DNA methylation patterns vary in boar sperm cells with different levels of DNA fragmentation

Abstract Background Sperm DNA integrity is considered essential for successful transmission of the paternal genome, fertilization and normal embryo development. DNA fragmentation index (DFI, %) has become a key parameter in the swine artificial insemination industry to assess sperm DNA integrity. Recently, in some elite Norwegian Landrace boars (boars with excellent field fertility records), a higher level of sperm DFI has been observed. In order to obtain a better understanding of this, and to study the complexity of sperm DNA integrity, liquid preserved semen samples from elite boars with contrasting DFI levels were examined for protamine deficiency, thiol profile and disulphide bonds. Additionally, the DNA methylation profiles of the samples were determined by reduced representation bisulphite sequencing (RRBS). Results In this study, different traits related to sperm DNA integrity were investigated (n = 18 ejaculates). Upon liquid storage, the levels of total thiols and disulphide bonds decreased significantly, while the DFI and protamine deficiency level increased significantly. The RRBS results revealed similar global patterns of low methylation from semen samples with different levels of DFI (low, medium and high). Differential methylation analyses indicated that the number of differentially methylated cytosines (DMCs) increased in the low-high compared to the low-medium and the medium-high DFI groups. Annotating the DMCs with gene and CpG features revealed clear differences between DFI groups. In addition, the number of annotated transcription starting sites (TSS) and associated pathways in the low-high comparison was greater than the other two groups. Pathway analysis showed that genes (based on the closest TSS to DMCs) corresponding to low-high DFI comparison were associated with important processes such as membrane function, metabolic cascade and antioxidant defence system. Conclusion To our knowledge, this is the first study evaluating DNA methylation in boar sperm cells with different levels of DFI. The present study shows that sperm cells with varying levels of DNA fragmentation exhibit similar global methylation, but different site-specific DNA methylation signatures. Moreover, with increasing DNA fragmentation in spermatozoa, there is an increase in the number of potentially affected downstream genes and their respective regulatory pathways.

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Relationship of DNA integrity to HRG C633T SNP and ART outcome in infertile couples

Reproduction ◽

10.1530/rep-17-0058 ◽

2017 ◽

Vol 153 (6) ◽

pp. 865-876 ◽

Cited By ~ 1

Author(s):

Arumugam Kumaresan ◽

Anders Johannisson ◽

Sarah Nordqvist ◽

Karin Kårehed ◽

Helena Åkerud ◽

...

Keyword(s):

Dna Integrity ◽

Male Factor ◽

Seminal Parameters ◽

Sperm Dna Integrity ◽

Disulphide Bonds ◽

Sperm Dna ◽

Significant Difference ◽

Unknown Factor ◽

Art Outcome ◽

Protamine Deficiency

The status of sperm DNA fragmentation, protamine deficiency, free thiols and disulphide bonds in colloid-selected samples and its relationship to ART outcome orHRGC633T SNP is not known. The objective of this study was to determine these relationships in spermatozoa from men with male factor or unknown factor infertility (n = 118) undergoingin vitrofertilisation (IVF) or intracytoplasmic sperm injection (ICSI). Sperm DNA integrity was analysed by flow cytometry using three fluorescent probes (acridine orange, monobromobimane and chromomycin A3). Principal component analysis (PCA) was used to identify the parameters that most influenced fertility. The relationships of sperm DNA integrity with seminal parameters,HRGC633T SNP and ART outcome were established using ANOVA andt-test. Sperm concentration and yield after preparation accounted for 27% of the total variance; sperm DNA integrity (%DFI and disulphide bonds) accounted for 16% of the variance in men from infertile couples. Sperm %DFI was significantly higher (P < 0.05) in older men than in younger men. A significant difference (P < 0.01) was observed in %DFI between smokers and non-smokers. Sperm %DFI was significantly higher (P < 0.01) in male factor infertility compared to either female factor or unknown factor infertility while free thiols were significantly higher (P < 0.01) in unknown infertility factor. No significant difference was observed between IVF success/failure in any of the seminal parameters studied. There was a tendency for protamine deficiency to be higher and disulphide concentration to be lower in men withHRG633T. Such assessments may provide additional useful information about the prognosis for ART outcome, although more research is needed before clinical guidelines can be provided.

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Effect of long-term storage in Safe Cell+ extender on boar sperm DNA integrity and other key sperm parameters

Acta Veterinaria Scandinavica ◽

10.1186/s13028-017-0325-9 ◽

2017 ◽

Vol 59 (1) ◽

Cited By ~ 7

Author(s):

Wiesław Bielas ◽

Wojciech Niżański ◽

Agnieszka Partyka ◽

Anna Rząsa ◽

Ryszard Mordak

Keyword(s):

Sperm Parameters ◽

Dna Integrity ◽

Long Term Storage ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Boar Sperm ◽

Term Storage

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823 Prognostic value of sperm DNA integrity for ART (assisted reproductive technique) success: Empirical antioxidant therapy as a method of sperm DNA fragmentation and ART failure correction

European Urology Supplements ◽

10.1016/s1569-9056(16)60825-4 ◽

2016 ◽

Vol 15 (3) ◽

pp. e823

Author(s):

M.N. Korshunov ◽

E.S. Korshunova ◽

M.Y. Gabliya ◽

L.B. Kindarova ◽

J.A. Shtyrya

Keyword(s):

Dna Fragmentation ◽

Prognostic Value ◽

Antioxidant Therapy ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Assisted Reproductive Technique ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Reproductive Technique

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Sperm DNA integrity status is associated with DNA methylation signatures of imprinted genes and non-imprinted genes

Journal of Assisted Reproduction and Genetics ◽

10.1007/s10815-021-02157-6 ◽

2021 ◽

Author(s):

Bing Song ◽

Chao Wang ◽

Yujie Chen ◽

Guanjian Li ◽

Yang Gao ◽

...

