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Molecular mechanisms of coronary vasculature development
Neffeová, Kristýna ; Kolesová, Hana (advisor) ; Neckář, Jan (referee)
The cardiovascular system is the first functional system that develops in vertebrates during embryonic development. Its irreplaceable function is the transport of nutrients and the removal of waste products. During the development the heart not only grows, but also acts as a pump that drives the blood circulation of the embryo. With advancing development, it is necessary to ensure an adequate supply of oxygen to the heart, for that reason coronary arteries are formed. Each cardiomyocyte is surrounded by at least one capillary, therefore the interaction between cardiomyocytes and endothelial cells plays an indispensable role in the proper functioning of the heart. Understanding, how cardiomyocytes and endothelial cells communicate, is essential for medical research in cardiac tissue regeneration. A number of factors involved in coronary development are described in the literature. However, these factors are described as separate signaling pathways, not as a system of mutually interacting mechanisms. The main goal of my bachelor thesis is to connect individual signaling cascades important in cardiomyocyte-endothelial cell communication and describe their interactions. The main factors overviewed are VEGF, Notch, PDGF, Angiopoietin and others. Factors function and signalization is reviewed in details....
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Formation of spatio-temporal molecular gradients in early embryonic development of Xenopus laevis.
Flachsová, Monika
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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Výživa drůbežího embrya
Zahnáš, Radek
Poultry meat is considered dietetic due to its low fat content and excellent digestibility. Thanks to very short fattening period, which is about 35 days, the consumption of poultry meat is increasing. The in ovo method can be used to nutrition of chicken before hatching, which saves the yolk sac nutrients between hatching and transport to the farm where feed is available to the chicken for the first time. Saved nutrients of the yolk sac are available to the chicken to develop the immune system instead of consumption for conservation purposes. The in ovo method can also be used for the early vaccination of chickens still in the egg, when the chickens did not come into contact with the pathogens. Another indisputable advantage of this method is the possibility of determining the sex of unhatched chickens.
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Gene expression in chicken embryo: micromanipulation and visualization methods
Bendová, Michaela ; Dvořák, Michal (advisor) ; Hirsch, Ivan (referee) ; Krylov, Vladimír (referee)
The aim of this work was to obtain better insight into the principles of cell structures and organs in the chicken embryo development. To reach this goal special methods of micromanipulations and visualization in vitro, ex vivo, ex ovo and in ovo were implemented and adjusted. These methods were used to study gene expression in neural crest development and eye development. In the course of long term research in our laboratory we observed that oncoprotein v-Myb influences the development of the neural crest and has the capacity to change natural cell fate. We performed a series of experiments to investigate v-Myb protein influence on neural crest cells differentiation, especially melanocyte lineage development, and its influence on gene expression in the neural crest. Therefore we focused on Gremlin 2 (PRDC), the gene upregulated by v-Myb in the neural crest. The established procedure of electroporation in ovo was adjusted to transfect cells of the developing eye and used to study gene expression during lens induction. The results obtained from chicken embryo experiments endorsed the study performed on mouse embryos. Futhermore, the electroporation technique was slightly modified for manipulations of the neural retina in the developing eye in ovo. Thereafter, the retinas were processed ex vivo and...
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Transcriptional regulation in the development of neurosensory cells in the inner ear
Vochyánová, Simona ; Pavlínková, Gabriela (advisor) ; Schierová, Michaela (referee)
To understand the pathophysiology of hearing loss, it is necessary to identify genes responsible for specification and differentiation of sensory cells and neurons from a common neurosensory progenitor. These factors include LIM-homeodomain transcription factor ISLET1, high-mobility group protein SOX2, and basic helix-loop-helix transcription factors ATOH1, NEUROG1 and NEUROD1. This study aims to map important factors in inner ear development and their interactions with specific focus on transcription factor NEUROD1 and its role in mouse neurosensory inner ear development and function. Key words: inner ear, transcriptional regulation, mouse model, targeted deletion, embryonal development, Neurod1
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Asymetric segregation of cell determinants in embryos with regulatory development
Chramostová, Kamila ; Krylov, Vladimír (advisor) ; Černý, Robert (referee)
Asymmetric segregation of cell determinants specific the cell fates in early embryonic development. In C. Elegans somatic and germ cells are separeted in early development. On the basis of the first five asymmetric divisions AB, MS, C, E, D, P4 are found. In this specifications, par genes and genes responsible for the specification of blastomers are involved. P granules are segragated to P4 cells. Xenopus as a representative of amphibians, belonging to a group with regulative embryogenesis , exhibit features of mosaic development like C. Elegans. Maternal RNAs in germ plasm determine germ cells already in oocyte. These RNAs are transported through two distinct pathways. METRO pathways is used for a transportation of mitochondrial cloud RNA. It is specific for Xcat-2, Xpat, XIsirt, Xdazl. Vg1 and VegT RNAs are involved in the second pathway called Vg1 . This RNA contribute to the specification of mesoderm and endoderm. Although mammals reveal regulative development they also exhibit some features of the mosaic embryos. As an example, in the mouse embryo, overexpressing cadherin leads to compaction at the 8 -cell stage. Then the cells of embryo are polarized. Polarized internal cells give rise to ICM. Conversely, the outer nonpolarized cells give rise to TE. Specific factor for TE is Cdx2, for ICM is...
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Formation of spatio-temporal molecular gradients in early embryonic development of Xenopus laevis.
Flachsová, Monika
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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The role of vent genes family in early development and brain development
Fabian, Peter ; Kozmík, Zbyněk (advisor) ; Němec, Pavel (referee) ; Balaštík, Martin (referee)
6 III ABSTRACT (ENGLISH) In chordates, the central nervous system (CNS) is derived from the dorsal part of gastrula. Induced dorsal part of the embryo - the neural plate - gives rise to the neural tube or primordial brain. The developing dorsal part of the embryo is shaped by BMP/Smad signaling from the ventral part. Using the basal chordate amphioxus, we show here the conserved evolutionary role BMP/Smad signaling in axial cell fate determination. Pharmalogical inhibition of BMP/Smad signaling induces dorsalization of Branchiostoma floridae (amphioxus) and Oryzias latipes (medaka) embryos and expansion of neural plate markers. We provide evidence for the presence of the positive regulatory loop within the BMP/Smad signaling network of amphioxus. Thus, our data suggest that early emergence of a positive feedback loop within the BMP/Smad signaling network may represent a crucial molecular event in the evolutionary history of the chordate cell fate determination. The dorso-ventral body axis formation is mediated by genes of the vent family, which are the direct targets of BMP/Smad signaling. The function of vent gene family in early development is relatively well known, however, its role in developing CNS is not yet clear. Therefore, we decided to manipulate vox transcription factor, a vent family member....
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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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