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dc.contributor.authorM. Vitale, Alejandra
dc.contributor.authorE. Kennedy Calvert, Meredith
dc.contributor.authorMallavarapu, Mallika
dc.contributor.authorYurttas, Piraye
dc.contributor.authorPerlin, Julie
dc.contributor.authorHerr, John
dc.contributor.authorCoonrod, Scott
dc.date.accessioned2017-05-03T16:57:36Z
dc.date.available2017-05-03T16:57:36Z
dc.date.issued2007
dc.date.modified2010-01-11T07:55:35Z
dc.identifier.issn1040452X
dc.identifier.doi10.1002/mrd.20648
dc.identifier.urihttp://hdl.handle.net/10072/28198
dc.description.abstractIn an effort to better understand oocyte function, we utilized two-dimensional (2D) electrophoresis and mass spectrometry to identify proteins that are differentially expressed during murine oocyte maturation. Proteins from 500 germinal vesicle (GV) and metaphase II-(MII) arrested oocytes were extracted, resolved on 2D electrophoretic gels, and stained with silver. Analysis of the gels indicated that 12 proteins appeared to be differentially expressed between the GV and MII stage. These proteins were then cored from the 2D gels and identified by mass spectrometry as: transforming acidic coiled-coil protein 3 (TACC3), heat shock protein 105 (HSP105), programmed cell death six-interacting protein (PDCD6IP), stress-inducible phosphoprotein (STI1), importin 2, adenylsuccinate synthase (ADDS), nudix, spindlin, lipocalin, lysozyme, translationally controlled tumor protein (TCTP), and nucleoplasmin 2 (NPM2). Interestingly, PDCD6IP, importin 2, spindlin, and NPM2 appear slightly larger in mass and more acidic on the MII oocyte gel compared to the GV oocyte gel, suggesting that they may be post-translationally modified during oocyte maturation. Given NPM2 is an oocyte-restricted protein, we chose to further investigate its properties during oocyte maturation and preimplantation development. Real-Time RT-PCR showed that NPM2 mRNA levels rapidly decline at fertilization. Indirect immunofluorescence analysis showed that, with the exception of cortical localization in MII-arrested oocytes, NPM2 is localized to the nucleus of both GV stage oocytes and all stages of preimplantation embryos. We then performed one-dimensional (1D) western blot analysis of mouse oocytes and preimplantation embryos and found that, as implicated by the 2D gel comparison, NPM2 undergoes a phosphatase-sensitive electrophoretic mobility shift during the GV to MII transition. The slower migrating NPM2 form is also present in pronuclear embryos but by the two-cell stage, the majority of NPM2 exists as the faster migrating form, which persists to the blastocyst stage.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherJohn Wiley & Sons
dc.publisher.placeUSA
dc.relation.ispartofstudentpublicationN
dc.relation.ispartofpagefrom608
dc.relation.ispartofpageto616
dc.relation.ispartofissue5
dc.relation.ispartofjournalMolecular Reproduction and Development
dc.relation.ispartofvolume74
dc.rights.retentionY
dc.subject.fieldofresearchEndocrinology
dc.subject.fieldofresearchBiochemistry and Cell Biology
dc.subject.fieldofresearchPaediatrics and Reproductive Medicine
dc.subject.fieldofresearchcode110306
dc.subject.fieldofresearchcode0601
dc.subject.fieldofresearchcode1114
dc.titleProteomic profiling of murine oocyte maturation
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.date.issued2007
gro.hasfulltextNo Full Text
gro.griffith.authorVitale, Alejandra M.


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