title: Microarray analysis of mRNA from cumulus cells following in vivo or in vitro maturation of mouse cumulus-oocyte complexes.
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 46h after equine chorionic gonadotrophin (5IU, i.p.) or from oviducts following treatment with 5IU eCG (61h) and 5IU human chorionic gonadotrophin (13h). IVM was performed in α-Minimal Essential Medium with 50 mIU FSH for 17h. 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.
authors.affiliation: Research Centre for Reproductive Health, The Robinson Institute, School of Paediatrics and Reproductive Health, The University of Adelaide, Adelaide, SA 5005, Australia.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Kind
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Banwell
authors.forename: Kathryn M
authors.initials: KM
authors.lastname: Gebhardt
authors.forename: Anne
authors.initials: A
authors.lastname: Macpherson
authors.forename: Ashley
authors.initials: A
authors.lastname: Gauld
authors.forename: Darryl L
authors.initials: DL
authors.lastname: Russell
authors.forename: Jeremy G
authors.initials: JG
authors.lastname: Thompson
mesh_terms: D000818:Animals; D015153:Blotting, Western; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D054885:Cumulus Cells / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D020869:Gene Expression Profiling; D006441:Hemoglobin A / Q000378:metabolism; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods; D051379:Mice; D046228:Microarray Analysis / Q000379:methods*; D009569:Nitric Oxide / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D009865:Oocytes / Q000378:metabolism / Q000502:physiology*; D010100:Oxygen / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D006063:Chorionic Gonadotropin; D012333:RNA, Messenger; D009569:Nitric Oxide; D006441:Hemoglobin A; D010100:Oxygen