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Meiotic sex chromosome inactivation within mouse spermatogenesis
Homolka, David ; Jansa, Petr (advisor) ; Kubelka, Michal (referee) ; Pěknicová, Jana (referee)
Meiotic sex chromosome inactivation (MSCI) is an essential epigenetic process, which transcriptionally silences X and Y chromosomes during spermatogenesis. It is accompanied by substantial chromatin remodeling resulting in a formation of so called sex or XY body, which is a characteristic of male pachytene spermatocytes. In spite of MSCI indispensability for male fertility, its biological role and molecular nature still remain rather unclear. However, the described link between chromosomal asynapsis and transcriptional silencing demonstrated that MSCI is tightly associated with the asynapsis of largely non-homologous sex chromosomes and is a specific form of more general mechanism called meiotic silencing of unsynapsed chromatin (MSUC). The essential role of MSCI was demonstrated using mouse models, such as carriers of X- autosome translocations, where anomalous synapsis of sex chromosomes leads to impairment of MSCI and male sterility. Intriguingly, the exclusive spermatogenic arrest is a hallmark of not only X-autosome translocations but even various autosomal rearrangements, including autosomal translocations, inversions, or other structural mutations. Because the rearranged autosomes often intimately associate with the sex body, it...
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Karyotype analysis of selected groups of tetrapulmonate arachnids
Reyes Lerma, Azucena Claudia ; Sember, Alexandr (advisor) ; Johnson Pokorná, Martina (referee) ; de Bello Cioffi, Marcelo (referee)
Arachnids are a highly speciose group of arthropods with about 130 000 extant species from 12 orders. Some arachnid species are of economic and medical interest. They also offer various opportunities to study the link between genome and karyotype variability and morphological and ecological diversification, evolution and adaptation. The knowledge about arachnid karyotypes and genomes is, however, still limited. The available data show wide range of diploid chromosome numbers (2n), monocentric or holocentric chromosomal structure, variability in sex chromosome systems (especially in spiders) and modifications of meiosis. The main objective of this doctoral thesis was to study by conventional and molecular cytogenetic methods the mechanisms of karyotype evolution in haplogyne spiders (with monocentric and holocentric chromosomes), and further in two families (Charinidae and Phrynidae) from the closely related order Amblypygi. The selected groups offer unique insights into fundamental karyotype traits of the arachnid clade Tetrapulmonata, and they also enable to analyse the role of polyploidy in arachnid karyotype evolution and the evolutionary mechanisms behind the emergence of holocentric chromosomes. Our data suggest that holocentric chromosomes are an autoapomorphy for the haplogyne superfamily...
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Male infertility and DNA germ cell breaks affected by the epigenetic factor PRDM9
Kusari, Fitore ; Trachtulec, Zdeněk (advisor) ; Děd, Lukáš (referee) ; Liška, František (referee)
DNA-binding histone-3-lysine-4,36-trimethyltransferase PRDM9 specifies meiotic recombination hotspots in mice, rats and humans. Interallelic variation at the Prdm9 locus plays a role in hybrid male sterility. Sterile mouse F1 hybrid male offspring from the PWD × C57BL/6J (B6) cross exhibit meiosis breakdown reminiscent of that observed in Prdm9-deficient B6 sterile mice. However, reciprocal (B6 × PWD)F1 hybrids and some rodent models lacking PRDM9, i.e., PWD and SHR rat males execute meiotic recombination, produce sperm, raising the possibility that PRDM9's role may extend beyond meiosis. Here I demonstrate that PRDM9 is important for post- meiotic male gamete development and release. Unlike their parents, (B6 × PWD)F1 generated spermatozoa of lower quantity and motility but higher percentage of deformations, thus resembling oligoasthenoteratozoospermic (OAT) (semi)sterile men. Histopathological and (ultra)structural analysis revealed compromised spermiogenesis characterized by acrosome detachment and aberrant nucleus elongation in (B6 × PWD)F1 hybrids. Consequently, F1 spermatozoa had malformed acrosomes and nuclear DNA breaks with elevated base oxidation. While deletion of one Prdm9 copy improved sperm phenotypes in (B6 × PWD)F1, copy number gains of the surrounding genes had the opposite effect....
