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Calreticulin from suboolemmal vesicles affects membrane regulation of polyspermy

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dc.contributor.advisorRomar Andrés, Raquel
dc.contributor.authorSaavedra MD
dc.contributor.authorCoy, Pilar
dc.contributor.authorBetancourt, M
dc.contributor.authorGonzález-Marquez, H
dc.contributor.authorMondéjar Corbalán, Irene
dc.contributor.authorJíménez Movilla, María
dc.contributor.authorAvilés Sánchez, Manuel
dc.contributor.departmentFisiología
dc.contributor.otherFacultades de la UMU
dc.date.accessioned2026-03-30T10:32:41Z
dc.date.available2026-03-30T10:32:41Z
dc.date.copyright©2014 Society for Reproduction and Fertility
dc.date.issued2014-02-05
dc.description.abstractThis study was designed to determine whether calreticulin (CRT), a chaperone protein, is present in in vitro-matured (IVM) pig oocytes and to study its potential role in the block to polyspermy. Western blot analysis, using an anti-CRT antibody, of oocyte lysate showed an immunoreactive band of w60 kDa. Simultaneous labeling of IVM oocytes with anti-CRT antibody and peanut agglutinin lectin (PNA lectin, a porcine cortical granules (CG)-specific binding lectin) revealed localization of CRT in the subplasmalemmal region with a 27.7% colocalization with PNA staining. After IVF, PNA labeling was not observed and anti-CRT labeling decreased significantly in zygotes and disappeared in two-cell embryos. Western blot analysis of oocyte exudate obtained from zona pellucida (ZP)-free oocytes activated with calcium ionophore confirmed the presence of a band that reacted with an anti-CRT antibody. Anti-CRT antibody and PNA labeling were not observed in activated oocytes despite being detectable in non-activated oocytes. The presence of CRT in vesicles located under the oolemma was demonstrated using immunogold cytochemistry at the ultrastructural level. To study the role of CRT in fertilization, ZP-enclosed and ZP-free oocytes were incubated with exogenous CRT and then inseminated. Whereas ZP-free oocytes showed fewer penetrating sperm and lower polyspermy rates than untreated oocytes, the opposite effect was observed in ZP-enclosed oocytes. In conclusion, CRT is confined to subplasmalemmal vesicles partially overlapping with CG contents. Its exocytosis after the oocyte activation seems to participate in the membrane block to polyspermy in pigs but is not involved in the ZP block.
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dc.identifier.citationSaavedra MD, Mondéjar I, Coy P, Betancourt M, González-Márquez H, Jiménez-Movilla M, Avilés M, Romar R. Calreticulin from suboolemmal vesicles affects membrane regulation of polyspermy. Reproduction. 2014 Feb 5;147(3):369-78. doi: 10.1530/REP-13-0454. PMID: 24398873.
dc.identifier.doi10.1530/REP-13-0454
dc.identifier.eissn1741-7899
dc.identifier.urihttp://hdl.handle.net/10201/224181
dc.languageeng
dc.relationThis work was supported by MEC and FEDER (grant AGL2009-12512-C02-01) and MINECO (grant AGL2012-40180-C03-01-02).
dc.relation.publisherversionhttps://academic.oup.com/reproduction/article/147/3/369/8235807
dc.rights.accessRightsinfo:eu-repo/semantics/restrictedAccess
dc.subject.odsNo relacionado con ningún objetivo de desarrollo sostenible
dc.titleCalreticulin from suboolemmal vesicles affects membrane regulation of polyspermy
dc.typeinfo:eu-repo/semantics/article
dc.type.versioninfo:eu-repo/semantics/publishedVersion
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