title: Pigment-Epithelium Derived Factor (PEDF) and the human ovary: a role in the generation of ROS in granulosa cells.
abstract: OBJECTIVE
  Pigment Epithelium Derived Factor (PEDF) is a multifunctional factor, which was found in mouse ovary and in human ovarian follicular fluid (FF). Its ovarian functions include anti-angiogenic actions. This study aimed to explore other PEDF-actions and the sites of PEDF expression in the human ovary.
  
  
  METHODS
  We used paraffin-embedded human ovarian sections for PEDF-immunohistochemistry and IVF-derived human granulosa cells (GCs) for RT-PCR, Western blotting and functional studies, including measurement of cell viability (ATP-assay), apoptosis (caspase-assay) and reactive oxygen species (ROS).
  
  
  RESULTS
  Immunohistochemistry revealed PEDF in the cytoplasm of GCs of avascular follicles from the preantral to the antral stage and in FF. PEDF was also found in luteinized GCs of the highly vascularized corpus luteum, a result not in line with a sole anti-angiogenic action. Like GCs in vivo, cultured human luteinizing GCs express PEDF. They also responded to exogenous recombinant PEDF. In low concentrations PEDF did not affect cell viability but caused generation ROS. ROS-induction by PEDF was a concentration-dependent process and may be due to the activity of NADPH oxidase (NOX) type 4 and/or 5, which as we found are expressed by GCs. An antioxidant and apocynin, which inhibits NOX, blocked ROS generation. High levels of exogenous recombinant PEDF induced apoptosis of GCs, which was prevented by antioxidants, implying involvement of ROS.
  
  
  CONCLUSIONS
  PEDF is emerging as an ovarian factor, which has unexpected ROS-augmenting activities in the human ovary. It may be involved in ovarian ROS homeostasis and may contribute to oxidative stress.
authors.affiliation: Anatomy III-Cell Biology, Ludwig-Maximilian-University (LMU), Munich, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Kampfer
authors.affiliation: Anatomy III-Cell Biology, Ludwig-Maximilian-University (LMU), Munich, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Saller
authors.affiliation: Anatomy III-Cell Biology, Ludwig-Maximilian-University (LMU), Munich, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Windschüttl
authors.affiliation: ART Bogenhausen, Munich, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Berg
authors.affiliation: ART Bogenhausen, Munich, Germany.
authors.forename: U
authors.initials: U
authors.lastname: Berg
authors.affiliation: Anatomy III-Cell Biology, Ludwig-Maximilian-University (LMU), Munich, Germany. Electronic address: Mayerhofer@lrz.uni-muenchen.de.
authors.forename: A
authors.initials: A
authors.lastname: Mayerhofer
mesh_terms: D000328:Adult; D017209:Apoptosis; D015153:Blotting, Western; D002470:Cell Survival; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005136:Eye Proteins / Q000008:administration & dosage / Q000378:metabolism*; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000378:metabolism; D000074663:NADPH Oxidase 4; D000074664:NADPH Oxidase 5; D019255:NADPH Oxidases / Q000378:metabolism; D009414:Nerve Growth Factors / Q000008:administration & dosage / Q000378:metabolism*; D010053:Ovary / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015843:Serpins / Q000008:administration & dosage / Q000378:metabolism*; D055815:Young Adult; D000099174:Pigment Epithelium-Derived Factor
chemical_list: D005136:Eye Proteins; D008565:Membrane Proteins; D000074663:NADPH Oxidase 4; D000074664:NADPH Oxidase 5; D019255:NADPH Oxidases; D009414:Nerve Growth Factors; D017382:Reactive Oxygen Species; D015843:Serpins; D000099174:Pigment Epithelium-Derived Factor; C498910:NOX4 protein, human; C431571:NOX5 protein, human
keywords: Growth factor; NADPH oxidase; Reproduction; Signaling