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Reproductive and epigenetic parameters in sperm connected to the disruptive development and early embryo loss.
Flintová, Jennifer ; Hortová, Kateřina (advisor) ; Petelák, Aleš (referee)
DNA methylation, histone modifications and regulation by non-coding RNAs are considered to play vital role in embryonal development and gametogenesis. Epigenetic mechanisms are not only inwardly programmed, but are massively affected by numerous exogenous aspects. Environmetal pollution and unhealthy parental lifestyle are two major factors related to impaired fertility status, abberant embryonal development of progeny. Some altered epigenetic marks can be transmitted to offspring generations, however in some cases these aberrations may be reversed by adequate lifestyle interventions. Since epigenetic regulations and genes responsible for spermatogenesis are conserved among vertebrates, studies based on animal models are highly relevant for understanding mechanisms causing impaired fertility and overall health in humans. Key words: Epigenetics, sperm, endocrine disruptors, hypoxia, lifestyle, transgenerational effect.
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The effect of diabetes mellitus on reproductive parameters and expression of selected testicular genes in diabetic mice
Valášková, Eliška ; Pěknicová, Jana (advisor) ; Kaňka, Jiří (referee)
According to the World Health Organization (WHO), 15% of couples in reproductive age suffer from infertility problems, and up to 60% of cases are caused by male factor. Causes of this condition could be genetic background, environmental factors and various diseases, including diabetes mellitus (DM). The aim of this study was to investigate the effects of DM on reproductive parameters and expression of selected testicular genes using mouse model (FVB inbred mouse strain). DM (type 1) was artificially induced by chemical substance streptozotocin, which causes destruction of pancreatic β cells. These mice were exposed to diabetic condition for 6 weeks and then subjected to analysis. Our results have shown that diabetic condition had an impact on body weight, weight of reproductive organs as well as kidneys and livers. We also observed decreased concentration and viability of diabetic sperm compared to control. Moreover, we noticed increased staining with apoptotic marker annexin V. Further, we evaluated changes of sperm nuclear proteins - protamines. In diabetic animals, we observed higher number of sperm with insufficient protamination. Nevertheless, protamine 1 to protamine 2 ratio (P1/P2), a marker of male fertility, was not altered in sperm of diabetic animals compared to control. Regarding the...
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Sperm morphology and molecular mechanism associated with sperm fertilizing ability in mice.
Šebková, Nataša ; Hortová, Kateřina (advisor) ; Jonáková, Věra (referee) ; Linhart, Otomar (referee)
The process of sperm capacitation and acrosome reaction (AR) are highly dynamic processes essential for the fertilization, including cytoskeleton proteins in the sperm head. The study of the distribution of actin, spectrin and α-tubulin in the head of representative mammalian sperm indicates that before and after the AR there were changes in the distribution of the cytoskeleton structures in the sperm head of all representatives, mainly in the apical part of the acrosome, but also in the equatorial and postacrosome part of the sperm head. The particularly well-studied actin cytoskeleton plays an important role during the capacitation and before and after the AR. Environmental estrogens can interfere with the function of endogenous hormones in very low concentrations. They can interact with specific receptors affecting several signalling pathways leading to sperm capacitation and AR. The effect of 17β-estradiol, estrone, estriol and synthetic 17α-ethynylestradiol was evaluated by the ratio of tyrosine phosphorylation and the state of the acrosome during in vitro capacitation in the sperm head. This study has provided the evidence that estrogens significantly stimulate capacitation progress in a concentration-dependent manner. Estrogens decrease number of sperm after the induced AR too. The raising...
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Výživa plemenných býků
KOUBA, Jindřich
The aim of this thesis was the study of available publications covering the topic of breeding bulls nutrition, the analysis of the feed dose and the evaluation of the semen quantity and quality after addition of extruded flaxseed (EL) to the feed dose. 200 semen samples from 8 breeding bulls of the Czech Fleckvieh were proposed as the subject of observation. The influence of EL to the sperm motility and concentration and to the amount of semen was observed and evaluated. The positive effect was found out in all of the observed characteristics. Experimental group showed increase in the amount of semen in comparison to the control group, and the statistical analysis displayed a significant difference in the effect of the time factor (effect of the addition of EL) on the amount of semen within the groups. The sperm concentration in 1 mm3 of semen of the experimental group increased compared to the control group, but the difference was not statistically significant. The experimental group showed an increase in sperm activity and the time factor was statistically significant. From the results of this thesis follows that extruded flaxseed can be recommended as the suitable feed supplement for breeding bulls.