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Gametogenesis and Fertilization in Humans
Crhová, Michaela ; Pavlasová, Lenka (advisor) ; Ehler, Edvard (referee)
This bachelor thesis summarises the topic of gametogenesis and fertilization in humans. A search in available literature was performed. The thesis concisely describes the cell cycle and cell division. The main focus is on meiosis, a reductional division that takes the lead part in sexual reproduction. The process of this division is described in detail, and also the mechanism that creates variability of protected cells is described. The product of meiosis is gametes, also known as germ cells. Production of these cells is a key moment in sexual reproduction. This thesis approaches this topic in the context of evolutionary biology and explains its importance in the survival of species on Earth. The main topic of the thesis is the production and maturing of human germ cells. This process is called gametogenesis. This thesis provides a cohesive description of the reproduction system of both sexes and describes the gradual maturing of the germ cells in gonads. The structure of both gametes, egg, and sperm is described with emphasis on the parts of these cells that take part in the process of fertilization. Female reproductive cycles are depicted, and their connection is explained. In contrast to this periodicity, the male continual maturing is also explained. This thesis compares the main differences in...
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Earlier evolutionary dead-ends, now the creators of a reproductive strategy: the origin and reproduction of the all-male water frog lineage Pelophylax esculentus
Kaštánková, Marie
Asexual modes of reproduction are usually based on the principle of copying (cloning) DNA from the female and passing it on to the offspring. For most asexually reproducing vertebrates the progeny develop from an unreduced and often unfertilised egg. This is driven by the mechanisms of parthenogenetic and gynogenetic reproduction. While in the former the clonal germ cell develops spontaneously and separately, in the latter a sexual partner is needed to activate the cleavage of the ovum, although without the fusion of the sperm and egg. Therefore in both cases there is no fertilization and the clonal progeny consist solely of daughters, hence the majority of previous studies have only focused on asexual female lineages. However, on rare occasions asexual clonal males can arise when the right fertilization occurs. Whilst these offspring are usually infertile, fertile diploid asexual males have been discovered in just three genera of hybrid origin in vertebrates. One of these unique cases is the European water frog complex of the genus Pelophylax, whose distribution includes the Czech Republic. In areas around the upper Odra River populations of hybrid males were recently discovered who form stable all-male lineages, similar to those formed by asexual females. The results of this study show that males produce...
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Regulation of mTOR pathway in the oocyte meiosis
Schmidtová, Barbara ; Šušor, Andrej (advisor) ; Krylov, Vladimír (referee)
Mammalian Target of Rapamycin (mTOR) is serin-threonin kinase, which has become a major topic in many studies in the last decade, leading to new insights into how cell works. This kinase is involved in proteosynthesis, metabolism, cell cycle regulation, proliferation and responses to nutrients and growth factors. There are certain diseases caused by mutations in mTOR gene, which lead to abnormal function of this kinase. These diseases include cancer or fertility disorders. mTOR research is also beneficial due to the search for drugs that could rescue its function and thus provide treatment for these diseases. The best-known drug is inhibitor Rapamycin and its derivates. The aim of this bachelor thesis is to summarize the knowledge about how mTOR can be regulated, the role of its substrates in cell function and to define the role of mTOR in oocyte development, translation and human health. Keywords: Oocyte, mTOR, translation, 4E-BP1, MPF, meiosis, AKT
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Genomic architecture and molecular mechanisms of hybrid sterility in mice.
Vališková, Barbora ; Forejt, Jiří (advisor) ; Janko, Karel (referee) ; Macholán, Miloš (referee)
Hybrid sterility is one of the reproductive isolation mechanisms restricting gene flow between the related species and leading to speciation. PR domain containing 9 (Prdm9), the only known vertebrate hybrid sterility gene, determines the sites of programmed DNA double-strand breaks (DSBs) and thus specifies hotspots of meiotic recombination but in hybrids between two mouse subspecies causes failure of meiotic chromosome synapsis and hybrid male sterility. In the present study on sterile hybrids, the five smallest autosomes were more prone to asynapsis. To manipulate with the synapsis rate, random stretches of consubspecific homology were inserted into several autosomal pairs. Twenty seven or more megabases of consubspecific sequence fully restore synapsis in a given autosome. Further, at least two symetric DN double-strand breaks per chromosome were necessary for successful synapsis. Moreover, F1 hybrids had sperm when synapsis was rescued in at least three of four segregating chromosomes. To verify the assumption of a lack of symmetric DSBs in meiotic chromosomes of sterile males the chemotherapeutic drug cisplatin was used to induce exogenous DNA DSBs. Cells treated with 5 mg/kg and 10 mg/kg of cisplatin showed increased number of DSBs monitored by immunostaining of RPA and DMC1 sites and...
