title: TGFβ (transforming growth factor β) superfamily members and their receptors in the fetal porcine ovaries: effect of prenatal flutamide treatment.
abstract: BACKGROUND
  We have recently demonstrated that antiandrogen treatment during fetal life resulted in delayed folliculogenesis. The aim of the present study was to investigate the effect of androgen deficiency induced by flutamide on the expression of TGFβ superfamily members and their receptors which are involved in follicle formation and its transition to the primary stage.
  
  
  METHODS
  Pregnant gilts were injected with flutamide (for seven days, 50 mg/day/kg b.w.) or corn oil (controls) starting at 43 (GD50), 83 (GD90) or 101 (GD108) gestational day. The expression in fetal ovaries of selected TGFβ superfamily members (AMH, BMP4, GDF9), their receptors (AMHR-II, BMPR-IB, BMPR-II), and Smad1 and Smad3 proteins involved in signal transduction were investigated by real-time PCR and/or immunohistochemistry.
  
  
  RESULTS
  Flutamide treatment increased the expression of BMP4 mRNA on GD50 and GD108 and BMPR-IB mRNA on GD50. The expression of BMPR-II was decreased at mRNA level and lower immunostaining intensity was observed after flutamide administration only on GD50. GDF9 and AMHR-II mRNA expression levels were significantly downregulated in both GD90 and GD108 groups. However, AMHR-II was immunolocalized only on GD108 and less positively stained oocytes were found after flutamide treatment. AMH mRNA level was diminished in the GD90 group, while it was elevated in the GD108 group. Moreover, the higher amounts of positively stained oocytes for phosphorylated form of Smad1 were observed following flutamide administration on GD108.
  
  
  CONCLUSIONS
  Experimentally-induced androgen deficiency during fetal development deregulates the expression level of some of TGFβ superfamily members and their receptors which may affect primordial follicle assembly. Our findings further underline the role of androgens in the early stages of follicle development.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Knapczyk-Stwora
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Grzesiak
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Duda
authors.forename: Marek
authors.initials: M
authors.lastname: Koziorowski
authors.forename: Jerzy
authors.initials: J
authors.lastname: Galas
authors.affiliation: Department of Endocrinology, Institute of Zoology, Jagiellonian University in Krakow, Poland. maria.slomczynska@uj.edu.pl.
authors.forename: Maria
authors.initials: M
authors.lastname: Slomczynska
mesh_terms: D000726:Androgen Antagonists / Q000494:pharmacology; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics / Q000378:metabolism; D005260:Female; D005485:Flutamide / Q000494:pharmacology*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D010053:Ovary / Q000187:drug effects* / Q000648:ultrastructure; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism*; D013552:Swine; D016212:Transforming Growth Factor beta / Q000235:genetics*
chemical_list: D000726:Androgen Antagonists; D055415:Bone Morphogenetic Protein 4; D012333:RNA, Messenger; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; D005485:Flutamide