scholarly journals Sperm FISH and chromatin integrity in spermatozoa from a t(6;10;11) carrier

Reproduction ◽  
2014 ◽  
Vol 147 (5) ◽  
pp. 659-670 ◽  
Author(s):  
Marta Olszewska ◽  
Nataliya Huleyuk ◽  
Monika Fraczek ◽  
Danuta Zastavna ◽  
Ewa Wiland ◽  
...  
Keyword(s):  
Blue Staining ◽  
Sperm Chromatin ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Sperm Aneuploidy ◽  
Sperm Dna ◽  
Sperm Fish ◽  
Chromatin Integrity ◽  
Meiotic Segregation Pattern ◽  
High Level

Complex chromosome rearrangements (CCRs) are structurally balanced or unbalanced aberrations involving more than two breakpoints on two or more chromosomes. CCRs can be a potential reason for genomic imbalance in gametes, which leads to a drastic reduction in fertility. In this study, the meiotic segregation pattern, aneuploidy of seven chromosomes uninvolved in the CCR and chromatin integrity were analysed in the ejaculated spermatozoa of a 46,XY,t(6;10;11)(q25.1;q24.3;q23.1)mat carrier with asthenozoospermia and a lack of conception. The frequency of genetically unbalanced spermatozoa was 78.8% with a prevalence of 4:2 segregants of 38.2%, while the prevalence of the adjacent 3:3 mode was 35.3%. Analysis of the aneuploidy of chromosomes 13, 15, 18, 21, 22, X and Y revealed an approximately fivefold increased level in comparison with that of the control group, indicating the presence of an interchromosomal effect. Sperm chromatin integrity status was evaluated using chromomycin A3 and aniline blue staining (deprotamination), acridine orange test and TUNEL assay (sperm DNA fragmentation). No differences were found when comparisons were made with a control group. We suggest that the accumulation of genetically unbalanced spermatozoa, significantly increased sperm aneuploidy level and decreased sperm motility (20%, progressive) were not responsible for the observed lack of reproductive success in the analysed infertile t(6;10;11) carrier. Interestingly, in the case described herein, a high level of sperm chromosomal imbalance appears not to be linked to sperm chromatin integrity status.

Sperm DNA fragmentation: causes and identification

Zygote ◽  
2019 ◽  
Vol 28 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Thais Rose dos Santos Hamilton ◽  
Mayra Elena Ortiz D’Ávila Assumpção
Keyword(s):  
Dna Fragmentation ◽  
Semen Analysis ◽  
Sperm Chromatin ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Functional Capability ◽  
Sperm Dna Integrity ◽  
Sperm Dna ◽  
Fertility Disorders ◽  
Chromatin Integrity

SummarySperm DNA fragmentation is referred to as one of the main causes of male infertility. Failures in the protamination process, apoptosis and action of reactive oxygen species (ROS) are considered the most important causes of DNA fragmentation. Action of ROS or changes in sperm protamination would increase the susceptibility of sperm DNA to fragmentation. Routine semen analysis is unable to estimate sperm chromatin damage. Sperm DNA integrity influences sperm functional capability, therefore tests that measure sperm DNA fragmentation are important to assess fertility disorders. Actually, there is a considerable number of methods for assessing sperm DNA fragmentation and chromatin integrity, sperm chromatin stability assay (SCSA modified), sperm chromatin dispersion (SCD), comet assay, transferase dUTP nick end labelling (TUNEL); and protamine evaluation in sperm chromatin assay, such as toluidine blue, CMA3, protamine expression and evaluation of cysteine radicals. This review aims to describe the main causes of sperm DNA fragmentation and the tests commonly used to evaluate sperm DNA fragmentation.

scholarly journals Effect of superoxide dismutase supplementation on sperm DNA fragmentation

2017 ◽  
Vol 89 (3) ◽  
pp. 212 ◽  
Author(s):  
Luciano Negri ◽  
Renzo Benaglia ◽  
Emanuela Monti ◽  
Emanuela Morenghi ◽  
Alessandro Pizzocaro ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Semen Parameters ◽  
Sperm Chromatin ◽  
Control Group ◽  
Sperm Dna Fragmentation ◽  
Antioxidant Supplementation ◽  
Sperm Dna ◽  
Dispersion Test ◽  
Antioxidant Supplements ◽  
Active Substances

