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Intranuclear characteristics of pig oocytes stained with brilliant cresyl blue and nucleologenesis of resulting embryos

Published online by Cambridge University Press:  09 August 2019

Matej Murin ,
Frantisek Strejcek ,
Alexandra Bartkova ,
Martin Morovic Open the ORCID record for Martin Morovic [Opens in a new window] ,
Michal Benc ,
Radek Prochazka ,
Andrea Lucas-Hahn ,
Lazo Pendovski  and
Jozef Laurincik
Matej Murin*
Affiliation:
Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Rumburska 89, 277 21 Libechov, Czech Republic
Frantisek Strejcek
Affiliation:
Constantine the Philosopher University in Nitra, Faculty of Natural Sciences, Nabrezie mladeze 91, 94974 Nitra, Slovak Republic
Alexandra Bartkova
Affiliation:
Constantine the Philosopher University in Nitra, Faculty of Natural Sciences, Nabrezie mladeze 91, 94974 Nitra, Slovak Republic
Martin Morovic
Affiliation:
Constantine the Philosopher University in Nitra, Faculty of Natural Sciences, Nabrezie mladeze 91, 94974 Nitra, Slovak Republic
Michal Benc
Affiliation:
Institute of Animal Science, Pratelstvi 815, 10400 Prague Uhrineves, Czech Republic Constantine the Philosopher University in Nitra, Faculty of Natural Sciences, Nabrezie mladeze 91, 94974 Nitra, Slovak Republic
Radek Prochazka
Affiliation:
Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Rumburska 89, 277 21 Libechov, Czech Republic
Andrea Lucas-Hahn
Affiliation:
Friedrich Loeffler Institute, Institute of Farm Animal Genetics, Holtystrasse 10, D-31535 Neustadt, Germany
Lazo Pendovski
Affiliation:
Ss. Cyril and Methodius University in Skopje, Republic of Macedonia.
Jozef Laurincik
Affiliation:
Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Rumburska 89, 277 21 Libechov, Czech Republic Constantine the Philosopher University in Nitra, Faculty of Natural Sciences, Nabrezie mladeze 91, 94974 Nitra, Slovak Republic
*
Address for correspondence: Matej Murin. Institute of Animal Physiology and Genetics CAS, v. v. i., Rumburska 89, 277 21 Libechov, Czech Republic. Tel: +420 315 639 544. E-mail: [email protected]
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Summary

Brilliant cresyl blue (BCB) vital labelling is a powerful method for analyzing the quality of porcine cumulus–oocyte complexes. Our aim was to investigate the correlation between the selection of porcine oocytes using BCB labelling and selected intranuclear characteristics of porcine oocytes and parthenotes. Moreover, BCB labelling was correlated with the diameter of the oocyte and the developmental potential of the parthenotes. The following methods were used: BCB labelling, measurement of the diameter of the oocyte, parthenogenetic activation, immunocytochemistry, transmission electron microscopy, enucleation and relative protein concentration (RPC) analysis. We determined that the diameter of the oocytes in the BCB-positive (BCB+) group was significantly larger than in the BCB-negative (BCB−) group. Immediately after oocyte selection according to BCB labelling, we found significant difference in chromatin configuration between the analyzed groups. BCB+ oocytes were significantly better at maturation than BCB− oocytes. BCB+ embryos were significantly more competent at cleaving and in their ability to reach the blastocyst stage than BCB− embryos. Ultrastructural analyses showed that the formation of active nucleoli in the BCB+ group started at the 8-cell stage. Conversely, most BCB− embryos at the 8-cell and 16-cell stages were fragmented. No statistically significant difference in RPC in nucleolus precursor bodies (NPBs) between BCB+ and BCB− oocytes was found. We can conclude that BCB labelling could be suitable for assessing the quality of porcine oocytes. Moreover, the evaluation of RPC indicates that the quantitative content of proteins in NPB is already established in growing oocytes.

Keywords

Brilliant cresyl blueIntranuclear characteristicsNucleoliNucleologenesis of embryosPig
Type
Research Article
Information
Zygote , Volume 27 , Issue 4 , August 2019 , pp. 232 - 240
Copyright
© Cambridge University Press 2019 

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