title: Direct actions of androgens on the survival, growth and secretion of steroids and anti-Müllerian hormone by individual macaque follicles during three-dimensional culture.
abstract: OBJECTIVE
  What are the direct effects of androgens on primate follicular development and function at specific stages of folliculogenesis?
  
  
  CONCLUSIONS
  Androgen addition altered primate follicle survival, growth, steroid and anti-Müllerian hormone (AMH) production, and oocyte quality in vitro, in a dose- and stage-dependent manner.
  
  
  BACKGROUND
  Androgens have local actions in the ovary, particularly in the developing follicles. It is hypothesized that androgen promotes early follicular growth, but becomes detrimental to the antral follicles in primates.
  
  
  METHODS
  In vitro follicle maturation was performed using rhesus macaques. Secondary (125-225 µm) follicles were mechanically isolated from 14 pairs of ovaries, encapsulated into alginate (0.25% w/v), and cultured for 40 days.
  
  
  METHODS
  Individual follicles were cultured in a 5% O2 environment, in alpha minimum essential medium supplemented with recombinant human FSH. Follicles were randomly assigned to experiments of steroid ablation by trilostane (TRL), testosterone (T) replacement and dihydrotestosterone (DHT) replacement. Follicle survival and growth were assessed. Follicles with diameters ≥500 μm at Week 5 were categorized as fast-grow follicles. Pregnenolone (P5), progesterone (P4), estradiol (E2) and AMH concentrations in media were measured. Meiotic maturation and fertilization of oocytes from recombinant human chorionic gonadotrophin-treated follicles were assessed at the end of culture.
  
  
  RESULTS
  Compared with controls, TRL exposure reduced (P < 0.05) follicle survival, antrum formation rate and follicle diameters at Week 5. While P5 concentrations increased (P < 0.05) following TRL treatment, P4 levels decreased (P < 0.05) in fast-grow follicles at Week 5. Few healthy oocytes were retrieved from antral follicles developed in the presence of TRL. T replacement with TRL increased (P < 0.05) follicle survival and antrum formation at Week 5, compared with TRL alone, to levels comparable to controls. However, high-dose T with TRL decreased (P < 0.05) diameters of fast-grow follicles. Although P4 concentrations produced by fast-grow follicles were not altered by T in the presence of TRL, there was a dose-dependent increase (P < 0.05) in E2 levels at Week 5. High-dose T with TRL decreased (P < 0.05) AMH production by fast-grow follicles at Week 3. More healthy oocytes were retrieved from antral follicles developed in TRL+T compared with TRL alone. DHT had the similar effects to those of high-dose T, except that DHT replacement decreased (P < 0.05) E2 concentrations produced by fast-grow follicles at Week 5 regardless of TRL treatment.
  
  
  CONCLUSIONS
  This study reports T and DHT actions on in vitro-developed individual primate (macaque) follicles, which are limited to the interval from the secondary to small antral stage.
  
  
  CONCLUSIONS
  The above findings provide novel information on the role(s) of androgens in primate follicular development and oocyte maturation. We hypothesize that androgens promote pre-antral follicle development, but inhibit antral follicle growth and function in primates. While androgens can act positively, excess levels of androgens may have negative impacts on primate folliculogenesis.
  
  
  BACKGROUND
  NIH U54 RR024347/RL1HD058294/PL1EB008542 (Oncofertility Consortium), NIH U54 HD071836 (SCCPIR), NIH ORWH/NICHD 2K12HD043488 (BIRCWH), NIH FIC TW/HD-00668, ONPRC 8P51OD011092. There are no conflicts of interest.
authors.affiliation: Division of Reproductive & Developmental Sciences, Oregon National Primate Research Center, 505 NW 185th Avenue, Beaverton, OR 97006, USA Department of Gynecology and Obstetrics, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Av. Bandeirantes 3900, Monte Alegre, CEP, 14049, Ribeirão Preto, São Paulo, Brazil Departamento de Pesquisa e Desenvolvimento, Pró-Criar Medicina Reprodutiva, Rua Bernardo Guimarães 2063, Lourdes, 30140, Belo Horizonte, Minas Gerais, Brazil.
authors.forename: J K
authors.initials: JK
authors.lastname: Rodrigues
authors.affiliation: Department of Gynecology and Obstetrics, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Av. Bandeirantes 3900, Monte Alegre, CEP, 14049, Ribeirão Preto, São Paulo, Brazil.
authors.forename: P A
authors.initials: PA
authors.lastname: Navarro
authors.affiliation: Division of Reproductive & Developmental Sciences, Oregon National Primate Research Center, 505 NW 185th Avenue, Beaverton, OR 97006, USA Department of Obstetrics & Gynecology, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA.
authors.forename: M B
authors.initials: MB
authors.lastname: Zelinski
authors.affiliation: Division of Reproductive & Developmental Sciences, Oregon National Primate Research Center, 505 NW 185th Avenue, Beaverton, OR 97006, USA Department of Obstetrics & Gynecology, Oregon Health & Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA.
authors.forename: R L
authors.initials: RL
authors.lastname: Stouffer
authors.affiliation: Division of Reproductive & Developmental Sciences, Oregon National Primate Research Center, 505 NW 185th Avenue, Beaverton, OR 97006, USA xujin@ohsu.edu.
authors.forename: J
authors.initials: J
authors.lastname: Xu
mesh_terms: D000728:Androgens / Q000494:pharmacology*; D000818:Animals; D054304:Anti-Mullerian Hormone / Q000378:metabolism*; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D013196:Dihydrotestosterone / Q000031:analogs & derivatives / Q000494:pharmacology; D005260:Female; D059471:In Vitro Oocyte Maturation Techniques; D008251:Macaca; D006080:Ovarian Follicle / Q000187:drug effects* / Q000254:growth & development / Q000378:metabolism; D013739:Testosterone / Q000494:pharmacology
chemical_list: D000728:Androgens; D013196:Dihydrotestosterone; D013739:Testosterone; D054304:Anti-Mullerian Hormone; C009954:trilostane
keywords: androgens; dihydrotestosterone; follicle culture; follicle development; testosterone