Background: antioxidants supplementation improves sperm quality, but few trials have analyzed the effects on sperm DNA fragmentation (SDF). This study compares the effectiveness of SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol in reducing SDF with other antioxidants without SOD, hydroxytyrosol, and carnosol. Materials and methods: men with high SDF at baseline were selected in our clinical database. The patients taken into account had a 2-month control. SDF was measured by Sperm Chromatin Dispersion test (SCD). Untreated men were used as a control group. The remaining subjects received some oral antioxidant supplements (12 different combinations of both hydrophilic and lipophilic antioxidants), with some of them receiving nutritional support with a SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol. Results: 118 men were selected for a retrospective study. Mean age 39.3 ± 5.4 years. Fifteen had no treatment, 55 were treated with a SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol, and 48 took some antioxidant supplements for 2 months. Clinically, variations of at least 10% in baseline values of classic semen parameters and sperm DNA fragmentation were taken into consideration. Classic seminal parameters did not vary significantly in the three groups, with the exception of viability (p = 0.001). We assessed which of the active substances (no. 19) in different formulations were associated with variations in SDF. In the multivariable analysis of the 7 active substances that passed the univariable analysis, only the SOD molecule appeared to be linked to an improvement in SDF (< 0.0001). In detail, only one patient in the control group showed a spontaneous improvement in SDF (6%), compared to 16/48 (33%) of those taking various oral antioxidant supplements, and 31/55 (56%) of those taking a SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol. Conclusions: SOD-based antioxidant supplementation plus hydroxytyrosol and carnosol seems to provide a better chance of improving sperm DNA integrity than other classical antioxidant molecules.

scholarly journals Seminal Plasma Protein N-Glycan Peaks Are Potential Predictors of Semen Pathology and Sperm Chromatin Maturity in Men

Life ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 989
Author(s):  
Tihana Maric ◽  
Ana Katusic Bojanac ◽  
Ana Matijevic ◽  
Marcello Ceppi ◽  
Marco Bruzzone ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Plasma Protein ◽  
Seminal Plasma ◽  
Total Protein ◽  
Clinical Diagnostics ◽  
Semen Parameters ◽  
Blue Staining ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna

Background: Male infertility is increasingly becoming a health and demographic problem. While it may originate from congenital or acquired diseases, it can also result from environmental exposure. Hence, the complexity of involved molecular mechanisms often requires a multiparametric approach. This study aimed to associate semen parameters with sperm DNA fragmentation, chromatin maturity and seminal plasma protein N-glycosylation. Methods: The study was conducted with 166 participants, 20–55 y old, 82 normozoospermic and 84 with pathological diagnosis. Sperm was analyzed by Halosperm assay and aniline blue staining, while seminal plasma total protein N-glycans were analyzed by ultra-high-performance liquid chromatography. Results: Sperm DNA fragmentation was significantly increased in the pathological group and was inversely correlated with sperm motility and viability. Seminal plasma total protein N-glycans were chromatographically separated in 37 individual peaks. The pattern of seminal plasma N-glycan peaks (SPGP) showed that SPGP14 significantly differs between men with normal and pathological semen parameters (p < 0.001). The multivariate analysis showed that when sperm chromatin maturity increases by 10%, SPGP17 decreases by 14% while SPGP25 increases by 25%. Conclusion: DNA integrity and seminal plasma N-glycans are associated with pathological sperm parameters. Specific N-glycans are also associated with sperm chromatin maturity and have a potential in future fertility research and clinical diagnostics.

scholarly journals Sperm DNA Fragmentation, Determined Using the Sperm Chromatin Dispersion (SCD) Test, A Study in Republic of Kosovo Population

2018 ◽  
Vol 10 (3) ◽  
pp. 14
Author(s):  
Afrim Zeqiraj ◽  
Sheqibe Beadini ◽  
Nexhbedin Beadini ◽  
Hesat Aliu ◽  
Zafer Gashi ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Large Scale ◽  
Sperm Chromatin ◽  
Control Group ◽  
P Value ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Childbearing Age ◽  
Sperm Dna ◽  
The Republic

Infertility is a common condition affecting one in six couples of childbearing age. In approximately 40% of these cases, a male factor is involved. Sperm DNA integrity is essential for accurate transmission of genetic information. Materials and Methods: In this study 152 patients, 64 patients it is infertility group and 88 patients are fertile males. The ejaculate samples were taken in accordance with the patient to whom the reason for the analysis of the ejaculate sample was previously explained. All patients have been in the Dukagjini Region in the Republic of Kosovo. The samples were collected from 2016/18. Sperm Chromatin Dispersion (SCD) test, analysis in the ejaculate was performed at the Biolab Zafi, Laboratory in Peja, in the Republic of Kosovo. Statistical analysis: Data are reported as mean ± SD. The comparisons between groups were tested by student's t-test, ANOVA. A p-value less than 0.05% was considered statistically significant. Results: From our research studies, we have achieved significant (p &lt;.00001) scores among the working group and control group across all sperm parameters, and DNA fragmentation. Conclusion: In summary, we have demonstrated that there was a negative correlation between DNA fragmentation, sperm motility, and morphology in infertile males. We conclude that sperm DNA fragmentation appears to be a useful technique to predict outcome in couples undergoing IVF/ICSI. To evaluate whether DFI 23.94 ± 4.68% can be used to determine male infertility in our country by Sperm Chromatin Dispersion (SCD), it is necessary to carry out further large-scale research by other authors.

