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Michal Benc, Lazo Pendovski, Matej Murin, Frantisek Strejcek, Martin Morovic, Radek Prochazka and Jozef Laurincik

Abstract

The nucleolus is the cell organelle responsible for ribosome synthesis and, hence, for protein synthesis. In the mammalian oocyte, the nucleolus compacts into a dense sphere with no ribosome synthesis well in advance of ovulation. It seems, that this body is of utmost importance for the development of the embryo. It is unknown, however, how it exerts this essential function. During the last two decades, great attention has been paid to the study of nucleogenesis in oocytes and early embryos, with transcription of ribosomal DNA being evaluated as one of the criteria of normal development. In this review, we summarize some aspects of nucleolus transformation during oocyte growth, as well as during early embryonic development with possible impact on the quality of the embryos used in biomedical research. This knowledge in connection with further observations will substantially contribute to the development of new criteria suitable for evaluation of oocytes and embryos used in biomedical application.

Open access

Martin Morovic, Matej Murin, Frantisek Strejcek, Michal Benc, Dusan Paál, Olga Østrup, Heiner Niemann, Lazo Pendovski and Jozef Laurincik

Abstract

One of the main reason for the incorrect development of embryos derived from somatic cell nuclear transfer is caused by insufficient demethylation of injected somatic chromatin to a state comparable with an early embryonic nucleus. It is already known that the epigenetic enzymes transcription in oocytes and early embryos of several species including bovine and porcine zygotes is species-dependent process and the incomplete DNA methylation correlates with the nuclear transfer failure rate in mammals. In this study the transcription of DNA methyltransferase 1 and 3a (DNMT1, DNMT3a) genes in early embryonic stages of interspecies (bovine, porcine) nuclear transfer embryos (iSCNT) by RT-PCR were analyzed. Coming out from the diverse timing of embryonic genome activation (EGA) in porcine and bovine preimplantation embryos, the intense effect of ooplasm on transferred somatic cell nucleus was expected. In spite of the detection of ooplasmic DNA methyltransferases, the somatic genes for DNMT1 and DNMT3a enzymes were not expressed and the development of intergeneric embryos stopped at the 4-cell stage. Our results indicate that the epigenetic reprogramming during early mammalian development is strongly influenced by the ooplasmic environment.

Open access

Michal Benc, Frantisek Strejcek, Matej Murin, Martin Morovic, Stanislava Martinkova, Dominika Jettmarova, Lazo Pendovski, Josef Fulka and Jozef Laurincik

Abstract

An effort to improve development potential of early embryos is one of the main goals of biotechnology in the area of reproductive biology with application in veterinary or human medicine. Recent observations of the function of nucleolus or rather its forms before, during and after the fertilisation or parthenogenetic activation show the key role(s) of nucleolus in the processes of early genome activation. The nucleolus is a subnuclear structure (organelle) mainly involved in regulation of transcription and translation. This organelle has been characterized in detail by immunofluorescence, cell transfection and proteomics. This data was, however, mostly obtained in nucleoli of differentiated eukaryotic cells. Much less is known about the nucleolar structural changes and related functional processes in growing and fully grown mammalian oocytes, zygotes and early cleavage stage embryos, especially in the context of embryonic genome activation. It has been shown, that nucleoli in mammalian oocytes and early embryos have several forms and functions, which vary during the oocyte growth and embryonic development. Certain functions have not been fully described or explained, yet. The method of enucleolation, which allows to remove nucleoli from the oocytes or to exchange nucleoli between oocytes or zygotes, together with their proteomic and structural analyses brought new information about functions of nucleoli in oocytes and early cleavage-stage embryos and allowed to explain some new key roles of nucleoli during oocyte maturation and early embryonic development.