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Type: Journal article
Title: Microarray analysis of mRNA from cumulus cells following in vivo or in vitro maturation of mouse cumulus-oocyte complexes
Author: Kind, K.
Banwell, K.
Gebhardt, K.
Macpherson, A.
Gauld, A.
Russell, D.
Thompson, J.
Citation: Reproduction Fertility and Development, 2013; 25(2):426-438
Publisher: C S I R O Publishing
Issue Date: 2013
ISSN: 1031-3613
Statement of
Karen L. Kind, Kelly M. Banwell, Kathryn M. Gebhardt, Anne Macpherson, Ashley Gauld, Darryl L. Russell and Jeremy G. Thompson
Abstract: The IVM of mammalian cumulus–oocyte complexes (COCs) yields reduced oocyte developmental competence compared with oocytes matured in vivo. Altered cumulus cell function during IVM is implicated as one cause for this difference. We have conducted a microarray analysis of cumulus cell mRNA following IVM or in vivo maturation (IVV). Mouse COCs were sourced from ovaries of 21-day-old CBAB6F1 mice 46 h after equine chorionic gonadotrophin (5 IU, i.p.) or from oviducts following treatment with 5 IU eCG (61 h) and 5 IU human chorionic gonadotrophin (13 h). IVM was performed in α-Minimal Essential Medium with 50 mIU FSH for 17 h. Three independent RNA samples were assessed using the Affymetrix Gene Chip Mouse Genome 430 2.0 array (Affymetrix, Santa Clara, CA, USA). In total, 1593 genes were differentially expressed, with 811 genes upregulated and 782 genes downregulated in IVM compared with IVV cumulus cells; selected genes were validated by real-time reverse transcription–polymerase chain reaction (RT-PCR). Surprisingly, haemoglobin α (Hba-a1) was highly expressed in IVV relative to IVM cumulus cells, which was verified by both RT-PCR and western blot analysis. Because haemoglobin regulates O2 and/or nitric oxide availability, we postulate that it may contribute to regulation of these gases during the ovulatory period in vivo. These data will provide a useful resource to determine differences in cumulus cell function that are possibly linked to oocyte competence.
Keywords: Affymetrix; gene expression; haemoglobin
Rights: Journal compilation © CSIRO 2012
RMID: 0020122435
DOI: 10.1071/RD11305
Appears in Collections:Obstetrics and Gynaecology publications
IPAS publications

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