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Effect of pergafast 201 on selected markers of in vitro maturation of porcine oocytes.
Procházková, Bára ; Petr, Jaroslav (advisor) ; Benc, Michal (referee)
Pergafast 201 (N-(p-Toluenesulfonyl)-N'-(3-p-toluenesulfonyloxyphenyl)urea) is a patented color developer that is offered as an alternative to the widely used bisphenols A and S (BPA, BPS). These substances have been shown to have a negative effect on human health and reproduction even in very low doses, and they are endocrine disruptors affecting, among other things, the process of meiotic maturation. The effects of pergafast 201 on the mammalian organism during oogenesis have not yet been monitored. The aim of this work was to evaluate the effect of pergafast 201 on selected specific markers of meiotic maturation after exposure of porcine oocytes to pergafast 201 in low concentrations in vitro. This work is the first to describe the negative effects of pergafast 201 on the maturation of porcine oocytes in vitro, confirming the negative effect of pergafast on the course of meiotic maturation and on the ability of the oocyte to reach the final stage of maturation, as well as its effect on the increased incidence of abnormal dividing spindles. The data show the effect of PF201 on the methylation of epigenetic markers H3K4me2 and H3K9me3. Preliminary results also indicate its effect on the mRNA expression of estrogen receptors α and β. Collectively, these results indicate the possible risks...
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Effect of on in vitro maturation of porcine oocytes
Laštůvková, Viktorie ; Petr, Jaroslav (advisor) ; Benc, Michal (referee)
Lysmeral (Lilial, Butylphenyl Methylpropional) is a synthetically produced aldehyde. It is used for its typical lily of the valley-like scent as a perfume ingredient in cosmetic products. It is typically found, for example, in cleaning products. It is on the list of the 26 most well-known synthetic allergens and has been shown to be present in human urine and subsequently in wastewater. According to some opinions, lysmeral should be classified as an endocrine disruptor, which are substances that can interfere with the body's endocrine system. As these substances often affect mammalian reproduction, it is in society's interest to address them, given that they are commonly found in the environment. Lysmeral was not properly tested before being placed on the market and all its effects on organisms are still unknown. The hypothesis of this thesis is that exposure to lysmeral causes undesirable changes in the meiotic maturation of porcine oocytes in vitro and the aim is to investigate the effect of lysmeral, at concentrations corresponding to normal human exposures, on specific markers of meiotic maturation of porcine oocytes. The results confirm the negative effect of lysmeral on the smooth progression of meiotic maturation and the achievement of the final stage of porcine oocyte maturation in vitro....
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Use of interspecific ICSI method for assessing the quality of the retained biological material from genetic resources
Vacková, Kristýna ; Benc, Michal (advisor) ; Barnetová, Irena (referee)
Intracytoplasmic sperm injection, or ICSI, is a commonly used method in assisted reproduction centers and is also an effective tool for a fertilization research. The interspecific ICSI is the suitable method for production of zygotes, which are formed by association of gametes of different, distantly unrelated species. It can bypass natural interspecies barriers and create so-called interspecific zygotes. This can be used to evaluation properties of genetic material. In this work, this method was used for evaluation of the quality of stored sperm originating from male goats of the white shorthair goat genetic resource. The aim of this work was to evaluate the effect of cryopreservation on the quality of goat sperm, which was frozen in three different ways at two different breeding stations. Freshly collected semen served as a control group. The heads of xenogeneic goat sperm were injected into mature mouse oocytes. The injected oocytes were allowed to develop to the zygote stage, in which both female (mouse) and male (goat) pronuclei was formed. In male pronuclei, DNA damage was assessed by immunofluorescence staining with anti-γH2AX antibody, a marker of DNA breaks. The results obtained by measuring the level of γH2AX fluorescence intensity in the paternal pronuclei show that the values differ...
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