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Male fertility disorders and sperm damage related to COVID-19 disease.
Pikulová, Simona ; Komrsková, Kateřina (advisor) ; Ješeta, Michal (referee)
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of the coronavirus disease 2019 (COVID-19), which has spread rapidly worldwide since the end of 2019. The virus causes serious health problems and damage to human organs, especially the lungs, kidneys, brain, heart and testicles, which can be fatal. This work deals with the entry of SARS- CoV-2 virus into the testicles and sperm damage due to infection. Receptors are described in more detail, specifically the membrane receptor ACE2, which mediates the entry of the virus into germ cells (spermatogonia) and somatic testicular cells (Sertoli and Leydig), which express this receptor. In this work, the mechanisms causing infection are described and attention is focused to changes in the signaling pathways due to SARS-CoV-2 and ACE2 binding. Consequently, the presence of the virus leads to hormonal imbalances, local oxidative stress and inflammation, which affect the course of spermatogenesis and can cause pathological conditions of spermatozoa such as motility disorders or DNA fragmentation and thus adversely affect male fertility. Key words: COVID-19, SARS-CoV-2, male infertility, sperm, ACE2
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Identifikace genů nezbytně nutných pro normální průběh oplození a preimplantačního vývoje skotu v podmínkách in vitro
Toralová, Tereza ; Kaňka, Jiří (advisor) ; Pěknicová, Jana (referee) ; Ješeta, Michal (referee)
The aim of this work was to find and characterize genes that seem to be important for normal preimplantation development. We characterized three genes in more detail - centromeric protein F (CENPF; mitosin), ubiquitin C-terminal hydrolase-L1 (UCHL1) and nucleophosmin (NPM1; B23; numatrin; NO38). CENPF and nucleophosmin were shown to start their expression at late 8-cell stage, i.e. at major embryonic genome activation (EGA), and were hence supposed to be important during bovine preimplantation development. CENPF plays crucial role during cell division, especially by mediating the interaction of kinetochores and microtubules. Nucleophosmin is a multifunctional nucleolar phosphoprotein, whose most important roles are rRNA processing, chaperoning, ribosome biogenesis and centriole duplication control. Further, we characterized the role of UCHL1 during fertilization of bovine oocytes. UCHL1 is a deubiquitinating enzyme that controls cytoplasmic protein degradation, recycling of free ubiquitin from proteasome products and is involved in regulation of physiological apoptosis. We studied the function of CENPF and nucleophosmin using RNA interference approach. Since UCHL1 protein is very stable, this method is not suitable for studying the UCHL1 function. We thus used two UCHL1 inhibitors that block its...
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Effect of hydrogen sulphide on aging of porcine oocytes in vitro
Krejčová, Tereza ; Petr, Jaroslav (advisor) ; Ješeta, Michal (referee)
Unfertilized or parthenogenetically non-activated porcine oocytes matured in vitro conditions are subjected to a process known as aging. During such development, porcine oocytes undergo the complex of the structural and functional changes, which can result in spontaneous parthenogenetic activation, fragmentation or lysis. After three days of culture in our condition, 23% of oocytes remained at the stage of metaphase II, 48% of oocytes were spontaneously parthenogenetically activated, 26% of oocytes were subjected to fragmentation and 3% of oocytes were lysed. The complete suppression of porcine oocyte fragmentation during the process of aging occurred during oocytes cultured in medium with sulphide donor Na2S in concentrations 150 µM and 300 µM. Inhibition of enzymes catalyzing the synthesis of hydrogen sulfide in the oocytes during the process of aging (cystathionine-gamma-lyase and cystathionine beta-synthase), iniciates earlier onset of oocytes fragmentation. The effect of both inhibitors could be completely reversed by using sulphide donor Na2S. The process of aging in porcine oocytes significantly reduces the success of the activation processes. Parthenogenetic activation occurred in 94% of pig oocytes, which were not subjected to aging. The proportion of activated oocytes after exposure to 24...
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