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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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Development and function of endocrine cells of the pancreas
Hamplová, Adéla ; Pavlínková, Gabriela (advisor) ; Berková, Zuzana (referee)
Diabetes mellitus affects nearly 300 million people in the world. The development of diabetes is caused by dysfunction or by reduction of insulin-producing β-cells that are part of the endocrine pancreas. Therefore, the most critical step for understanding the pathophysiology of diabetes and for restoring lost β cells is the identification of molecular cues that specify the cellular phenotype in the pancreas. This work is based on the hypothesis that the transcription factor NEUROD1 is a key factor for the development of the pancreas and for the maintenance of endocrine tissue function. Neurod1 conditional KO mutants (Neurod1CKO) were generated using the Cre-loxP system by crossing floxed Neurod1 mice with Isl1-Cre line. Immunohistochemical analyses of the pancreas at embryonic day 17.5 and postnatal day 0 showed that the deletion of Neurod1 negatively affected the development, organization of endocrine tissue, and total mass of pancreatic endocrine cells. To better understand molecular changes, quantitative PCR was used to analyse mRNA expression in the developing pancreas at the age of embryonic day 14.5 and postnatal day 1. Genes important for the development and function of the pancreas have been selected for the study of expression changes. These analyses showed changes in expression of genes...
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The legal status of the embryo from the perspective of European human rights protection
Havlíková, Barbora ; Scheu, Harald Christian (advisor) ; Svobodová, Magdaléna (referee)
Title The legal status of the embryo from the perspective of European human rights protection Abstract The thesis addresses the question regarding the legal status of the human embryo in European law. The aim of the thesis is to evaluate whether the embryo stands in the position of a legal object or legal subject and whether the legal status of embryo is in compliance with natural law. For this purpose the thesis firstly defines the status in the field of natural law, i.e. the moral status of embryo. Afterwards, the thesis examines the legal position of embryo in human rights protection. The thesis answers the question whether the embryo is subject of human rights documents. The legal status of embryo is analyzed in relation to the subjectivity of human rights in general as well as in relation to specific rights. The interpretation of legal terms "human being" and "everyone" contribute to a better understanding of legal status of embryo in general. The analysis of the specific rights, namely the right to life, human dignity, personal integrity and non-discrimination, shows the complexity of the potential legal subjectivity of embryo in the area of human rights. The evaluation of the legal status of human embryo in the area of human rights protection is complemented by an assessment of legal status in other...
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Tripolar cell cleavage - morphokinetic evaluation of the human embryo development by the time-lapse system
Jandová, Oldřiška ; Hlinka, Daniel (advisor) ; Anger, Martin (referee)
The principle of time-lapse system is based on the continuous sensing of the development of the pre-implantation embryo and the assessment of its morphology. This is advantageous in assessing the quality and implantation potential of embryos, which are subsequently evaluated according to certain established criteria. This gives the opportunity to select the highest quality embryo to be implanted into the mother's womb during embryo transfer. This system also allows us to detect any abnormalities in embryo development. This is very important, because the occurrence of abnormalities in early embryonic cell division is quite common. Morphological evaluation of embryos indicates a high incidence of tripolar mitosis during this early embryo development. The result of this division is three blastomers instead of two, which is associated with an irregular chromosome separation, each of them may contain a different number of chromosomes. In the case where conventional embryo observation is used to observe embryo morphology at longer intervals, it may be that the changes associated with tripolar mitosis are not detected at all, and such embryos appear to be falsely prosperous and are often selected for transfer. This can have serious consequences in the case of implantation, because these embryos are not...
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The view of statut human embryo from ethical point of view
BŘEZINOVÁ, Nicola
The work deals with looking at the status of the human embryo. The structure of the work is divided into several chapters that deal with a particular topic. The first chapter deals with the historical view of the adoption of a human embryo, in the second chapter is devoted to philosophical adopting the human embryo and view individual philosophers on this issue. The third chapter is devoted to the ethical acceptance of the issue. Here is discussed an ethical perspective on human embryo status and access two main streams which occur in this area. The next part is devoted to the present, both in terms of the ethical-philosophical, and in terms of health. In the medical section includes prenatal communication between mother and child, and research on embryonic stem cells, which is associated with a number of ethical issues.
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Legal and ethical status of the human embryo through the eyes of the young generation
WÁGNEROVÁ, Lucie
There has been the question of the approach to the unborn human foetus which has been dealt with for many years. The moral, ethic or legal questions appear during the discussions. In this context we could find many different opinions related to the upbringing, religion or morality. It is hard to find any general agreement. Nowadays there are new questions connected to the intervention of reproduction, the research of embryo cells or the issues of abortion, which is perceived as the women's right. There are about 44 billions of abortions these days. According to these problems there is the need of the social discussion about the understanding of embryo and when we can consider it as a human being. The theoretical part of my bachelor thesis deals with the legal point of view to the role of human embryo and its securing in our legal system. The Charter of Fundamental Rights and Freedoms, which is a part of the Constitution of the Czech Republic, determines that everybody has the right for living as well as the human life should be protected before the delivery. There is not and there also cannot be determined clearly when the human life begins. The answer for this question is connected to the moral and ethic principles. My thesis deals with the different points of view to this question, if the fertilized ovum is or is not the human. There are also many various arguments about when we can consider the genesis of human life. The answer for these questions is not clear and we cannot find out the definite concordance. The other part of my thesis deals with ethic dilemmas connected to the abortion and the intervention to the reproduction as well as the problems of cloning. The practical part of my bachelor's thesis deals with the legal and ethic status of embryo mainly to the questions of the genesis of human life and the problems of the abortion. This thesis do not provide the clear answer to the question since when we can date the genesis of the life but it represents the opinions of the young generation (18 - 30 years old). The main aim of the research was to find out the point of view of the young people in the Czech Republic. There were questions about the reaction to the unborn child, the genesis of human life, who decides about the life of the unborn child and the abortion. I compare the questions according to the sex, ages, religion and parenthood. The practical part was made by the quantitative research questionnaires. It consists of 250 respondents who were asked by the on-line or printed questionnaire. It was chosen the young generation between 18 - 30 years old. The questionnaire contains seven fact-finding questions and four distinctive ones. It was worked with the data which were acquired from the questionnaire. The first hypothesis, 50% of respondents believe that the human life begins with the conception, was not confirmed. There were only 30% of respondents who believe that question. The second hypothesis, the approval of abortion, was confirmed because there were 90% respondents who agreed. The third hypothesis could not be confirmed. In the discussion the results were compared with the opinions of the authors from the theoretical part of my thesis. The end of my thesis deals with the general results. We cannot find the right answer to the question of the beginning of human life. Every person has a subjective point of view. We should realize every possible risks and think about who has the right to intervene to the substance of human life and how far can technology goes. This bachelor's thesis do not have the clear opinion but it provides the facts and the suggestion for thinking for creating its own attitude to these questions.
