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Effect of sperm ubiquitination on the early embryonic development in pig
Kroumanová, Kristýna ; Krylov, Vladimír (advisor) ; Nevoral, Jan (referee)
The ubiquitin-proteasomal system which is the major pathway for intracellular protein degradation is also involved in sperm quality control in the mammalian epididymis. Defective sperm become surface- ubiquitinated during epididymal passage. The level of sperm surface ubiquitination negatively correlates with their quality. Hypothetically it is possible that after fertilization, highly ubiquitinated sperm, naturally present in mammalian ejaculates, would be actively recognized by oocyte (probably via 26S proteasomal complex). Subsequent partial or total sperm degradation should negatively affect the development of the potentially defective embryo. In this study, we examined the effect of sperm ubiquitination on the early embryonic development in pig (Sus scrofa f. domestica) using the method of intracytoplasmic sperm injection (ICSI). In vitro embryonic development to the blastocyst stage after ICSI was comparable with other laboratories. In this study, no significant difference was observed in the formation of pronuclei between oocytes fertilized by lower and highly ubiquitinated sperm cells. On the other hand, significantly better embryonic development to the blastocyst stage was observed in oocytes fertilized by sperm with lower surface ubiquitination (17 %) compared with oocytes fertilized by highly...
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Dlx genes in odontogenesis and craniofacial mophogenesis in bichirs
Macháčová, Simona ; Černý, Robert (advisor) ; Zahradníček, Oldřich (referee)
Bichirs, Polypteriformes, are a basal group of vertebrates with many unique characters as ganoid scales, fleshy pectoral fin, spiraculum or paired lung originated from ventral outpocketing from floor of pharynx. Among the most notable is also the dentition, which contatin teeth among whole area of oropharyngeal cavity. This arrangement is not easily call oral and pharyngeal, what is recently used e.g. for medaka, because it is only one dention with teeth on majority of oral as well as pharyngeal elements. This diploma thesis utilizes expression patterns of Dlx genes to visualize morphogenesis of dentition and pharyngeal arches in the Senegal Bichir (Polypterus senegalus). The very first developing teeth of oral dentition, as well as its further patterning were detected and accurately located using gene expression analyses. It was also discovered an interesting histological context connected with unique mouth opening mechanism during early development of teeth in oral cavity. Another discussion point is establishment of dorsoventral polarization in pharyngeal arches development of bichir based on results from Dlx gene expression analysis. Powered by TCPDF (www.tcpdf.org)
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Physiological role of SIGIRR in early embryonic development.
Hanusová, Zdeňka ; Filipp, Dominik (advisor) ; Brdička, Tomáš (referee)
IL-1 receptor/Toll-like receptor (IL-1R/TLR) supefamily represents a group of proteins that share highly conserved TIR domain in their cytoplasmic region. Signal transduction mediated by TIR-containing proteins involves the activation of NF-κB transcription factor and thus the members of this superfamily play a key role in many physiological responses related to innate immune defense and inflammation. SIGIRR (single immunoglobulin IL1R-related molecule) is a recently discovered member of the IL-1R family, however it differs from the other group members by its unique structural features. SIGIRRhas been so far considered to be an 'orphan' receptor as no SIGIRR ligand has been identified yet. Moreover, SIGIRR itself is not capable to induce the NF-κB activation. Instead, SIGIRR is supposed to act as a negative regulator for IL- 1Rs/TLRs mediated inflammation. Its inhibitory function has been implemented in several signalling pathways in various cell types and tissues including the kidney, the digestive tract and the lung. Recent reports also suggest that SIGIRR could play a role in early embryonic development. The main aim of this thesis is to characterize the mechanism how SIGIRR negative regulatory function in IL-1R/TLR signalling pathway is delivered. Here we describe the establishment of...
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Formation of spatio-temporal molecular gradients in early embryonic development of Xenopus laevis.
