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Mouse oocytes nucleoli rescue embryonic development of porcine enucleolated oocytes

Published online by Cambridge University Press:  20 November 2017

Martin Morovic*
Affiliation:
Department of Zoology and Anthropology, Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nabrezie mladeze 91, 949 74, Nitra, Slovakia.
Frantisek Strejcek
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic.
Shoma Nakagawa
Affiliation:
Institute of Animal Science, Prague, Czech Republic. Centre de Recherche du Centre Hospitalier du l'Université de Montreal (CRCHUM), Quebec, Canada.
Rahul S. Deshmukh
Affiliation:
Bombay Veterinary College, Mumbai, India.
Matej Murin
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic.
Michal Benc
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic. Institute of Animal Science, Prague, Czech Republic.
Helena Fulka
Affiliation:
Institute of Animal Science, Prague, Czech Republic. Institute of Molecular Genetics of the ASCR, Prague, Czech Republic.
Hirohisa Kyogoku
Affiliation:
RIKEN Kobe, Kobe, Japan.
Lazo Pendovski
Affiliation:
Ss. Cyril and Methodius University in Skopje, Republic of Macedonia.
Josef Fulka Jr
Affiliation:
Institute of Animal Science, Prague, Czech Republic.
Jozef Laurincik
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic. Institute of Animal Physiology and Genetics ASCR, Libechov, Czech Republic. Catholic University, Ruzomberok, Slovakia.
*
All correspondence to: Martin Morovic. Department of Zoology and Anthropology, Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nabrezie mladeze 91, 949 74, Nitra, Slovakia. Tel: +421 37 6408 712. Fax: +421 37 6408 556. E-mail: [email protected]

Summary

It is well known that nucleoli of fully grown mammalian oocytes are indispensable for embryonic development. Therefore, the embryos originated from previously enucleolated (ENL) oocytes undergo only one or two cleavages and then their development ceases. In our study the interspecies (mouse/pig) nucleolus transferred embryos (NuTE) were produced and their embryonic development was analyzed by autoradiography, transmission electron microscopy (TEM) and immunofluorescence (C23 and upstream binding factor (UBF)). Our results show that the re-injection of isolated oocyte nucleoli, either from the pig (P + P) or mouse (P + M), into previously enucleolated and subsequently matured porcine oocytes rescues their development after parthenogenetic activation and some of these develop up to the blastocyst stage (P + P, 11.8%; P + M, 13.5%). In nucleolus re-injected 8-cell and blastocyst stage embryos the number of nucleoli labeled with C23 in P + P and P + M groups was lower than in control (non-manipulated) group. UBF was localized in small foci within the nucleoli of blastocysts in control and P + P embryos, however, in P + M embryos the labeling was evenly distributed in the nucleoplasm. The TEM and autoradiographic evaluations showed the formation of functional nucleoli and de novo rRNA synthesis at the 8-cell stage in both, control and P + P group. In the P + M group the formation of comparable nucleoli was delayed. In conclusion, our results indicate that the mouse nucleolus can rescue embryonic development of enucleolated porcine oocytes, but the localization of selected nucleolar proteins, the timing of transcription activation and the formation of the functional nucleoli in NuTE compared with control group show evident aberrations.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2017 

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