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Influence of lamin A/C on paternal pronucleus DNA demethylation
Václavková, Vendula ; Fulková, Helena (advisor) ; Žatecká, Eva (referee)
A series of dynamic epigenetic changes need to happen to rebuild the gamete's genome after fertilization and to secure the totipotency of the zygote. The processes can be observed in both parental pronuclei, although the machinery providing the epigenetic dynamics probably lies in the maternal oocyte, more specifically in the germinal vesicle (GV). GV can be divided into two fractions: a soluble and an insoluble fraction. The probable ability of the soluble fraction is remodelling the sperm head and the paternal pronucleus formation, while the insoluble fraction could be involved in reprogramming of the paternal genome and ensuring the developmental competence. This thesis was focused on one of the epigenetic regulations which was DNA methylation and its rapid loss after fertilization. Putative mechanisms, which can occur there are the oxidation of methylated bases and their subsequent repair and replacement through the base excision repair (BER). This thesis tested a hypothesis that a protein lamin A/C, found in the insoluble fraction of the GV, is a possible candidate participating in the active demethylation through BER. To examine the role of lamin A/C in BER, it was required to prepare such GV oocytes, which would contain both the soluble part of GV and protein lamin A / C. The selective...
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