Keyword(s):

Dna Methylation ◽

Dna Integrity ◽

Imprinted Genes ◽

Sperm Dna Integrity ◽

Sperm Dna

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Sperm DNA fragmentation: causes and identification

Zygote ◽

10.1017/s0967199419000595 ◽

2019 ◽

Vol 28 (1) ◽

pp. 1-8 ◽

Cited By ~ 1

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.

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Evaluation of zearalenone andα‐zearalenol toxicity on boar sperm DNA integrity

Journal of Applied Toxicology ◽

10.1002/jat.1322 ◽

2008 ◽

Vol 28 (5) ◽

pp. 681-688 ◽

Cited By ~ 24

Author(s):

Ioannis A. Tsakmakidis ◽

Aristoteles G. Lymberopoulos ◽

Tarek A. A. Khalifa ◽

Constanten M. Boscos ◽

Aikaterini Saratsi ◽

...

Keyword(s):

Dna Integrity ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Boar Sperm

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In Vitro Studying the Effect of Adding Autologous Platelet Rich Plasma (PRP) to the Human Semen on the Sperm DNA Integrity

Iraqi Journal of Embryos and Infertility Researches ◽

10.28969/ijeir.v10.i2.r7 ◽

2021 ◽

Vol 10 (2) ◽

pp. 90-100

Author(s):

Dhafer Hamdan ◽

Ali Rahim ◽

Ula Al-Kawaz

Keyword(s):

Dna Fragmentation ◽

Assisted Reproductive Technologies ◽

Platelet Rich Plasma ◽

Reproductive Technologies ◽

Human Reproduction ◽

Sperm Chromatin ◽

Dna Integrity ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Dispersion Test

For conception and the development of healthy embryos, sperm DNA integrity is crucial. According to a growing body of studies, there is a strong correlation between sperm DNA damage and male infertility. Among the new medicines being developed in the medical field, the application of Platelet Rich Plasma (PRP) in human reproduction has yet to be examined. A total of 100 semen samples were used in the current experimental investigation. From November 2020 to June 2021, the research was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Masturbation was used to get an ejaculated semen sample. After semen analysis, the samples were separated into two equal parts, one without autologous PRP and the other with 2% autologous PRP, with the DNA fragmentation assessed using the Sperm Chromatin Dispersion Test. There was highly significant reduction in DNA fragmentation index (p < 0.001). The mean sperm DNA integrity was reduced after adding PRP (33.85±16.73 vs 38.55±16.64), Mean (± SE). PRP has been shown to improve human sperm DNA integrity.

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Influence of bovine sperm DNA fragmentation and oxidative stress on early embryo in vitro development outcome

Reproduction ◽

10.1530/rep-13-0123 ◽

2013 ◽

Vol 146 (5) ◽

pp. 433-441 ◽

Cited By ~ 48

Author(s):

Renata Simões ◽

Weber Beringui Feitosa ◽

Adriano Felipe Perez Siqueira ◽

Marcilio Nichi ◽

Fabíola Freitas Paula-Lopes ◽

...

Keyword(s):

Oxidative Stress ◽

Dna Fragmentation ◽

Sperm Chromatin ◽

Dna Integrity ◽

Sperm Dna Fragmentation ◽

Cleavage Rate ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Significant Difference

Sperm chromatin fragmentation may be caused by a number of factors, the most significant of which is reactive oxygen species. However, little is known about the effect of sperm oxidative stress (OS) on DNA integrity, fertilization, and embryonic development in cattle. Therefore, the goal of this study was to evaluate the influence of sperm OS susceptibility on the DNA fragmentation rate and in vitro embryo production (IVP) in a population of bulls. Groups of cryopreserved sperm samples were divided into four groups, based on their susceptibility to OS (G1, low OS; G2, average OS; G3, high OS; and G4, highest OS). Our results demonstrated that the sperm DNA integrity was compromised in response to increased OS susceptibility. Furthermore, semen samples with lower susceptibility to OS were also less susceptible to DNA damage (G1, 4.06%; G2, 6.09%; G3, 6.19%; and G4, 6.20%). In addition, embryo IVP provided evidence that the embryo cleavage rate decreased as the OS increased (G1, 70.18%; G2, 62.24%; G3, 55.85%; and G4, 50.93%), but no significant difference in the blastocyst rate or the number of blastomeres was observed among the groups. The groups with greater sensitivity to OS were also associated with a greater percentage of apoptotic cells (G1, 2.6%; G2, 2.76%; G3, 5.59%; and G4, 4.49%). In conclusion, we demonstrated that an increased susceptibility to OS compromises sperm DNA integrity and consequently reduces embryo quality.

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