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Sperm DNA damage: detection methods used in clinical practice
Tušlová, Magdaléna ; Postlerová, Pavla (advisor) ; Koubek, Pavel (referee)
The current routine semen analysis, known as a spermiogram, is not able to provide a complete picture of male fertility as it does not routinely include an assessment of DNA integrity. This parameter is closely related to difficulties during conception and increased risk of miscarriage. This paper discusses the causes of sperm DNA damage, the detection of this damage by various methods and concludes with a discussion of the possibilities of incorporating these tests into clinical practice to improve the assessment and treatment of male fertility. Mechanisms of sperm DNA fragmentation include impaired spermatogenesis, oxidative stress and impaired apoptosis, which can be influenced by genetic, anatomical and external factors. The main part of this thesis was devoted to a review of methods for the detection of sperm DNA fragmentation, including sperm chromatin structure analysis (SCSA), sperm chromatin dispersion test (SCD), TUNEL method, single-strand DNA break recognition test (ISNT), comet assay and chromatin staining. Although some of these methods are frequently used, challenges remain in their standardization and full integration into clinical practice. However, the possibility of their routine use could bring significant advances in the diagnosis and treatment of male infertility. Among the...
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The role of LINC complex in male germinal cells
Kmoníčková, Věra ; Komrsková, Kateřina (advisor) ; Knop, Filip (referee)
LINC (linker of the nucleoskeleton and cytoskeleton) complex is substantial for the overall function of any nuclear cell. It is a bridge between the cytoplasmic and nuclear structures which mediates transmission of mechanical forces generated on the cytoplasmic side. The main objective of this thesis is the function of distinct variants of the LINC complexes in mammalian male germinal cells throughout their development. The nuclear facing side of the LINC complex is a key player in homolog pairing and rapid chromosome movements during meiosis. The cytoplasmic facing side is necessary for the sperm head formation, proper acrosome and nucleus attachment. Evenly, another complex is essential for the head-to-tail coupling as without this one the sperm cells are found decapitated. If the subunits of the LINC complex are defective, males are afflicted with different kinds of infertilities, which nowadays is one of the important issues of our society. Many things about the role and structure of the LINC complex has been discovered due to modern technologies. These up to date findings are summarized and discussed on the following pages.
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Post-Translational Modifications of Nuclear and Non-Nuclear Proteins in Spermatozoa
Řimnáčová, Hedvika ; Nevoral, Jan (advisor) ; Krapf, Dario (referee) ; Álvarez Rodríguez, Manuel (referee)
Posttranslational modifications of nuclear and nonnuclear proteins in spermatozoa Summary The number of couples who need the help of assisted reproductive technology (ART) has increased over the years. Approximately half of the cases are caused by male infertility, which is often diagnosed as idiopathic infertility. Therefore, the search for male fertility markers will improve male infertility diagnosis, thereby facilitating advanced sperm treatment and selection via ART. Posttranslational modifications (PTMs) of sperm nuclear and nonnuclear proteins are suitable candidates for such markers. The PTMs of protamines and histones reflect sperm chromatin maturity and its readiness for fertilization, and accordingly, they can predict the outcome of ART. However, the PTMs of nonnuclear proteins, including cytoplasmic, cytoskeletal, and membrane proteins, reflect the ability of sperm to undergo hyperactivation, capacitation, or acrosome reactions, which are processes essential for fertilization. We hypothesize that the PTMs of nuclear and nonnuclear proteins can reflect sperm quality and, thus, serve as a valuable marker in ART. Additionally, we suggest that the in vitro addition of hydrogen sulfide into the sperm-manipulating media improves sperm motility and viability via persulfidation. We used Western blot...
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Targeted next generation sequencing of candidate genes responsible for impaired spermatogenesis and male infertility
Daňková, Michaela ; Liška, František (advisor) ; Holá, Dana (referee)
Infertility is a widespread health problem, caused by the male factor in about half of all cases, and in about a half of the infertile men the cause is unknown. In a significant number of these men, genetic etiology is assumed. Current routine methods of laboratory diagnostics, which include karyotype examination, exclusion of mutations in the CFTR gene, and Y chromosome microdeletions, do not usually reveal the cause of infertility. That is why researchers' efforts aim at detecting mutations in other genes that are causing male infertility. In recent years, animal models have been used to identify many genes necessary for fertility. Based on these findings, 12 candidate genes have been selected (CAPZA3, CDC14B, CDC42, CNTROB, CSNK2A2, GOPC, HOOK1, HRB, OAZ3, ODF1, RIMBP3, SPATA16) that are essential for spermatogenesis. Mouse or rat mutants in these genes are primarily associated with oligoasthenoteratozoospermia, since they are involved in sperm morphogenesis. However, the phenotype spectrum may comprise also azoospermia. The purpose of the thesis was to determine the sequence of the afore mentioned genes in infertile men with impaired spermatogenesis and to reveal presence or absence of pathogenic mutations in these genes, using cDNA and genomic DNA from peripheral blood. The candidate genes were...
