title: Glycolytic enzyme expression in human granulosa cells.
abstract: OBJECTIVE
  To examine the differences in specific protein expression between mural and cumulus granulosa cells following 24-hour in vitro culture.
  
  
  METHODS
  Laboratory study.
  
  
  METHODS
  University Hospital.
  
  
  METHODS
  Human granulosa cells were collected at the time of egg collection during assisted reproduction. Cumulus cells associated with the oocyte were separated from mural cells from the periphery of the follicle before in vitro culture for 24 hours. Cells were then lysed and subjected to two-dimensional gel electrophoresis.
  
  
  METHODS
  Given that cumulus (cGC) and mural granulosa cells (mGC) differentiate from a single layer, it is likely that phenotypic differences between them may reflect specific molecular processes and structural adaptations. Computer-assisted analysis using dedicated software enabled the presence, absence, or relative volume of each individual protein spot to be estimated. Differentially expressed spots were identified using tandem mass spectrometry.
  
  
  RESULTS
  The mean number of separate protein spots detected in mGC gels was 1,105 +/- 146, and in cGC it was 887 +/- 236, although there was no statistically significant difference between the two. Five enzymes of the glycolytic pathway were never expressed in cGC after 24 hours in vitro; these were triose-phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase 1, and two isoforms of alpha enolase. These are the first data collected in humans consistent with a recent demonstration that isolated murine cGC cultured in vitro exhibit decreased expression of mRNA encoding glycolytic enzymes, and support the suggestion that some factor or factors secreted by the oocyte may be responsible for the maintenance of glycolysis in the adjacent cGC.
authors.affiliation: The London Bridge Fertility, Gynaecology & Genetics Centre, London Bridge, United Kingdom. dgillott@thebridgecentre.co.uk
authors.forename: David John
authors.initials: DJ
authors.lastname: Gillott
authors.forename: Abdallah
authors.initials: A
authors.lastname: Eldib
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Iammarrone
authors.forename: Kit-Yi
authors.initials: KY
authors.lastname: Leung
authors.forename: Alan Russell
authors.initials: AR
authors.lastname: Thornhill
authors.forename: Jurgis Gediminas
authors.initials: JG
authors.lastname: Grudzinskas
mesh_terms: D000293:Adolescent; D000328:Adult; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D015870:Gene Expression; D006019:Glycolysis; D006107:Granulosa Cells / Q000737:chemistry / Q000201:enzymology*; D006801:Humans