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
- Spindle Apparatus metabolism MeSH
- Cell Division genetics physiology MeSH
- CRISPR-Cas Systems MeSH
- Embryonic Development genetics physiology MeSH
- Phosphoproteins metabolism MeSH
- HEK293 Cells MeSH
- Nuclear Proteins physiology MeSH
- Genes, Lethal MeSH
- Humans MeSH
- RNA, Small Interfering genetics pharmacology MeSH
- Multiprotein Complexes MeSH
- Mice, Inbred C57BL MeSH
- Mice, Knockout MeSH
- Mice MeSH
- Genomic Instability MeSH
- RNA Interference MeSH
- Gene Expression Regulation, Developmental * MeSH
- Animals MeSH
- Zygote metabolism MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't 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
- Aneuploidy * MeSH
- Blastomeres cytology metabolism MeSH
- Cell Cycle MeSH
- Embryo, Mammalian cytology metabolism MeSH
- Embryonic Development * MeSH
- Fertilization in Vitro MeSH
- Mice MeSH
- Pregnancy MeSH
- Animals MeSH
- Zygote cytology metabolism MeSH
- Check Tag
- Mice MeSH
- Pregnancy MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't 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
- Administration, Oral MeSH
- Ethanolamines chemistry pharmacology MeSH
- Phospholipids administration & dosage chemistry pharmacology MeSH
- Cells, Cultured MeSH
- Chick Embryo MeSH
- Disease Models, Animal MeSH
- Cell Line, Tumor MeSH
- Neoplasms drug therapy physiopathology MeSH
- Cell Proliferation drug effects MeSH
- Egg Yolk chemistry MeSH
- Cell Survival drug effects MeSH
- Animals MeSH
- Check Tag
- Chick Embryo MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't 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
- Aneuploidy MeSH
- Spindle Apparatus metabolism MeSH
- Blastomeres metabolism MeSH
- Cell Cycle genetics MeSH
- Embryo, Mammalian cytology metabolism MeSH
- Embryonic Development genetics MeSH
- Humans MeSH
- Chromosome Segregation genetics MeSH
- Pregnancy MeSH
- Animals MeSH
- Zygote cytology metabolism MeSH
- Check Tag
- Humans MeSH
- Pregnancy MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review 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 Systems MeSH
- Embryonic Development * MeSH
- Epigenesis, Genetic * MeSH
- Phosphoproteins genetics metabolism MeSH
- Nuclear Proteins antagonists & inhibitors physiology MeSH
- DNA Methylation MeSH
- Mitosis * MeSH
- Mice, Inbred C57BL MeSH
- Mice, Knockout MeSH
- Mice MeSH
- Genomic Instability * MeSH
- Pregnancy MeSH
- Animals MeSH
- Zygote physiology MeSH
- Check Tag
- Male MeSH
- Mice MeSH
- Pregnancy MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
... Underlie Evolution 6 -- Even Single Cells Can Have Sex 7 -- We Develop from a Single Cell 8 -- Stem ... ... Growth and Development -- 20 REGULATING THE EUKARYOTIC -- CELL CYCLE 847 -- 20.1 -- Overview of the ... ... Murder by Activating Caspases 943 -- 22 THE MOLECULAR CELL BIOLOGY -- OF DEVELOPMENT 949 -- 22.1 -- ... ... Highlights of Development -- 950 -- Development Progresses from Egg and Sperm to an Early Embryo 950 ... ... -- As the Embryo Develops, Cell Layers Become Tissues and Organs 951 -- Genes That Regulate Development ...
6th ed. xxxvii, 1150 s. : il., tab. ; 29 cm
- MeSH
- Cell Biology MeSH
- Molecular Biology MeSH
- Publication type
- Monograph MeSH
- Conspectus
- Biochemie. Molekulární biologie. Biofyzika
- NML Fields
- biologie
- cytologie, klinická cytologie