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Regulation of development of mouse parthenogenetic embryos
Jettmarová, Dominika ; Fulka, Josef (advisor) ; Kaňka, Jiří (referee)
The development of mouse (Mus musculus) haploid parthenogenetic embryos does not reach the same level as normal embryos. The aim of this diploma thesis was to find out whether haploid parthenogenetic embryos of mice differ in the nucleocytoplasmic ratio. The volume of the nucleus increases with ploidity. The nucleocytoplasmic ratios of haploid embryos do not significantly change between the two-cell and four-cell stage (p = 0.052), there is a significant difference (p < 0.001) for diploid and tetraploid embryos. Non-standard nucleocytoplasmic ratio could be related to the problematic development. Understanding the regulation of preimplantational development of parthenogenetic embryos will increase the efficiency of haploid embryonic stem cell derivation.
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Effect of incubation temperature on growth of the working and conducting myocardium in the embryonic chick
Skuhrová, Kristýna ; Sedmera, David (advisor) ; Gregorovičová, Martina (referee)
It was shown almost 50 years ago that hypothermic incubation of chicken embryos results in a reduction in the size of embryos and an increase in the heart weight, presumably by hypertrophy (increase in cell volume). The chicken embryos were incubated in normothermia (37.5 ř C) and hypothermia (33.5 ř C) from the eleventh embryonic day. On the 17th day, the embryos were weighed and then their hearts were weighed. In agreement with the previous results, hypothermic embryos were 29% smaller and their hearts 18% heavier. The heart-to-body weight ratio was 67% higher in the hypothermic group. The measured cell size was very similar in the target areas and it was also between the two groups. The left ventricle width was twofold that the right one and the difference was not significantly higher in the hypothermia model. Purkinje fibers, the terminal part of the conduction system, were smaller than the working cardiomyocytes. Purkinje fibers were slightly enlarged after hypothermic incubation. The proliferation rate was measured by immunohistochemical labeling of anti-phospho histone H3. The experimental group showed much higher proliferation rate; it reached statistical significance in the right ventricle. Thus, hypothermic incubation resulted in increased growth of embryonic heart based on hyperplasia...
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Legal aspects of stem cells research
Česáková, Barbora ; Šustek, Petr (advisor) ; Salač, Josef (referee)
This thesis deals with the issue of stem cell research and its legal aspects. The issue raises many ethical questions, especially due to its closed connection to the topics of embryonic research and right to life. In the first introductory chapter the thesis explains the topic of stem cells from biological point of view and defines and classifies stem cells by their origin and differential potential. In the second chapter the thesis briefly summarizes the history of stem cell research and deals with the ethical questions raised by closed connection of the topics of embryonic stem cell research and in vitro fertilization to the right to life. The second chapter also notes some recent possibilities and medical methods connected with stem cells, such as gene therapy and diagnostics and new eugenics. In the following chapters the thesis concentrates on the legal aspects of the topic on both national and international level. Specifically, the third chapter puts the issue of stem cells research to the contexts of the two most important conventions - the Convention for the Protection of Human Rights and Fundamental Freedoms and the Convention on Human Rights and Biomedicine. The fourth chapter concentrates on legislation of Czech Republic regulating the legal status of embryo and its protection in the...
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Mode of reproduction and reproductive assurance of diploid and polyploid hawkweeds (Hieracium s. str.)
Zdvořák, Pavel ; Mráz, Patrik (advisor) ; Krahulcová, Anna (referee)
The mode of reproduction can greatly influence the demography and the evolutionary success of the taxon. In the case of autonomous asexual formation seeds are apomictic taxa fully independent of pollinators and compatible partners. For sexual taxa with strict autoincompatibility it is the opposite, i.e. sexual taxa need pollinators and compatible partners for birth of offspring. Therefore, in marginal population and for more extreme situation with lower pollinating activity will have apomictic taxa a higher level of reproductive assurance than sexual taxa vascular plants. This hypothesis was tested in natural populations of apomictic and sexual taxa. In the diploma thesis we therefore investigate the method mode of reproduction and reproductive assurance of 52 taxa of the genus Hieracium s. str. (family Asteraceae) in Europe. Of these, 12 were diploid sexually diploid taxa and 42 polyploid apomictic reproductive taxa. From these taxa we harvested seeds from fully developed capitulum and we determined the potential (total number of seeds in the capitulum) and the realized (the percentage of well-developed seeds at the capitulum). The ploidy of the offspring (the embryos and the seedling) and method origins of seeds we examined using flow cytometry. The results show that the plants of diploid species...
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