Maintenance of genome stability is essential for every living cell as genetic information is repeatedly challenged during DNA replication in each cell division event. Errors, defects, delays, and mistakes that arise during mitosis or meiosis lead to an activation of DNA repair processes and in case of their failure, programmed cell death, i.e. apoptosis, could be initiated. Fam208a is a protein whose importance in heterochromatin maintenance has been described recently. In this work, we describe the crucial role of Fam208a in sustaining the genome stability during the cellular division. The targeted depletion of Fam208a in mice using CRISPR/Cas9 leads to embryonic lethality before E12.5. We also used the siRNA approach to downregulate Fam208a in zygotes to avoid the influence of maternal RNA in the early stages of development. This early downregulation increased arresting of the embryonal development at the two-cell stage and occurrence of multipolar spindles formation. To investigate this further, we used the yeast two-hybrid (Y2H) system and identified new putative interaction partners Gpsm2, Amn1, Eml1, Svil, and Itgb3bp. Their co-expression with Fam208a was assessed by qRT-PCR profiling and in situ hybridisation [1] in multiple murine tissues. Based on these results we proposed that Fam208a functions within the HUSH complex by interaction with Mphosph8 as these proteins are not only able to physically interact but also co-localise. We are bringing new evidence that Fam208a is multi-interacting protein affecting genome stability on the level of cell division at the earliest stages of development and also by interaction with methylation complex in adult tissues. In addition to its epigenetic functions, Fam208a appears to have an additional role in zygotic division, possibly via interaction with newly identified putative partners Gpsm2, Amn1, Eml1, Svil, and Itgb3bp.

• MeSH
• aparát dělícího vřeténka metabolismus   MeSH
• buněčné dělení genetika   fyziologie   MeSH
• CRISPR-Cas systémy MeSH
• embryonální vývoj genetika   fyziologie   MeSH
• fosfoproteiny metabolismus   MeSH
• HEK293 buňky MeSH
• jaderné proteiny fyziologie   MeSH
• letální geny MeSH
• lidé MeSH
• malá interferující RNA genetika   farmakologie   MeSH
• multiproteinové komplexy MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• nestabilita genomu MeSH
• RNA interference MeSH
• vývojová regulace genové exprese * MeSH
• zvířata MeSH
• zygota metabolismus   MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• embryo savčí MeSH
• experimenty na zvířatech MeSH
• myši MeSH
• ovum růst a vývoj   ultrastruktura   MeSH
• zygota MeSH
• Check Tag
• myši MeSH

Aneuploidy is the most frequent single cause leading into the termination of early development in human and animal reproduction. Although the mouse is frequently used as a model organism for studying the aneuploidy, we have only incomplete information about the frequency of numerical chromosomal aberrations throughout development, usually limited to a particular stage or assumed from the occurrence of micronuclei. In our study, we systematically scored aneuploidy in in vivo mouse embryos, from zygotes up to 16-cell stage, using kinetochore counting assay. We show here that the frequency of aneuploidy per blastomere remains relatively similar from zygotes until 8-cell embryos and then increases in 16-cell embryos. Due to the accumulation of blastomeres, aneuploidy per embryo increases gradually during this developmental period. Our data also revealed that the aneuploidy from zygotes and 2-cell embryos does not propagate further into later developmental stages, suggesting that embryos suffering from aneuploidy are eliminated at this stage. Experiments with reconstituted live embryos revealed, that hyperploid blastomeres survive early development, although they exhibit slower cell cycle progression and suffer frequently from DNA fragmentation and cell cycle arrest.

• MeSH
• aneuploidie * MeSH
• blastomery cytologie   metabolismus   MeSH
• buněčný cyklus MeSH
• embryo savčí cytologie   metabolismus   MeSH
• embryonální vývoj * MeSH
• fertilizace in vitro MeSH
• myši MeSH
• těhotenství MeSH
• zvířata MeSH
• zygota cytologie   metabolismus   MeSH
• Check Tag
• myši MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Dietary phospholipids (PLs) and their derivatives have proved active in suppression of various health problems and conditions including cancer. In this work we compared the effect of dietary phospholipids from hen egg yolk enriched with N-acyl ether-phosphatidyl ethanolamine (NAEPE) termed bioactive phospholipids (BAP+ preparation) with PLs lacking NAEPE (BAP- preparation) on the growth of transformed cells in vitro and on the promotion and progression of experimental tumours in vivo. For the in vivo experiments we used the chicken model in which liver, lung, and kidney tumours arose via natural selection from single cells initiated by experimentally introduced somatic mutations caused by insertional mutagenesis. Mutagenized animals were fed BAP+ or BAP- diet in various regimens. We observed that BAP+ at low concentrations killed cells of various tumour cell lines in culture but did not compromise viability of non-transformed cells. Oral administration of the BAP+ preparation efficiently reduced progression of all tumour types. However, it did not significantly reduce the number of already initiated tumours and their growth when BAP+ was discontinued. Our data suggest that NAEPE combined with hen egg PLs significantly interferes with tumour progression, possibly through the inhibition of tumour cell viability.

