis.

scholarly journals Interspecific ICSI for the Assessment of Sperm DNA Damage: Technology Report

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1250
Author(s):  
Jana Rychtarova ◽  
Alena Langerova ◽  
Helena Fulka ◽  
Pasqualino Loi ◽  
Michal Benc ◽  
...  
Keyword(s):  
Dna Damage ◽  
Damage Level ◽  
Sperm Dna Damage ◽  
Mammalian Sperm ◽  
Sperm Dna

Xenogenic mammalian sperm heads injected into mouse ovulated oocytes decondense and form pronuclei in which sperm DNA parameters can be evaluated. We suggest that this approach can be used for the assessment of sperm DNA damage level and the evaluation of how certain sperm treatments (freezing, lyophilization, etc.) influence the quality of spermatozoa.

scholarly journals Effect of Alcohol Consumption on the Sperm DNA Integrity: A Systematic Review

2017 ◽  
Vol 9 (13) ◽  
pp. 136
Author(s):  
Farah Hanan Fathihah Jaafar ◽  
Khairul Osman ◽  
Jaya Kumar ◽  
Siti Fatimah Ibrahim
Keyword(s):  
Dna Damage ◽  
Alcohol Consumption ◽  
Alcohol Withdrawal ◽  
Semen Parameters ◽  
Sperm Chromatin ◽  
Dna Integrity ◽  
Sperm Dna Damage ◽  
Sperm Dna Integrity ◽  
Sperm Dna ◽  
Keywords Alcohol

There is no solid conclusion on the conventional sperm parameters in association with alcohol consumption, evaluation of sperm DNA integrity thus become a more reliable parameter. Hereby, this literature search was performed to summarize alcohol consumption on the sperm DNA integrity. A computerized database search was done through MEDLINE via Ovid (since 1946 until August 2017) and Cochrane was used. The following set of keywords: ‘alcohol consumption OR alcohol intake OR alcohol diet OR drinking alcohol OR ethanol diet’ AND ‘sperm DNA OR sperm chromatin OR sperm genome OR sperm histone OR sperm protamine’ were utilised. 24 articles were retrieved where only five studies conform to the inclusion criteria All studies demonstrated a negative effect of alcohol consumption on sperm DNA integrity, regardless of various range of alcohol doses and duration of alcohol consumption. Out of five studies reviewed, four studies were using a different approach to measure the sperm DNA damage. Hereby, this review identified a need to use a single approach of DNA damage test by having various method of alcohol administration and/or vice versa so that the extension of sperm DNA damage to alcohol consumption will have a better conclusion. On the same note, a few studies have reported the reversibility on conventional semen parameters, none has been done on the sperm DNA damage upon alcohol withdrawal. Therefore, the role of alcohol withdrawal on the reversibility of sperm DNA damage needs to be as well investigated further.

scholarly journals Impact of sperm DNA damage and oocyte-repairing capacity on trout development

Reproduction ◽  
2016 ◽  
Vol 152 (1) ◽  
pp. 57-67 ◽  
Author(s):  
C Fernández-Díez ◽  
S González-Rojo ◽  
M Lombó ◽  
M P Herráez
Keyword(s):  
Dna Damage ◽  
Environmental Changes ◽  
Oocyte Quality ◽  
Dna Integrity ◽  
Repair Capacity ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
Apoptotic Activity ◽  
Repair Genes ◽  
Hatching Rates

Zygotic repair of paternal DNA is essential during embryo development. In spite of the interest devoted to sperm DNA damage, its combined effect with defect-repairing oocytes has not been analyzed. Modification of the breeding season is a common practice in aquaculture. This practice reduces developmental success and could affect the both factors: sperm DNA integrity and oocyte repair capacity. To evaluate the maternal role, we analyzed the progeny outcome after fertilizing in-season trout oocytes with untreated and with UV-irradiated sperm. We also analyzed the offspring obtained out of season with untreated sperm. The analysis of the number of lesions in 4 sperm nuclear genes revealed an increase of 1.22–11.18 lesions/10 kb in out-of-season sperm, similar to that obtained after sperm UV irradiation (400 µW/cm25 min). Gene expression showed in out-of-season oocytes the overexpression of repair genes (ogg1, ung, lig3, rad1) and downregulation of tp53, indicating an enhanced repairing activity and reduced capacity to arrest development upon damage. The analysis of the progeny in out-of-season embryos revealed a similar profile tolerant to DNA damage, leading to a much lower apoptotic activity at organogenesis, lower hatching rates and increased rate of malformations. The effects were milder in descendants from in-season-irradiated sperm, showing an enhanced repairing activity at epibolia. Results point out the importance of the repairing machinery provided by the oocyte and show how susceptible it is to environmental changes. Transcripts related to DNA damage signalization and repair could be used as markers of oocyte quality.

