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Expression and cellular distribution of zona pellucida glycoproteins in canine oocytes before and after in vitro maturation

Published online by Cambridge University Press:  15 October 2014

Bartosz Kempisty*
Affiliation:
Department of Histology and Embryology, Department of Anatomy, Poznan University of Medical Sciences, 6 Święcickiego St., 60–781 Poznań, Poland. Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland. Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
Hanna Piotrowska
Affiliation:
Department of Toxicology, Poznan University of Medical Sciences, Poznan, Poland.
Dorota Bukowska
Affiliation:
Institute of Veterinary Sciences, Poznan University of Life Sciences, Poznan, Poland.
Magdalena Woźna
Affiliation:
Institute of Veterinary Sciences, Poznan University of Life Sciences, Poznan, Poland.
Sylwia Ciesiółka
Affiliation:
Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
Katarzyna Wojtanowicz-Markiewicz
Affiliation:
Institute of Veterinary Sciences, Poznan University of Life Sciences, Poznan, Poland.
Renata Włodarczyk
Affiliation:
Institute of Veterinary Sciences, Poznan University of Life Sciences, Poznan, Poland.
Karol Jopek
Affiliation:
Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
Michal Jeseta
Affiliation:
Department of Genetics and Reproduction, Veterinary Research Institute, Brno, Czech Republic.
Małgorzata Bruska
Affiliation:
Department of Anatomy, Poznan University of Medical Sciences, Poznan, Poland.
Michał Nowicki
Affiliation:
Department of Histology and Embryology, Poznan University of Medical Sciences, Poznan, Poland.
Jędrzej M. Jaśkowski
Affiliation:
Institute of Veterinary Sciences, Poznan University of Life Sciences, Poznan, Poland.
Klaus-Peter Brüssow
Affiliation:
Institute of Reproductive Biology, Leibniz Institute for Farm Animal Biology, Wilhelm-Stahl-Allee 2, 18196 Dummerstorf, Germany.
Maciej Zabel
Affiliation:
Department of Histology and Embryology, Wroclaw Medical University, Wroclaw, Poland.
*
All correspondence to: Bartosz Kempisty. Department of Histology and Embryology, Department of Anatomy, Poznan University of Medical Sciences, 6 Święcickiego St., 60–781 Poznań, Poland. Tel: +48 61 8546419. Fax: +48 61 8546455. e-mail: etok@op.pl

Summary

This study was aimed at investigating zona pellucida glycoproteins (ZP) ZP2, ZP3 mRNA expression as well as ZP3, ZP4 (ZPB) protein distribution before and after in vitro maturation (IVM) in canine oocytes. The cumulus–oocyte complexes (COCs) were recovered from 27 anoestrous mongrel bitches and matured for 72 h in TCM199 medium. The canine COCs were analysed before and after IVM. Using real-time quantitative polymerase chain reaction (RQ-PCR), both groups of oocytes were analysed for detection of ZP2 and ZP3 mRNA profiles as well as using confocal microscopic analysis for observation of ZP3 and ZP4 protein distribution. In post-IVM canine oocytes an increase in transcript content of ZP2 and ZP3 genes as well as a decrease in ZP3 and ZP4 protein levels were observed when compared with pre-IVM oocytes. Moreover, the ZP4 protein before IVM was significantly distributed in the peripheral area of cytoplasm, whereas after IVM it was localized rather than in the entire cytoplasm. In contrast, the ZP3 protein was found both before and after IVM was distributed in the peripheral area of the cytoplasm. In conclusion, we suggest that the expression of ZP2 and ZP3 genes is associated with the maturation stage of canine oocytes, as higher mRNAs levels were found after IVM. However, a decreased expression of ZP3 and ZP4 proteins after IVM suggests maturation-dependent down-regulation of these protein translations, which may result in disturbed fertilization.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2014 

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