Spermatozoa of Sminthopsis murina (Mammalia: Metatheria) exhibit an unusually high degree of chromatin stability in the absence of disulphide bonding in protamine 1

2016 ◽  
Vol 28 (9) ◽  
pp. 1268 ◽  
Author(s):  
S. D. Johnston ◽  
C. López-Fernández ◽  
F. Arroyo ◽  
S. Fardell ◽  
R. Roy ◽  
...  
Keyword(s):  
Restriction Endonuclease ◽  
Dna Fragmentation ◽  
Reproductive Tract ◽  
Sperm Chromatin ◽  
Cysteine Residues ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna Damage ◽  
Sperm Dna ◽  
High Level ◽  
Protamine 1

Although all but a single genus (Planigale) of the metatheria so far examined contain no cysteine residues in protamine 1, we report a remarkable level of chromatin stability in the spermatozoa of the common dunnart, Sminthopsis murina. S. murina cauda epididymal spermatozoa and somatic epithelial cells were exposed to a combination of graded treatments to lyse sperm protein and induce sperm DNA damage via standard freeze–thaw protocols and post-thaw incubation at 37°C for 48 h, exposure to sodium nitroprusside (SNP) and the enzyme AluI restriction endonuclease. Sperm DNA fragmentation was assessed using the comet assay and sperm chromatin dispersal test. Although S. murina somatic cells showed DNA fragmentation following protein lysis and after treatment with all the protocols specifically designed to induce chromatin damage, sperm DNA fragmentation was only observed following moderate to severe proteolytic exposure and treatment with the restriction endonuclease; there was also an increase in the baseline halo of spermatozoa treated with an aggressive reducing agent, but no corresponding evidence of fragmented DNA, suggesting that cysteine residues may be functioning to conform tertiary and/or quaternary chromatin structure. Given that the protamine 1 of S. murina contains no cysteine, we suggest that the source of these residues is possibly the histone fraction of the chromatin and that the high level of stability is potentially related to prolonged sperm survival in the female’s reproductive tract.

Sperm DNA fragmentation measured by sperm chromatin dispersion impacts morphokinetic parameters, fertilization rate and blastocyst quality in ICSI treatments

Zygote ◽  
2021 ◽  
pp. 1-8
Author(s):  
Shikai Wang ◽  
Weihong Tan ◽  
Yueyue Huang ◽  
Xianbao Mao ◽  
Zhengda Li ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Embryo Quality ◽  
Fertilization Rate ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Blastocyst Formation ◽  
High Quality ◽  
Morphokinetic Parameters ◽  
Sperm Dna ◽  
Blastocyst Quality

Summary To determine the effects of sperm DNA fragmentation (SDF) on embryo morphokinetic parameters, cleavage patterns and embryo quality, this retrospective study analyzed 151 intracytoplasmic sperm injection (ICSI) cycles (1152 embryos collected) between November 2016 and June 2019. SDF was assessed using sperm chromatin dispersion. The cycles were divided into two groups based on the SDF rate: SDF < 15% (n = 114) and SDF ≥ 15% (n = 37). The embryo morphokinetic parameters, cleavage patterns, and embryo quality were compared between the two groups. The morphokinetic parameters tPNf, t2, t3, t4, t5, t6, and t8 were achieved significantly earlier in the SDF < 15% group compared with in the SDF ≥ 15% group. The fertilization and 2PN rates seemed to be significantly higher in the SDF < 15% group compared with in the SDF ≥ 15% group, while the abnormal cleavage rates were similar. However, a significantly higher rate of chaotic cleavage (CC) was observed in the SDF ≥ 15% group. The D3 high-quality embryo and available embryo rates were similar between the two groups. The blastocyst formation, high-quality blastocyst, and available blastocyst rates in the SDF < 15% group were significantly higher than those in the SDF ≥ 15% group. With an increase in SDF level, the chemical pregnancy, clinical pregnancy and implantation rates tended to decrease, while the miscarriage rate increased. This study demonstrated that SDF ≥ 15% reduces the fertilization rate of ICSI cycles and affects certain morphokinetic parameters. A higher SDF level can also induce a higher rate of CC, with subsequent decreases in the blastocyst formation rate and blastocyst quality.