Šídová, Monika ; Tlapáková, Tereza (advisor) ; Pěknicová, Jana (referee) ; Petr, Jaroslav (referee)
Clarifying the underlying spatio-temporal mechanisms that determine body pattern is important for detailed understanding of embryonic development. A crucial question of vertebrate embryogenesis remains: when and how are single blastomeres determined for differentiation that subsequently leads to body axes specification and the formation of different tissues and organs? The answer to this question will be beneficial for primary research as well as in the field of applied medicine. The main aim of the presented thesis was to study spatio-temporal molecular gradients of cell fate determinants during early embryonic development. The African clawed frog Xenopus laevis was used as a model organism because of their large size of oocytes and external embryonic development. Due to late activation of embryonic transcription, a crucial mechanism of early blastomeres determination is dependent on asymmetric localization of maternal factors within oocyte and their uneven distribution into single blastomeres during early cell division. Two main localization patterns were identified along the animal-vegetal axis of the mature Xenopus oocyte using qPCR tomography. The localization gradient with preference in either animal or vegetal hemisphere was found for maternal mRNA as well as miRNAs. Moreover, two vegetal...
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Some factors influencing success of IVF-embryo transfer
Pertoldová, Stanislava ; Rajmon, Radko (advisor) ; Marie, Marie (referee)
Assisted reproduction is a field of medicine that deals with infertility tretment.
Ongoing improvements of laboratory technology as well as ongoing evolving scientific research within IVF research,including experimental embryology in routine practice is contributing in a steady improvement of success rates of assisted reproduction.
This thesis is primarily based on data and information provided by the laboratory of assisted reproduction of GEST IVF Ltd.clinic in Prague,which was collected during the year 2015. This data was analyzed and investigated to verify the effect of selected factors on the success of the treatment. In This thesis we have not used any personal data of patients in accordance with the legislation act.296/2008Sb.
The results show clear evidence between the treatment success rate and patients age within all evaluated parameters. The fundametal hypothesis of interdependence of embryo quality on the further development of the embryo after the transfer was comfirmed.
Conversely, the timeframe for embryo cultivation is not an important parameter of success. Commonly used cuture media also did not show a significant impact on treatment outcome.
Statistical calculations also confirmed surprisingly significant differences in embryo transfer after 72 hours development and 120 hours development for women ever 35 years. Transfers of younger embryos appears to be more convenient for women over 35 years.
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Regulation of diapause in the western corn rootworm (\kur{Diabrotica virgifera virgifera} LeConte)
HOUFKOVÁ, Kateřina
The thesis aims to optimize the methods of laboratory culture and to fill in the gaps in knowledge of D. virgifera virgifera ecophysiology. The experiments on embryology and development proved that the diapause of D. virgifera virgifera is of obligatory type and can be terminated by exposure to long-day conditions (20:4; photo : scotophase) and constant temperature of 25 °C. Approximately 10% of eggs completed the development to adults within 4 months. Besides numerous other environmental conditions that are discussed, temperature seems to be a key factor influencing longevity in this pest. Higher temperature of 25 °C prolonged survival by more than 20 days in 2015, compared to 22 °C in 2014.
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The application of biotests on Japanese medaka (Oryzias latipes) eggs for the screening assessment of cyanobacterial water blooms toxicity
SIKORA, Jiří
This thesis has two parts. In the first part there is described an optimal methodological process for screening tests used in subsequently. Fertilized fish eggs of Oryzias latipes were incubated in 6 tests with different numbers (from 1 to 6) with standard conditions in ISO water. In the tests, hatching performance and duration of embryonic development were investigated and the results were applied on screening tests. The other part of the thesis is aimed on the proof of potential toxic effects of water with cyanobacterial water bloom. The fertilized eggs of Oryzias latipes were embedded into the test in stage 6 to 8. Three samples of cyanobacterial biomass from free waterbodies with known species composition and microcystin {--} LR, YR and RR contents were tested. The hatching performance, duration of embryonic development, lethal and sublethal effects were monitored during the tests. The tests were performed according to the OECD 212. There were detected significant differences in hatching performance, duration of embryonic development and in some cases also in induction of deformities between the control group and the tested groups.
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