• MeSH
• aplikace orální MeSH
• ethanolaminy chemie   farmakologie   MeSH
• fosfolipidy aplikace a dávkování   chemie   farmakologie   MeSH
• kultivované buňky MeSH
• kuřecí embryo MeSH
• modely nemocí na zvířatech MeSH
• nádorové buněčné linie MeSH
• nádory farmakoterapie   patofyziologie   MeSH
• proliferace buněk účinky léků   MeSH
• vaječný žloutek chemie   MeSH
• viabilita buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• kuřecí embryo MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• buněčná diferenciace MeSH
• mezibuněčné spoje MeSH
• ovariální folikul cytologie   MeSH
• ovulace MeSH
• ovum cytologie   MeSH

• MeSH
• buněčná diferenciace MeSH
• buněčná membrána fyziologie   MeSH
• embryo savčí fyziologie   MeSH
• geny MeSH
• lidé MeSH
• ovum fyziologie   MeSH
• těhotenství MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH

Early embryonic development is characterized by a plethora of very complex and simultaneously operating processes, which are constantly changing cellular morphology and behaviour. After fertilization, blastomeres of the newly created embryo undergo global epigenetic changes and simultaneously initiate transcription from the zygotic genome and differentiation forming separate cell lineages. Some of these mechanisms were extensively studied during the last several decades and valuable insight was gained into how these processes are regulated at the molecular level. We have, however, a still very limited understanding of how multiple events are coordinated during rapid development of an early mammalian embryo. In this review, we discuss some aspects of early embryonic development in mammals, namely the fidelity of chromosome segregation and occurrence of aneuploidy, as well as the clinical applications of cell cycle monitoring in human embryos.

• MeSH
• aneuploidie MeSH
• aparát dělícího vřeténka metabolismus   MeSH
• blastomery metabolismus   MeSH
• buněčný cyklus genetika   MeSH
• embryo savčí cytologie   metabolismus   MeSH
• embryonální vývoj genetika   MeSH
• lidé MeSH
• segregace chromozomů genetika   MeSH
• těhotenství MeSH
• zvířata MeSH
• zygota cytologie   metabolismus   MeSH
• Check Tag
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

• MeSH
• embryo savčí genetika   MeSH
• embryonální vývoj MeSH
• finanční podpora výzkumu jako téma MeSH
• fúze buněk MeSH
• myši MeSH
• oocyty MeSH
• polyploidie MeSH
• zvířata MeSH
• zygota cytologie   genetika   MeSH
• Check Tag
• myši MeSH
• zvířata MeSH

Maintenance of genome stability is essential for every living cell as genetic information is repeatedly challenged during DNA replication in each cell division event. Errors, defects, delays, and mistakes that arise during mitosis or meiosis lead to an activation of DNA repair processes and in case of their failure, programmed cell death, i.e. apoptosis, could be initiated. Fam208a is a protein whose importance in heterochromatin maintenance has been described recently. In this work, we describe the crucial role of Fam208a in sustaining genome stability during cellular division. The targeted depletion of Fam208a in mice using CRISPR/Cas9 led to embryonic lethality before E12.5. We also used the siRNA approach to downregulate Fam208a in zygotes to avoid the influence of maternal RNA in the early stages of development. This early downregulation increased arresting of the embryonal development at the two-cell stage and the occurrence of multipolar spindles formation. To investigate this further, we used the yeast two-hybrid (Y2H) system and identified new putative interaction partners Gpsm2, Svil, and Itgb3bp. Their co-expression with Fam208a was assessed by RT-qPCR profiling and in situ hybridization [1] in multiple murine tissues. Based on these results we proposed that Fam208a functions within the HUSH complex by interaction with Mphosph8 as these proteins are not only able to physically interact but also co-localise. We are bringing new evidence that Fam208a is a multi-interacting protein affecting genome stability on the cell division level at the earliest stages of development and by interaction with methylation complex in adult tissues. In addition to its epigenetic functions, Fam208a appears to have an important role in the zygotic division, possibly via interaction with newly identified putative partners Gpsm2, Svil, and Itgb3bp.

• MeSH
• CRISPR-Cas systémy MeSH
• embryonální vývoj * MeSH
• epigeneze genetická * MeSH
• fosfoproteiny genetika   metabolismus   MeSH
• jaderné proteiny antagonisté a inhibitory   fyziologie   MeSH
• metylace DNA MeSH
• mitóza * MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• nestabilita genomu * MeSH
• těhotenství MeSH
• zvířata MeSH
• zygota fyziologie   MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• biologie buňky MeSH
• molekulární biologie MeSH
• Publikační typ
• monografie MeSH

• Konspekt
• Biochemie. Molekulární biologie. Biofyzika
• NLK Obory
• biologie
• cytologie, klinická cytologie