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Phenotypic study Huntington's disease TgHD minipigs: Appearance and progress of reproductive and biochemical changes
Bohuslavová, Božena ; Motlík, Jan (advisor) ; Roth, Jan (referee) ; Petr, Jaroslav (referee) ; Krylov, Vladimír (referee)
Huntington's disease (HD) is one of the incurable and fatal diseases. HD belongs to the monogenic neurodegenerative diseases. According to the number of the CAG repetitions in the gene coding huntingtin, the onset of the disease is in childhood (5%), in the middle age, which is the most common (90%) and in the older age (5%). Beginning of the disease is manifested by changes in behavior; including problems with coordination and movement. Later, there is a psychological change. The disease leads to death. Until now, there is no effective curative treatment. In 2009, we created a model of the transgenic minipigs (TgHD) carrying the N - terminal part of the human mutant huntingtin (mtHtt) at our Institute in Liběchov. The number of offsprings and the resemblance in physiology and morphology between the pig (Sus scrofa) and humans (Homo sapiens) give us opportunities not only to study changes not only in central nerve organs, but also in peripheral tissues. The reproductive problems of TgHD boars were observed as the first phenotypic changes. Therefore, my work focuses at first on a study of the reproduction parameters of TgHD boars as well as ultrastructural, immunocytochemical and biochemical changes in testes and spermatozoa. In PhD thesis, I described in details the reproductive defects in TgHD...
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Differential potential of neural crest cells
Wróblová, Aneta ; Krylov, Vladimír (advisor) ; Machoň, Ondřej (referee)
The neural crest is a transient structure formed during the neurulation which undergoes change of phenotype in the process of epithelial-mesenchymal transition. Subsequently, neural crest cells delaminate and migrate collectively and individually to their place of destination, where they differentiate into a broad repertoir of mesenchymal and non-mesenchymal cell types. This thesis aims to examine true differential potential of these cells based on crucial in vitro experiments. Neural crest cells show not only high migration potential, but also stem cells characteristics like multipotency and self-renewal capacity. I also provide answers to questions about cellular potency at the level of neural crest population and along anterior-posterior axis. It was necessary to clarify the essence of events leading to induction, specification, epithelial-mesenchymal transition and migration of neural crest cells, since they are crucial for their differentiation potential. A major role in differentiation as well as in previous processes plays the gene regulatory network which is comprised of mutually affecting signalling pathways. Studying the behaviour of migratory and post-migratory neural crest cells is important for the research of regenerative medicine and even cancer and neurocristopathic treatment.
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Quality evaluation of early human embryos regarding their developmental competence
Černíková, Terezie ; Krylov, Vladimír (advisor) ; Fulka, Josef (referee)
The mission of assisted reproductive medicine is to achieve the highest success rate of implantation and the subsequent birth of a healthy child. The current standard is the transfer of only one embryo and thus there is a great effort to develop an unbiased and accurate embryo assessment methods. This bachelor thesis is an overview of several selected techniques and their advantages and disadvantages. The selection of methods included in this thesis is based on their potential contribution to embryo assessment and the number and availability of relevant studies. I tried to compare the classical morphological approach with the newly developed methods monitoring vitality biomarkers, including genetic, metabolic and proteomic analysis. Because of the lack of information concerning techniques from clinical practice and large randomized trials, the comparison of new and old methods is questionable. Nevertheless, their principal differences can be compared. Conventional morphological assessment is based on the large extent of the subjective assessment by the assessor. On the other hand biomarker monitoring methods are based on empirical data. The outlook for the future is the complementarity of biomarker monitoring to morphological assessment. Key words: early embryo, human, developmental competence,...
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Molecular mechanisms of vertebrate limb regeneration
Onhajzer, Jakub ; Krylov, Vladimír (advisor) ; Soukup, Vladimír (referee)
Limb regeneration fascinates innumerable scientists for decades. Urodele amphibians can regenerate their limbs perfectly. This ability is preserved for a whole lifetime. However, they are not the only ones who regenerate their limbs. Second species are anuran amphibians, but their ability to promote limb regeneration take place only throughout a larval stage. Both groups belong to amphibians. Limbs are regeneated by the process called epimorphosis. The primary process is formation of blastema, mass of heterogeneous dedifferetiated cells, which are unipotent with the capacity to redifferentiate into only one cell type. Essential factor is the regulation of limb regeneration by numerous molecular mechanisms in order to achieve the perfect limb shape, without unwanted tumors. Mechanisms allowing limb regeneration in lower vertebrates would be applied via regenerative medicine in higher vertebrates in the future. Keywords: regeneration, limb, epimorphosis, dedifferentiation, vertebrates
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