Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/8034
Citations
Scopus Web of Science® Altmetric
?
?
Full metadata record
DC FieldValueLanguage
dc.contributor.authorGilchrist, R.en
dc.contributor.authorRitter, L.en
dc.contributor.authorCranfield, M.en
dc.contributor.authorJeffrey, L.en
dc.contributor.authorAmato, F.en
dc.contributor.authorScott, S.en
dc.contributor.authorMyllymaa, S.en
dc.contributor.authorKaivo-Oja, N.en
dc.contributor.authorLankinen, H.en
dc.contributor.authorMottershead, D.en
dc.contributor.authorGroome, N.en
dc.contributor.authorRitvos, O.en
dc.date.issued2004en
dc.identifier.citationBiology of Reproduction, 2004; 71(3):732-739en
dc.identifier.issn0006-3363en
dc.identifier.issn1529-7268en
dc.identifier.urihttp://hdl.handle.net/2440/8034-
dc.description© 2004 by the Society for the Study of Reproduction, Inc.en
dc.description.abstractParacrine factors secreted by oocytes play a pivotal role in promoting early ovarian follicle growth and in defining a morphogenic gradient in antral follicles, yet the exact identities of these oocyte factors remain unknown. This study was conducted to determine the extent to which the mitogenic activity of mouse oocytes can be attributed to growth differentiation factor 9 (GDF9). To do this, specific anti-human GDF9 monoclonal antibodies were generated. Based on epitope mapping and bioassays, a GDF9 neutralizing antibody, mAb-GDF9-53, was characterized with very low cross-reactivity with related transforming growth factor (TGF)ß superfamily members, including BMP15 (also called GDF9B). Pep-SPOT epitope mapping showed that mAb-GDF9-53 recognizes a short 4-aa sequence, and three-dimensional peptide modeling suggested that this binding motif lies at the C-terminal fingertip of mGDF9. As predicted by sequence alignments and modeling, the antibody detected recombinant GDF9, but not BMP15 in a Western blot and GDF9 protein in oocyte extract and oocyte-conditioned medium. In a mouse mural granulosa cell (MGC) bioassay, mAb-GDF9-53 completely abolished the mitogenic effects of GDF9, but had no effect on TGFß1 or activin A-stimulated MGC proliferation. An unrelated IgG at the same dose had no effect on GDF9 activity. This GDF9 neutralizing antibody was then tested in an established oocyte-secreted mitogen bioassay, where denuded oocytes cocultured with granulosa cells promote cell proliferation in a dose-dependent manner. The mAb-GDF9-53 dose dependently (0–160 µg/ml) decreased the mitogenic activity of oocytes but only by 45% at the maximum dose of mAb. Just 5 µg/ml of mAb-GDF9-53 neutralized 90% of recombinant mGDF9 mitogenic activity, but only 15% of oocyte activity. Unlike mAb-GDF9-53, a TGFß pan-specific neutralizing antibody did not affect the mitogenic capacity of the oocyte, but completely neutralized TGFß1-induced DNA synthesis. This study has characterized a specific GDF9 neutralizing antibody. Our data provide the first direct evidence that the endogenous GDF9 protein is an important oocyte-secreted mitogen, but also show that GDF9 accounts for only part of total oocyte bioactivity.en
dc.description.statementofresponsibilityR.B. Gilchrist, L.J. Ritter, M. Cranfield, L.A. Jeffery, F. Amato, S.J. Scott, S. Myllymaa, N. Kaivo-Oja, H. Lankinen, D.G. Mottershead, N.P. Groome, and O. Ritvosen
dc.language.isoenen
dc.publisherSoc Study Reproductionen
dc.subjectOocytes; Animals; Mice; Intercellular Signaling Peptides and Proteins; Transforming Growth Factor beta; Mitogens; Antibodies, Monoclonal; Amino Acid Sequence; Protein Structure, Tertiary; Molecular Sequence Data; Female; Growth Differentiation Factor 9; Bone Morphogenetic Protein 15en
dc.titleImmunoneutralization of growth differentiation factor 9 reveals it partially accounts for mouse oocyte mitogenic activityen
dc.typeJournal articleen
dc.identifier.rmid0020041225en
dc.identifier.doi10.1095/biolreprod.104.028852en
dc.identifier.pubid56463-
pubs.library.collectionObstetrics and Gynaecology publicationsen
pubs.verification-statusVerifieden
pubs.publication-statusPublisheden
dc.identifier.orcidGilchrist, R. [0000-0003-1611-7142]en
dc.identifier.orcidRitter, L. [0000-0001-5942-851X]en
Appears in Collections:Obstetrics and Gynaecology publications

Files in This Item:
There are no files associated with this item.


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.