scholarly journals A cost for high levels of sperm competition in rodents: increased sperm DNA fragmentation

2016 ◽  
Vol 283 (1826) ◽  
pp. 20152708 ◽  
Author(s):  
Javier delBarco-Trillo ◽  
Olga García-Álvarez ◽  
Ana Josefa Soler ◽  
Maximiliano Tourmente ◽  
José Julián Garde ◽  
...  
Keyword(s):  
Dna Damage ◽  
Sperm Competition ◽  
Dna Fragmentation ◽  
Sperm Chromatin ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna Damage ◽  
Sperm Dna Integrity ◽  
Sperm Dna ◽  
Physical And Chemical

Sperm competition, a prevalent evolutionary process in which the spermatozoa of two or more males compete for the fertilization of the same ovum, leads to morphological and physiological adaptations, including increases in energetic metabolism that may serve to propel sperm faster but that may have negative effects on DNA integrity. Sperm DNA damage is associated with reduced rates of fertilization, embryo and fetal loss, offspring mortality, and mutations leading to genetic disease. We tested whether high levels of sperm competition affect sperm DNA integrity. We evaluated sperm DNA integrity in 18 species of rodents that differ in their levels of sperm competition using the sperm chromatin structure assay. DNA integrity was assessed upon sperm collection, in response to incubation under capacitating or non-capacitating conditions, and after exposure to physical and chemical stressors. Sperm DNA was very resistant to physical and chemical stressors, whereas incubation in non-capacitating and capacitating conditions resulted in only a small increase in sperm DNA damage. Importantly, levels of sperm competition were positively associated with sperm DNA fragmentation across rodent species. This is the first evidence showing that high levels of sperm competition lead to an important cost in the form of increased sperm DNA damage.

scholarly journals 141 Antioxidant Supplementation Alleviates DNA Damage in Boar Sperm Induced by Tropical Heat Stress

2018 ◽  
Vol 30 (1) ◽  
pp. 210 ◽  
Author(s):  
S. T. Peña ◽  
B. Gummow ◽  
A. J. Parker ◽  
D. B. B. P. Paris
Keyword(s):  
Dna Damage ◽  
Heat Stress ◽  
Dna Integrity ◽  
Antioxidant Supplementation ◽  
Sperm Dna Damage ◽  
Progressive Motility ◽  
Sperm Dna Integrity ◽  
Sperm Dna ◽  
Boar Sperm ◽  
The Tropics

Seasonal heat stress is known to significantly diminish reproductive performance in pigs, particularly in the tropics, costing the industry millions in annual losses. The boar’s reduced capacity to sweat and non-pendulous scrotum, combined with the widespread use of European breeds in the tropics, makes this species particularly vulnerable to heat stress. Although heat stress is traditionally considered a sow problem, recent mouse studies demonstrate that heat stress-induced sperm DNA damage can result in arrested development and loss of early embryos. Our study investigated the impact of tropical summer heat stress on the quality and DNA integrity of boar sperm, and trialled antioxidant supplementation to alleviate the problem. Data, expressed as mean ± SEM, were analysed by one-way repeated-measures ANOVA with pairwise Bonferroni tests. Motility of sperm obtained from Large White boars (n = 5) housed in the dry tropics of Townsville, North Queensland, Australia, was characterised by computer-assisted sperm analysis but did not differ between summer, winter, or spring (total motility: 71.3 ± 8.1 v. 90.2 ± 4.2 v. 70.8 ± 5.5%, respectively; P > 0.05; progressive motility: 35.4 ± 7.0 v. 46.6 ± 4.0 v. 41.7 ± 2.8%, respectively; P > 0.05). Sperm DNA integrity in 20,000 sperm/boar per season, evaluated using TUNEL and flow cytometry, revealed 16-fold more DNA-damaged sperm in summer than winter, and nearly 9-fold more than spring (16.1 ± 4.8 v. 1.0 ± 0.2 v. 1.9 ± 0.5%, respectively; P ≤ 0.05). However, boar feed supplemented with 100 g/boar per day of proprietary custom-made antioxidants during summer significantly reduced sperm DNA damage to 9.9 ± 4.5% and 7.2 ± 1.6% (P ≤ 0.05) after 42 and 84 days of treatment respectively. Total and progressive motility were not altered by the supplement. In summary, sperm DNA integrity is compromised in boars during summer, suggesting that boar factors may contribute to seasonal embryo loss in sows. Moreover, such damage appears undetectable using traditional measures of sperm motility. Antioxidant supplementation during summer appears to mitigate the negative impact of heat stress on sperm DNA integrity.