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Genomics and cell biology of oxymonads
Treitli, Sebastian Cristian ; Hampl, Vladimír (advisor) ; Brune, Andreas (referee) ; Beneš, Vladimír (referee)
Oxymonads are a group of poorly studied protists living as intestinal endosymbionts in the gut of insects and vertebrates. In this thesis I focused on the study of phylogeny, genomics and cell biology of oxymonads. Using culture-based approaches, we uncovered the hidden diversity of small oxymonads and described one new genus and six new species. In Monocercomonoides exilis, the only oxymonad with a published genome, we investigated the genome organization using fluorescence in situ hybdridization (FISH) against the telomeric regions and single-copy genes. Our results show that the genome is most probably haploid being organized in 6-7 chromosomes. Annotation of the genome revealed that the DNA replication and repair mechanisms in M. exilis are canonical and they seem more complete than those of other metamonads whose genomes are available. Although M. exilis lacks in any traces of mitochondria, its genome annotation revealed that other cellular systems do not markedly differ from other eukaryotes. Our taxon-rich phylogenetic analyses suggested that the genus Monocercomonoides is closely related to the oxymonad Streblomastix strix, which is found exclusively in the gut of the termites. Streblomastix strix, as opposed to M. exilis, is highly adapted to harbour bacterial ectosymbionts. Since S. strix...
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Meiosis and fertility of juvenile mouse males
Valtrová, Pavlína ; Trachtulec, Zdeněk (advisor) ; Reifová, Radka (referee)
This work is a summary of literature on pecularities of spermatogenesis in juvenile mouse males (Mus musculus) and their utilization. Spermatogenesis is a process that leads through meiosis to sperm production. The cell undergoes in waves the following cell types: spermatogonia, spermatocytes, spermatids, and spermatozoa. Juvenile mice (whose testes size and sperm count have not reached their maximum) are often used to study individual cell types. The transition between cell types takes shorter time in juveniles. Spermatozoa from the 1st wave of spermatogenesis (WS) are derived from prenatal gonocytes, allowing earlier sperm production. They have a lower frequency of crossing over (CO rate) due to a different processing of CO intermediates; the consequence can be aneuploidy (one chromosome less/more). Spermatozoa from the 2nd WS still display lower CO rate. In 3rd WS testes descend and their temperature decreases to 33řC; CO rate is more like in adults. In 4th WS is typical testicular supportive cells mature and CO rate is similar to adult levels. Juvenile males also suffer from more frequent and severe sperm malformations. Low CO rate should not have an impact on fertility; errors are eliminated during meiotic checkpoints. However, the children of young fathers have a higher risk of aneuploidy,...
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Earlier evolutionary dead-ends, now the creators of a reproductive strategy: the origin and reproduction of the all-male water frog lineage Pelophylax esculentus
Kaštánková, Marie
Asexual modes of reproduction are usually based on the principle of copying (cloning) DNA from the female and passing it on to the offspring. For most asexually reproducing vertebrates the progeny develop from an unreduced and often unfertilised egg. This is driven by the mechanisms of parthenogenetic and gynogenetic reproduction. While in the former the clonal germ cell develops spontaneously and separately, in the latter a sexual partner is needed to activate the cleavage of the ovum, although without the fusion of the sperm and egg. Therefore in both cases there is no fertilization and the clonal progeny consist solely of daughters, hence the majority of previous studies have only focused on asexual female lineages. However, on rare occasions asexual clonal males can arise when the right fertilization occurs. Whilst these offspring are usually infertile, fertile diploid asexual males have been discovered in just three genera of hybrid origin in vertebrates. One of these unique cases is the European water frog complex of the genus Pelophylax, whose distribution includes the Czech Republic. In areas around the upper Odra River populations of hybrid males were recently discovered who form stable all-male lineages, similar to those formed by asexual females. The results of this study show that males produce...
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