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The role of LINC complex in male germinal cells
Kmoníčková, Věra ; Komrsková, Kateřina (advisor) ; Knop, Filip (referee)
LINC (linker of the nucleoskeleton and cytoskeleton) complex is substantial for the overall function of any nuclear cell. It is a bridge between the cytoplasmic and nuclear structures which mediates transmission of mechanical forces generated on the cytoplasmic side. The main objective of this thesis is the function of distinct variants of the LINC complexes in mammalian male germinal cells throughout their development. The nuclear facing side of the LINC complex is a key player in homolog pairing and rapid chromosome movements during meiosis. The cytoplasmic facing side is necessary for the sperm head formation, proper acrosome and nucleus attachment. Evenly, another complex is essential for the head-to-tail coupling as without this one the sperm cells are found decapitated. If the subunits of the LINC complex are defective, males are afflicted with different kinds of infertilities, which nowadays is one of the important issues of our society. Many things about the role and structure of the LINC complex has been discovered due to modern technologies. These up to date findings are summarized and discussed on the following pages.
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Anthropological aspects of male infertility
Sakmárová, Františka ; Novák, Jan (advisor) ; Brynychová, Iva (referee)
Nowadays, more and more couples have had trouble with conceiving a child. Worldwide, around 15 % couples are affected by infertility, while men make up for 20-70 % of this problem. In last decades, the prevalence of the male infertility has been rising globally, with the sperm quality parametres becoming worse. From the view of male infertility etiology, it is rather difficult to pinpoint reasons of this trend. Besides sick individuals whose infertility is caused by genetic or congenital malfunctions or infections, the male infertility affects seemingly healthy individuals as well. The infertility of these objectively healthy (albeit infertile) individuals is from the etiological point of view said to be caused by environmental factors, worst of them being endocrine disruptors, and lifestyle factors, such as obesity, alcohol, smoking and stress. This thesis presents a complex view of male infertility problem, describing both its current state in epidemiology and its changes in time and space. Further on, the thesis is dedicated to the etiology of male infertility, including the lifestyle factors, where anthropology point of view is accentuated. Keywords: male infertility, reproduction, sperm, sperm parameters, causes of male infertility
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Male infertility in context of testicular cancer
Cimlerová, Markéta ; Komrsková, Kateřina (advisor) ; Truksa, Jaroslav (referee)
This bachelor thesis focuses on male infertility in a connection to testicular cancer. Testicular cancer is the most common malignancy among young men in a reproductive age and the worldwide incidence of testicular cancer is on the rise. A lot of attention is also given to an increasing rate of infertility in a context of testicular cancer. For these reasons, the aim of this thesis is to clarify several non-physiological changes, such as hormone levels, spermatogenesis and sperm parameters, which take place in a male body and influence the chance to become a biological father. These pathological changes can be due to the disease itself but also due to the treatment. A cryopreservation of semen is also going to be discussed as the way to preserve male fertility and as an option for couples with the infertility problems due to male factor which rely on help of the assisted reprodiction. Kye words: testicular cancer, cancer treatment, male infertility, sperm parameters, mitochondria, assisted reproduction
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Targeted next generation sequencing of candidate genes responsible for impaired spermatogenesis and male infertility
Daňková, Michaela ; Liška, František (advisor) ; Holá, Dana (referee)
Infertility is a widespread health problem, caused by the male factor in about half of all cases, and in about a half of the infertile men the cause is unknown. In a significant number of these men, genetic etiology is assumed. Current routine methods of laboratory diagnostics, which include karyotype examination, exclusion of mutations in the CFTR gene, and Y chromosome microdeletions, do not usually reveal the cause of infertility. That is why researchers' efforts aim at detecting mutations in other genes that are causing male infertility. In recent years, animal models have been used to identify many genes necessary for fertility. Based on these findings, 12 candidate genes have been selected (CAPZA3, CDC14B, CDC42, CNTROB, CSNK2A2, GOPC, HOOK1, HRB, OAZ3, ODF1, RIMBP3, SPATA16) that are essential for spermatogenesis. Mouse or rat mutants in these genes are primarily associated with oligoasthenoteratozoospermia, since they are involved in sperm morphogenesis. However, the phenotype spectrum may comprise also azoospermia. The purpose of the thesis was to determine the sequence of the afore mentioned genes in infertile men with impaired spermatogenesis and to reveal presence or absence of pathogenic mutations in these genes, using cDNA and genomic DNA from peripheral blood. The candidate genes were...
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