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Intranuclear characteristics of pig oocytes stained with brilliant cresyl blue and nucleologenesis of resulting embryos.
Zygote 2019; 27(4):232-240Z

Abstract

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.

Authors+Show Affiliations

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.Constantine the Philosopher University in Nitra, Faculty of Natural Sciences, Nabrezie mladeze 91, 94974 Nitra, Slovak Republic.Constantine the Philosopher University in Nitra, Faculty of Natural Sciences, Nabrezie mladeze 91, 94974 Nitra, Slovak Republic.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.Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Rumburska 89, 277 21 Libechov, Czech Republic.Friedrich Loeffler Institute, Institute of Farm Animal Genetics, Holtystrasse 10, D-31535 Neustadt, Germany.Ss. Cyril and Methodius University in Skopje, Republic of Macedonia.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.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31397243

Citation

Murin, Matej, et al. "Intranuclear Characteristics of Pig Oocytes Stained With Brilliant Cresyl Blue and Nucleologenesis of Resulting Embryos." Zygote (Cambridge, England), vol. 27, no. 4, 2019, pp. 232-240.
Murin M, Strejcek F, Bartkova A, et al. Intranuclear characteristics of pig oocytes stained with brilliant cresyl blue and nucleologenesis of resulting embryos. Zygote. 2019;27(4):232-240.
Murin, M., Strejcek, F., Bartkova, A., Morovic, M., Benc, M., Prochazka, R., ... Laurincik, J. (2019). Intranuclear characteristics of pig oocytes stained with brilliant cresyl blue and nucleologenesis of resulting embryos. Zygote (Cambridge, England), 27(4), pp. 232-240. doi:10.1017/S0967199419000352.
Murin M, et al. Intranuclear Characteristics of Pig Oocytes Stained With Brilliant Cresyl Blue and Nucleologenesis of Resulting Embryos. Zygote. 2019;27(4):232-240. PubMed PMID: 31397243.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Intranuclear characteristics of pig oocytes stained with brilliant cresyl blue and nucleologenesis of resulting embryos. AU - Murin,Matej, AU - Strejcek,Frantisek, AU - Bartkova,Alexandra, AU - Morovic,Martin, AU - Benc,Michal, AU - Prochazka,Radek, AU - Lucas-Hahn,Andrea, AU - Pendovski,Lazo, AU - Laurincik,Jozef, Y1 - 2019/08/09/ PY - 2019/8/10/pubmed PY - 2019/8/10/medline PY - 2019/8/10/entrez KW - Brilliant cresyl blue KW - Intranuclear characteristics KW - Nucleoli KW - Nucleologenesis of embryos KW - Pig SP - 232 EP - 240 JF - Zygote (Cambridge, England) JO - Zygote VL - 27 IS - 4 N2 - 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. SN - 1469-8730 UR - https://www.unboundmedicine.com/medline/citation/31397243/Intranuclear_characteristics_of_pig_oocytes_stained_with_brilliant_cresyl_blue_and_nucleologenesis_of_resulting_embryos L2 - https://www.cambridge.org/core/product/identifier/S0967199419000352/type/journal_article DB - PRIME DP - Unbound Medicine ER -