|
|
Using Sperm Transcriptome in Search for Novel Biomarkers of Male Infertility
Semyakina, Anastasiya ; Liška, František (advisor) ; Stopka, Pavel (referee)
Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically useful tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of...
|
|
|
Bacterial populations in mucosal tissues of the house mouse
Ptáčníková, Aneta ; Stopka, Pavel (advisor) ; Hampl, Vladimír (referee)
Microbiota becomes one of the most important subjects in biological research and numerous studies revealed that microbiota plays a broad spectrum of essential roles in different organisms. This master thesis focuses on the bacterial part of microbiota contained in mucosal tissues of wild house mice (Mus musculus musculus). Male and female samples were collected by nasal and oral cavity lavages, vaginal mucosa lavages and from urine and stool. We aimed to detect quantitative, qualitative and sex-specific differences in bacterial populations between mucosal tissues with particular focus on bacterial cycling in vaginal mucosa during the estrous cycles. Bacterial abundances were estimated by qPCR whilst bacterial diversity was detected by targeted metagenomic sequencing of the hypervariable region of the 16S rRNA gene. Significant differences were detected in bacterial abundances and alpha diversity between particular mucosal tissues. Stool samples contained the highest number of bacteria, while samples from the nasal mucosa and urine contained low amount of bacteria. The highest alpha diversity was discovered in stool samples, the least alpha diversity was found in the urine. Mucosal tissues also varied based on the bacterial composition on the level of particular genera. Detailed analysis of estrous cycles...
|
|
|
Conserved Mechanisms of Gene Expression Regulation by Nuclear Receptors.
Novotný, Jan Philipp ; Kostrouch, Zdeněk (advisor) ; Stopka, Pavel (referee) ; Lazar, Jozef (referee)
7 Abstract With the first appearance of life on Earth, organisms had to adapt to an ever-changing surrounding environment in order to survive. Since the emergence of metazoan multi- cellularity, subsets of cells could adapt to perform specific biological tasks beneficial to the whole organism, necessitating not only spatiotemporal regulation of gene expression during development, but also integration of tissue specific needs with overall organis- mal status. Within the set of evolutionary conserved regulatory systems, the family of nuclear receptor (NR) transcription factors stands out due to its high degree of evolu- tionary conservation, plasticity and uniqueness to the metazoan kingdom, regulating gene expression in response to, or in the absence of a ligand by genomic and non- genomic actions. With an increasing number of different compounds being recognized as ligands to NRs, it is now thought that ancient NRs were probably characterized by low ligand binding specificity, eventually serving as environmental sensors, integrating nutrient availability and gene expression at the base of metazoan evolution. Characteri- zation of the NR network in one of the simplest metazoan organisms, Trichoplax ad- haerens, revealed not only a functional network and sub-specialization of NR dependent gene regulation, but...
|
|
|
Using Sperm Transcriptome in Search for Novel Biomarkers of Male Infertility
Semyakina, Anastasiya ; Liška, František (advisor) ; Stopka, Pavel (referee)
Currently infertility affects 10 to 15 percent of couples. In nearly 50% of all cases male factor contributes to infertility of the couple. Majority of causes of male infertility remains unexplained. For this reason, finding simple and clinically usefµl tools for improving male infertility diagnostics can be important at present. Based on the analysis of sperm transcriptome, in this diploma thesis we aimed to find genes that show differential expression between normal and pathological sperm, this could provide information about molecular basis of male infertility, moreover, expression profile of such genes in sperm could be employed for noninvasive diagnosis of male infertility. This study was conducted by using 67 sperm samples, including 16 control sperm samples from fertile men. The samples of infertile patients were divided into 3 groups according to morphology and motility using results of standard sperm evaluation according to WHO. First group included 20 patients diagnosed with astenoteratozoospermia (low sperm motility and abnormal morphology), second group was 15 patients diagnosed with asthenozoospermia (normal morphology and low sperm motility), the third group comprised 16 samples from infertile patients with normal spermiogram. 16 control samples were from fertile men (conception of...
|
|
| |
|
|
Analysis of dosage effect of speciation gene Prdm9 on fertility of mouse hybrids
Flachs, Petr ; Trachtulec, Zdeněk (advisor) ; Stopka, Pavel (referee) ; Král, Jiří (referee)
(eng) The phenomenon of hybrid sterility represents one of the evolutionary mechanisms that enables speciation. Only a few speciation genes have been uncovered. The only one found in mammals is Prdm9 (PR-domain 9). Data in the literature on the involvement of Prdm9 in decreased fertility of various semifertile hybrid males of house mouse subspecies were scarce before the results of this thesis were completed, despite that such males are much more frequent in nature than the fully sterile ones. Utilizing a panel of genetic tools and a battery of phenotyping tests, this thesis shows a central role of Prdm9 in fecundity of hybrids, including many fertility disorders and age dependency. Both increasing and reducing the Prdm9 gene dosage significantly elevated fertility parameters. Surprisingly, even the allele that in one copy causes full hybrid sterility increased F1 hybrid fertility when present in multiple copies. The PRDM9 protein also plays a role in identifying the sites of meiotic recombination. This study also points out the principles of allelic competition in determination of the sites of preferred recombination (hotspots), which suggests a possible link between both previously described Prdm9 roles. This thesis summarizes a set of three logically interconnected publications with the ambition...
|
|
| |
|
| |
|
| |
|
| |
guest ::
RSS feed.