Immunohistochemical localization of zona pellucida proteins ZPA, ZPB and ZPC in human; cynomolgus monkey; and mouse ovaries

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dc.contributor.authorEderspaecher, U.
dc.contributor.authorBecker, A.
dc.contributor.authorBringmann, P.
dc.contributor.authorvan der Merwe, L.
dc.contributor.authorDonner, P.
dc.contributor.departmentExperimental Biology Programme, MRC, Tygerberg, South Africaen_US
dc.date.accessioned2020-03-19T10:17:24Z
dc.date.available2020-03-19T10:17:24Z
dc.date.epub2001
dc.date.issued2001
dc.description.abstractThe zona pellucida of mammalian oocytes plays an important role in binding and activation of sperm cells during the molecular events leading to fertilization. The genes coding for the three zona pellucida glycoproteins ZPA, ZPB, and ZPC of various species including mouse, dog, and human have been cloned and sequenced by several groups. However, it has remained a matter of debate as to whether the oocytes alone or in conjunction with the surrounding granulosa cells express and deposit these proteins to form the zona pellucida matrix. Addressing this unresolved issue, we assessed the expression and localization of all three zona pellucida proteins in ovaries of human, cynomolgus monkey and mice using immunohistochemical methods. In addition, oocyte-specific expression of ZPC from the primordial stage onward was confirmed by in situ hybridization. In sections of human ovaries, ZPA, ZPB, and ZPC proteins were immunohistochemically detected in the cytoplasms of primordial oocytes and during later stages of folliculogenesis in the zona pellucida matrices of oocytes. In sections fixed with formalin, a clear homogeneous ring was visible around the oocyte and no staining of granulosa cells was observed. In contrast, staining of ZP proteins was also observed between granulosa cells when Bouin's reagent had been used for tissue fixation. Thus, the original zona pellucida architecture was better preserved by formalin fixation. We further demonstrated that dissolution of the zona pellucida of isolated bovine oocytes occurred after they were exposed to Bouin's reagent. In summary, these results demonstrate that in mice, monkeys and humans, zona proteins are expressed and assembled exclusively by the oocyte and not by the granulosa cells. Previously observed results of ZP expression by an involvement of granulosa cells might therefore be the result of an improper fixation of the tissues leading to the disruption of the zona pellucida. Additionally this study highlights the importance of choosing the correct fixative for immunohistochemistry, not only for the usual reason of retaining antigenicity, but rather to retain the entire architectural structure.en_US
dc.description.sponsorshipThe excellent technical support of Marina Isernhagen, Gisela Hübner-Kosney, Hye-Youn Lee, Michael Niesar, Silke Finster, Alexandra Liese, Oliver Schilling and Simone Joschko is gratefully acknowledged.en_US
dc.identifier.citationEberspaecher U., Becker A., Bringmann P., van der Merwe L., Donner P. Immunohistochemical localization of zona pellucida proteins ZPA, ZPB and ZPC in human, cynomolgus monkey and mouse ovaries.en_US
dc.identifier.journalCell Tissue Resen_US
dc.identifier.urihttps://infospace.mrc.ac.za/handle/11288/595231
dc.language.isoenen_US
dc.publisherSpringeren_US
dc.relation.urlhttps://doi.org/10.1007/s004410000287en_US
dc.research.unitBiomedical Research and Innovation Platformen_US
dc.rightsAttribution 3.0 United States*
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/us/*
dc.subjectZona Pellucidaen_US
dc.subjectImmunohistochemistryen_US
dc.subjectIn situ hybridizationen_US
dc.subjectLocalizationen_US
dc.subjectFixationen_US
dc.subjectHumanen_US
dc.subjectHuman, macaca fascicularis, Mouse(NMRI)en_US
dc.titleImmunohistochemical localization of zona pellucida proteins ZPA, ZPB and ZPC in human; cynomolgus monkey; and mouse ovariesen_US
dc.typeArticleen_US
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