title: Identification of new ovulation-related genes in humans by comparing the transcriptome of granulosa cells before and after ovulation triggering in the same controlled ovarian stimulation cycle.
abstract: OBJECTIVE
  Which genes and molecular mechanisms are involved in the human ovulatory cascade and final oocyte maturation?
  
  
  CONCLUSIONS
  Up-regulated genes in granulosa cells (GC) represented inflammation, angiogenesis, extracellular matrix, growth factors and genes previously associated with ovarian cancer, while down-regulated genes mainly represented cell cycle and proliferation.
  
  
  BACKGROUND
  Radical changes occur in the follicle during final follicle maturation after the ovulatory trigger: these range from ensuring an optimal milieu for the oocyte in meiotic arrest to the release of a mature oocyte and remodeling into a corpus luteum. A wide range of mediators of final follicle maturation has been identified in rodents, non-human primates and cows.
  
  
  METHODS
  Prospective cohort study including 24 women undergoing ovarian stimulation with the long gonadotrophin-releasing hormone agonist protocol during 2010-2012 at Holbæk Fertility Clinic. Nine paired samples of GC and 24 paired samples of follicular fluid (FF) were obtained before and after recombinant human chorionic gonadotrophin (rhCG) administration.
  
  
  METHODS
  Nine paired (nine arrays before rhCG and nine arrays after rhCG) samples of GC mRNA were amplified and hybridized to Affymetrix Human Gene 1.0 ST GeneChip arrays, compared and bioinformatically analyzed. Eleven selected genes were validated by quantitative reverse transcriptase PCR. FF hormones were analyzed by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  Eleven hundred and eighty-six genes were differentially expressed (>2-fold, P<0.0001, false discovery rate <0.0012) when comparing GC isolated before and 36 h after hCG, among those were genes known to be expressed at ovulation, i.e. ADAMTS1 and HAS2. Many new ovulation-related genes were revealed, such as CD24, ANKRD22, CLDN11 and FBXO32. FF estrogen, androstenedione and anti-Müllerian hormone decreased significantly while progesterone increased, accompanied by radical changes in the expression of steroidogenic genes (CYP17A, CYP19A, HSD11B1 and HSD11B2, StAR). Genes related to inflammation, angiogenesis, extracellular matrix formation, growth factors and cancer were up-regulated while cell cycle genes were massively down-regulated. Seventy-two genes previously described in connection with ovarian cancer were among the highly regulated genes. In silico analysis for top upstream regulators of the ovulatory trigger suggested--besides LH--TNF, IGF1, PGR, AR, EGR1 (early growth response 1), ERK1/2 (extracellular signal regulated kinase 1/2) and CDKN1A (cyclin-dependent kinase inhibitor 1A) as potential mediators of the LH/hCG response.
  
  
  CONCLUSIONS
  The present dataset was generated from women under hormonal stimulation. However, comparison with a macaque natural cycle whole follicle ovulation dataset revealed major overlap, supporting the idea that the ovulation-related genes found in this study are relevant in the human natural cycle.
  
  
  CONCLUSIONS
  These data will serve as a research resource for genes involved in human ovulation and final oocyte maturation. Ovulation-related genes might be good candidate biomarkers of follicle and oocyte health. Further, some of the ovulation-related genes may serve as future ovarian cancer biomarkers.
  
  
  BACKGROUND
  Grants from the Research Fund of Region Sjælland are gratefully acknowledged. None of the authors declared any conflict of interest.
  
  
  BACKGROUND
  Not applicable.
authors: The Fertility Clinic, Holbæk Sygehus, Holbæk, Denmark.
authors: M L
authors: ML
authors: Wissing
authors: S G
authors: SG
authors: Kristensen
authors: C Y
authors: CY
authors: Andersen
authors: A L
authors: AL
authors: Mikkelsen
authors: T
authors: T
authors: Høst
authors: R
authors: R
authors: Borup
authors: M L
authors: ML
authors: Grøndahl
mesh_terms: D000328:Adult; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D010060:Ovulation / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D010062:Ovulation Induction; D011446:Prospective Studies; D059467:Transcriptome
chemical_list: D006063:Chorionic Gonadotropin
keywords: follicular fluid; granulosa cell transcriptome; hCG trigger; human; ovulation