Effect of tubal explants and their secretions on bovine spermatozoa: modulation of ROS production and DNA damage

2012 ◽  
Vol 24 (6) ◽  
pp. 871 ◽  
Author(s):  
Patricia Navarrete Gómez ◽  
Juan G. Alvarez ◽  
Jennie Risopatrón ◽  
Fernando Romero ◽  
Raúl Sánchez
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Physiological Role ◽  
Conditioned Media ◽  
Ros Generation ◽  
Dna Integrity ◽  
Ros Production ◽  
Sperm Dna Damage ◽  
Sytox Green ◽  
Sperm Dna

Although low levels of reactive oxygen species (ROS) play a physiological role in maintaining sperm function, an increase in ROS generation above these levels may result in the induction of sperm membrane and DNA damage. The main objective of this study was to determine whether bovine oviducal explants (TU) and their conditioned media (CM) have a modulatory effect on the production of ROS, and consequently, on sperm DNA integrity. Thawed sperm were exposed to bovine TU and to CM obtained from the ampullar and isthmal regions after 4 and 12 h, and DNA damage and intracellular ROS production was assessed by TUNEL and DHE and SYTOX Green, respectively. Co-incubation of spermatozoa with oviducal explants from the ampullar region (TUa) for 4 h resulted in a statistically significant increase in the percentage of spermatozoa with DNA damage compared with controls (P = 0.0106), and this increase was positively correlated with ROS levels. Conversely, although the incubation of spermatozoa with explants and conditioned media from the isthmal region (TUi and CMi, respectively) for 12 h resulted in an increase of spermatozoa with DNA damage compared with controls (P < 0.0001), this increase was not correlated with ROS levels. In conclusion, significant oxidative stress may take place in the oviduct, particularly during short-term incubation, and this may be related to changes in the antioxidant factors present in the oviducal cells and secretions. A redox imbalance in pro-oxidants and antioxidants in the oviduct may lead to oxidative stress and sperm DNA damage.

scholarly journals Can the SCD test and terminal uridine nick-end labeling by flow cytometry technique (TUNEL/FCM) be used interchangeably to measure sperm DNA damage in routine laboratory practice?

2019 ◽  
Vol 29 (1) ◽  
Author(s):  
Cécile Grèze ◽  
Aline Guttmann ◽  
Hanae Pons-Rejraji ◽  
Marie-Paule Vasson ◽  
Jacqueline Lornage ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Damage ◽  
Gradient Centrifugation ◽  
Intraclass Correlation ◽  
Systematic Difference ◽  
Test Reliability ◽  
Sperm Chromatin ◽  
Dna Integrity ◽  
Sperm Dna Damage ◽  
Sperm Dna

Abstract Background Numerous tests have been proposed to evaluate sperm DNA integrity. To assess the sperm chromatin dispersion (SCD) test in an andrology laboratory, twenty-five men attending Clermont-Ferrand (France) University Hospital’s Center for Reproductive Medicine were recruited. Sperm DNA damage was measured in the same semen samples using the SCD test and the Terminal Uridine Nick-end Labeling by flow cytometry technique (TUNEL/FCM) after density gradient centrifugation. Results SCD test reliability between readings, readers or slides was clearly established with very high agreement between measurements (Intraclass correlation coefficient (ICC) at 0.97, 0.95 and 0.98 respectively). Despite very good agreement between the SCD test and TUNEL/FCM (ICC at 0.94), the SCD test tended to slightly but significantly underestimate DNA damage compared with TUNEL (p = 0.0127). This systematic difference between the two techniques was − 3.39 ± 1.45% (mean ± SE). Conclusions Andrology laboratories using the SCD test to measure sperm DNA damage need to know that it appears to give slightly underestimated measurements compared to TUNEL/FCM. However, this systematic underestimation is very small in amplitude. Both techniques give almost perfectly congruent results. Our study underlines the importance for each laboratory to validate its method to assess sperm DNA damage before implementing it in routine andrology lab practice.

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