Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/83268
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dc.contributor.author | Frank, L. | - |
dc.contributor.author | Sutton-McDowall, M. | - |
dc.contributor.author | Brown, H. | - |
dc.contributor.author | Russell, D. | - |
dc.contributor.author | Gilchrist, R. | - |
dc.contributor.author | Thompson, J. | - |
dc.date.issued | 2014 | - |
dc.identifier.citation | Human Reproduction, 2014; 29(6):1292-1303 | - |
dc.identifier.issn | 0268-1161 | - |
dc.identifier.issn | 1460-2350 | - |
dc.identifier.uri | http://hdl.handle.net/2440/83268 | - |
dc.description.abstract | STUDY QUESTION What is the effect of beta-O-linked glycosylation (O-GlcNAcylation) on specific proteins in the cumulus-oocyte complex (COC) under hyperglycaemic conditions? SUMMARY ANSWER Heat shock protein 90 (HSP90) was identified and confirmed as being O-GlcNAcylated in mouse COCs under hyperglycaemic conditions (modelled using glucosamine), causing detrimental outcomes for embryo development. WHAT IS KNOWN ALREADY O-GlcNAcylation of proteins occurs as a result of increased activity of the hexosamine biosynthesis pathway, which provides substrates for cumulus matrix production during COC maturation, and also for O-GlcNAcylation. COCs matured under hyperglycaemic conditions have decreased developmental competence, mediated at least in part through the mechanism of increased O-GlcNAcylation. STUDY DESIGN, SIZE, DURATION This study was designed to examine the effect of hyperglycaemic conditions (using the hyperglycaemic mimetic, glucosamine) on O-GlcNAc levels in the mouse COC, and furthermore to identify potential candidate proteins which are targets of this modification, and their roles in oocyte maturation. PARTICIPANTS/MATERIALS, SETTING, METHODS COCs from 21-day-old superovulated CBA × C57BL6 F1 hybrid female mice were matured in vitro (IVM). Levels of O-GlcNAcylated proteins, HSP90 and O-GlcNAc transferase (OGT, the enzyme responsible for O-GlcNAcylation) in COCs were measured using western blot, and localization observed using immunocytochemistry. For glycosylated HSP90 levels, and to test OGT-HSP90 interaction, immunoprecipitation was performed prior to western blotting. Embryo development was assessed using in vitro fertilization and embryo culture post-maturation. MAIN RESULTS AND THE ROLE OF CHANCE Addition of the hyperglycaemic mimetic glucosamine to IVM medium for mouse COCs increased detectable O-GlcNAcylated protein levels (by western blot and immunocytochemistry), and this effect was reversed using an OGT inhibitor (P < 0.05). HSP90 was identified as a target of O-GlcNAcylation in the COC, and inhibition of HSP90 during IVM reversed glucosamine-induced decreases in oocyte developmental competence (P < 0.05). We also demonstrated the novel finding of an association between HSP90 and OGT in COCs, suggesting a possible client–chaperone relationship. LIMITATIONS, REASONS FOR CAUTION In vitro maturation of COCs was used so that treatment time could be limited to the 17 h of maturation prior to ovulation. Additionally, glucosamine, a hyperglycaemic mimetic, was used because it specifically activates the hexosamine pathway which provides the O-GlcNAc moieties. The results in this study should be confirmed using in vivo models of hyperglycaemia and different HSP90 inhibitors. WIDER IMPLICATIONS OF THE FINDINGS This study leads to a new understanding of how diabetes influences oocyte competence and provides insight into possible therapeutic interventions based on inhibiting HSP90 to improve oocyte quality. | - |
dc.description.statementofresponsibility | L.A. Frank, M.L. Sutton-McDowall, H.M. Brown, D.L. Russell, R.B. Gilchrist, and J.G. Thompson | - |
dc.language.iso | en | - |
dc.publisher | Oxford University Press | - |
dc.rights | © The Author 2014. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com | - |
dc.source.uri | http://dx.doi.org/10.1093/humrep/deu066 | - |
dc.subject | Hyperglycaemia | - |
dc.subject | oocyte developmental competence | - |
dc.subject | hexosamine pathway | - |
dc.subject | HSP90 | - |
dc.subject | O-GlcNAc | - |
dc.title | Hyperglycaemic conditions perturb mouse oocyte in vitro developmental competence via beta-O-linked glycosylation of Heat shock protein 90 | - |
dc.type | Journal article | - |
dc.contributor.organisation | Robinson Institute | - |
dc.identifier.doi | 10.1093/humrep/deu066 | - |
dc.relation.grant | http://purl.org/au-research/grants/nhmrc/453556 | - |
pubs.publication-status | Published | - |
dc.identifier.orcid | Sutton-McDowall, M. [0000-0002-4121-0202] | - |
dc.identifier.orcid | Brown, H. [0000-0001-6342-3316] | - |
dc.identifier.orcid | Russell, D. [0000-0002-4930-7658] | - |
dc.identifier.orcid | Gilchrist, R. [0000-0003-1611-7142] | - |
dc.identifier.orcid | Thompson, J. [0000-0003-4941-7731] | - |
Appears in Collections: | Aurora harvest 4 Obstetrics and Gynaecology publications |
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hdl_83268.pdf | Accepted version | 1.58 MB | Adobe PDF | View/Open |
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