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The influence of estrogens on mouse sperm capacitation and acrosome reaction in vitro
Tejnická, Magda ; Komrsková, Kateřina (advisor) ; Linhart, Otomar (referee)
There are an increasing amount of compounds in the environment that can have a negative effect on reproductive parameters in both male and female organism. There has been a worldwide decline of sperm quality during past decades and this fact lead to an increase of unnatural ways of conception through assisted reproduction techniques in the specialised centres. Natural estrogens are one of these compounds and they get into waste water after being excluded from the body by the urine. They get back into the human body from drinking water or from the food, and they can interfere with function of endogenous hormones in very low concentrations. For these reasons it is up to date to deal with the influence of these compounds on mammalian sperm. For many years, estrogens have been considered typically female sex hormones. It is now certain that they are also very important in the regulation of male reproduction. Endogenous estrogens in mammalian males are an important part of the endocrine system. Estrogens play an important role in the development of germ cells, spermatogenesis and processes leading to successful egg fertilization such as a capacitation or acrosomal reaction. Tyrosine phosphorylation is one of the essential steps for the properly ongoing process of capacitation in sperm followed by a...
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Analysis of chromosomal aberrations in sperm by fluorescence in situ hybridization
Bendová, Petra ; Diblík, Jan (advisor) ; Novotná, Drahuše (referee)
The presented bachelor work is focused on the determination of frequency chromosomally abnormal sperm in the semen of healthy men (donors) with normal karyotype (46, XY). The important process, which plays an irreplaceable role in the development of numerical aberrations of chromosomes or structural abnormalities in the segregation of the gametes, is meiosis. Therefore, I devote much attention on meiosis in the theoretical part. The theoretical part is focused on the process of pre mature sperm (spermatogenesis), and the consequences of fertilize the oocyte by aneuploid sperm. In my work I present an overview of numerical abnormalities in autosomes and gonosomes and their frequency and distribution of gametes in healthy men. I also focused on the distribution and a brief description of structural aberrations affecting chromosomes and not least I paid attention on method of multicolor interphase fluorescence in situ hybridization, which in combination with sperm chromatin dekondenzation become irreplaceable and valuable research tool for rapid analysis of chromosomal abnormalities in large sperm samples. The experimental part of bachelor work deals with monitoring the frequency of selected numerical abnormalities in sperm samples of five donors aged 23 to 30 years with the use of I-FISH (fluorescence in situ...
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Isolation and characterisation of oviductal proteins involved in formation of sperm oviductal reservoir in cow
Kračmerová, Jana ; Liberda, Jiří (advisor) ; Postlerová, Pavla (referee)
The oviduct plays an important role in the complex process of fertilization and in the early phase of embryo development. Isolation and characterization of some components of bovine oviduct participating in the formation of oviductal sperm reservoir was a subject of the present study. Two affinity sorbents containing immobilized components of bull seminal plasma (non-separated proteins of bull seminal plasma and their phosphorylcholine binding fraction) were prepared and used for the study of interaction of immobilized ligands with components of oviductal epithelium. Two types of preparation of oviductal membrane proteins of the isthmus region were isolated: complete membrane proteins and the apical fraction of membrane proteins. These preparations were separated using prepared affinity matrices. in both cases only an insignificant amount of proteins was found in the adsorbed fractions. Further attention has been paid to the identification of four protein zones detected after electrophoretic separation in the presence of SDS (rel. mol. wt. 15 500 - 18 500) in the fraction of apical membrane proteins. These proteins were identified after the electrophoretic separation and tryptic digestion using MS analysis as bovine histones H4, H2A typ 2-C, H2B typ 1-K a H3.3. Proteins identified as bovine...
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