scholarly journals 272 Proposing a combination of heritable fertility traits for bull selection

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 83-84
Author(s):  
Marina Fortes ◽  
Wei Liang Andre Tan ◽  
Laercio R Porto-Neto ◽  
Antonio Reverter ◽  
Gry B Boe-Hansen
Keyword(s):  
Dna Fragmentation ◽  
X Chromosome ◽  
Selective Breeding ◽  
Genetic Correlations ◽  
Research Data ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Sperm Dna ◽  
Fertility Traits ◽  
Protamine Deficiency

Abstract Traits such as sperm morphology and motility are routine in veterinarian evaluations of bull fertility. However, they rarely are included in livestock breeding programs, which typically use only scrotal circumference (SC) and some female traits for fertility selection. We studied 25 male fertility traits measured in two research populations of bulls (1,099 Brahman, and 1,719 Tropical Composite) and one commercial population (2,490 Santa Gertrude bulls). Measurements included standard semen evaluation (e.g. sperm motility and morphology) and SC. In the research data, we also measured sperm DNA fragmentation and sperm protamine deficiency for about 50% of the bulls. Using a mixture of genomic and pedigree analyses, we estimated heritabilities and genetic correlations for all traits, in each population. Our analyses suggest that bull fertility traits have a heritable component, which makes selective breeding possible. The phenotype variation in sperm DNA fragmentation and sperm protamine deficiency traits also have a heritable component (h2 ~ 0.05–0.22). These first estimates for heritability of sperm chromatin phenotypes require further studies, with larger datasets, to corroborate present results. In all three populations, we observed genetic correlations across traits that were favorable, but not high. For example, the percentage of normal sperm (PNS) from the sperm morphology evaluation was positively correlated with SC. In the research data, sperm DNA fragmentation was negatively correlated with PNS (r2 ~ 0.23–0.33), meaning that bulls with a higher PNS had less DNA fragmentation, being therefore more fertile according to both indicators. Given the favorable and yet not high genetic correlations between traits, it is possible to envision that sperm chromatin phenotypes might form a panel, together with PNS and SC, for a comprehensive bull fertility index. Selection indices that include fertility traits are being implemented in the dairy industry and could be recommended for beef cattle, too. An index that benefits from the favorable genetic correlations between traits that describe different aspects of bull fertility is a sensible approach to selective breeding. The clinical use of complementary indicators for male fertility is largely accepted, when deciding on bull fitness for the mating season. We propose extending this rationale to create a multi-trait index that captures genetic merit for bull fertility. In addition, we performed genome-wide association analyses in the research data and identified eight QTLs in the X chromosome. Correlations and shared SNP associations support the hypothesis that these phenotypes have the same underlying cause: abnormal spermatogenesis. In conclusion, it is possible to improve bull fertility through selective breeding, by measuring complementary fertility traits. Genomic selection for bull fertility might be more accurate if the X chromosome mutations that underlie the discovered QTL are included in the analyses. Polymorphisms associated with fertility in the bull accumulate in the X chromosome, as they do in humans and mice, thus suggesting specialization of this chromosome.

The assessment of sperm DNA fragmentation in the saltwater crocodile (Crocodylus porosus)

2017 ◽  
Vol 29 (3) ◽  
pp. 630 ◽  
Author(s):  
S. D. Johnston ◽  
C. López-Fernández ◽  
F. Arroyo ◽  
J. L. Fernández ◽  
J. Gosálvez
Keyword(s):  
Dna Damage ◽  
Dna Fragmentation ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Fragmented Dna ◽  
Sperm Dna ◽  
Saltwater Crocodile ◽  
Crocodylus Porosus ◽  
Dispersion Test ◽  
Sperm Nuclei

Herein we report a method of assessing DNA fragmentation in the saltwater crocodile using the sperm chromatin dispersion test (SCDt) after including frozen–thawed spermatozoa in a microgel (Halomax; Halotech DNA, Madrid, Spain). Following controlled protein depletion, which included a reducing agent, sperm nuclei with fragmented DNA showed a homogeneous and larger halo of chromatin dispersion with a corresponding reduced nucleoid core compared with sperm with non-fragmented DNA. The presence of DNA damage was confirmed directly by incorporation of modified nucleotides using in situ nick translation (ISNT) and indirectly by studying the correlation of the SCDt with the results of DNA damage visualisation using a two-tailed comet assay (r = 0.90; P = 0.037). Results of the SCDt immediately following thawing and after 5 h incubation at 37°C in order to induce a range of DNA damage revealed individual crocodile differences in both the baseline level of DNA damage and DNA longevity.

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