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Sperm DNA integrity is an essential factor for successful fertilization and proper pregnancy progression. The terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay is commonly used for the estimation of the DNA fragmentation index. Analysis of TUNEL-stained sperm is often performed by flow cytometry, an approach that allows high-throughput analysis but in which any morphological information is lost. In this study, results of an automated image cytometry estimation of TUNEL-stained sperms were presented. The results of visual counting and automatic analysis were closely correlated, indicating that image cytometry is suitable for such analysis and may be applied in a clinical setting. © 2017 International Society for Advancement of Cytometry [ABSTRACT FROM AUTHOR]

The effect of mutation Kit W-Y found in C57BL/6 mice on fertility, spermatogenesis, and early embryogenesis of mice have been studied. If heterozygotes Kit W /+ are crossed with wild-type mice, fertility decreases by 20%. Homozygotes Kit W-Y /Kit W-Y and compounds Kit W-Y /Kit Ssm are nonviable. The study of spermatogenesis in Kit W /+ mice has demonstrated a negative effect of this mutation on spermatocytes. Histological examination of the testes of mutant males has shown local empty spaces in seminal ducts. Electron microscopic examination of synaptonemal complexes have demonstrated desynapsis disturbance in some nuclei at the diplotene stage of meiotic prophase I. However, these disturbances do not cause a decrease in the number of fertilized oocytes/ova. The decrease in fertility is accounted for disturbances of early embryogenesis. In vivo and in vitro analyses of early embryogenesis have demonstrated that cleavage divisions are asynchronous in Kit W-Y /+ heterozygous embryos. Some of these embryos die before implantation, and others cleave more rapidly than wild-type embryos, which give them selective advantage during the postimplantation period of embryogenesis. The pattern of Kit W-Y expression during spermatogenesis and embryogenesis mimics potential human pathology, which makes these mutants an interesting and valuable object for genetics and developmental biology. [ABSTRACT FROM AUTHOR]

Introduction: Sperm motility is one of the major determinants of male fertility. Semen samples from more then 8029 men of different fertility status were obtained in Research Centre for Medical Genetics, RAMS. Cohort of asthenozoospermic men were examined for ultrastructural changes. Materials and Methods: 8029 semen samples were analyzed according to World Health Organization criteria. Electron microscopic studies in cohort of 98 infertile men with asthenozoospermia (a<25% or a+ b<50% (WHO 2010) were carried out. Results: Semen analysis showed that reduced sperm motility is the most common alteration in male infertility (88%). In 70% of cases low sperm motility is associated with abnormal sperm morphology. Electron microscopic studies revealed acrosome abnormalities (knobbed, ruffled and incomplete), head defects (abnormal chromatin condensation, decapitated), tail structural defects (peri-axonemal and axonemal abnormalities, lacking dynein arms), etc. Conclusion: Our study results suggest that reduced sperm motility concerns more complicated interactions involved in control of spermatogenesis, either directly or indirectly. The approach will next be identified by further investigations. [ABSTRACT FROM AUTHOR]

Objective: Recent studies have demonstrate the presence of correlation between herpes simplex virus (HSV) presence in ejaculate and male infertility. The role of cytomegalovirus (CMV) in fertility disorders is also discussed. At the same time pathogenesis of viral impact on male reproductive system is unclear. Resistance of spermatozoa to herpesviruses assumes that virus affects germ cells during its maturation in testis. The aim of the study was to investigate HSV and CMV effects on immature germ cells (IGC) going through different stages of development -- spermatogonia, spermatocytes, spermatides. Materials and Methods: Analysis of IGC population was performed in HSV- and CMV-infected sperm samples and on the model of testis herpesvirus infection in vitro. 134 sperm samples from infertile males, including 89 HSV-infected, 20 CMV-infected and 20 virus-free samples, were investigated by quantitative karyological method. The in vitro model of HSV and CMV infection was obtained from the testicular organotypic culture. Viability of spermatogonia, spermatocytes, spermatides and spermatozoa in infected and uninfected culture was assessed during 16 days of culturing. Results: In HSV-infected samples we found an increase in degenerative germ cells comparing with uninfected samples (13.2% vs 7.2%, p < 0.05), and additionally, in the case of high viral load, we observed the decrease in spermatides (80.9% vs 86.5%, p < 0.05) and spermatocytes I in diplotene (0% vs 0.43%, p < 0.05). The same changes were detected while comparing CMV-infected sperm samples: the amount of degenerative germ cells was higher (16.2% vs 7.2%, p < 0.05), and the concentration of spermatides was lower (78.3% vs 86.5%, p < 0.05) in the samples with high viral load compared with virus-free samples. Investigation of IGC in organotypic culture also revealed the decrease in spermatocytes and spermatides in infected testis from Day 4 of culturing. On Day 16, the viability of spermatocytes in uninfected culture was 51.5%, comparing with 16.9% (p < 0.001) in HSV- and 18.7% (p < 0.001) in CMV-infected culture. The similar values for spermatides comprised 68% vs 39.9% (p < 0.001) and 35.1% (p < 0.001). More over the decrease in spermatogonia was detected in presence of HSV (35.5%, p < 0.001) and CMV (36.8%, p < 0.001), comparing with 82.2% of vital cells in virus-free testis culture. The population of spermatozoa was not impacted during 16 days of infection. Conclusions: HSV and CMV can affect IGC at different stages of maturation. Meiotic cells (spermatocytes) and the cells that are formed as a result of meiosis (spermatides) are the most sensitive to herpesviruses. The decrease in IGC population is a possible factor for fertility disorders in males infected by herpesviruses. [ABSTRACT FROM AUTHOR]