Direct actions of androgens on the survival, growth and secretion of steroids and anti-Mü llerian hormone by individual macaque follicles during three-dimensional culture

studyquestion:What are thedirect effectsof androgensonprimatefollicular development andfunctionat specific stages of folliculogenesis? summaryanswer: 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. what is known already: 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. study design, size, duration: 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. participants/materials, setting, 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 μmatWeek 5 were categorized as fast-grow follicles. Pregnenolone (P5), progesterone (P4), estradiol (E2) and AMHconcentrations in media were measured. Meiotic maturation and fertilization of oocytes from recombinant human chorionic gonadotrophin-treated follicles were assessed at the end of culture. main results and the role of chance: Compared with controls, TRL exposure reduced (P <0.05) follicle survival, antrum formation rate and follicle diameters atWeek 5.While P5 concentrations increased (P <0.05) followingTRL treatment,P4 levelsdecreased (P 0.05) in fast-growfollicles atWeek 5. Fewhealthy oocyteswere retrieved fromantral follicles developed in the presence of TRL. T replacementwith TRL increased (P <0.05) follicle survival and antrum formation atWeek 5, comparedwith TRL alone, to levels comparable to controls.However, highdose Twith TRL decreased (P 0.05) diameters of fast-growfollicles. Although P4 concentrations produced by fast-growfollicleswere not altered by T in the presence of TRL, there was a dose-dependent increase (P <0.05) in E2 levels atWeek 5. High-dose T with TRL decreased (P 0.05) AMHproductionby fast-growfollicles atWeek 3.More healthyoocyteswere retrievedfromantral folliclesdevelopedinTRL+TcomparedwithTRL alone.DHThad the similar effects to those of high-dose T, except thatDHTreplacement decreased (P <0.05) E2 concentrations produced by fastgrow follicles atWeek 5 regardless of TRL treatment. limitation, reasons for caution: This study reportsTandDHTactionsonin vitro-developedindividualprimate(macaque) follicles, which are limited to the interval from the secondary to small antral stage. wider implication of the findings: 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. study funding/competing interest(s): NIHU54RR024347/RL1HD058294/PL1EB008542(OncofertilityConsortium),NIH U54 HD071836 (SCCPIR), NIHORWH/NICHD 2K12HD043488 (BIRCWH),NIH FICTW/HD-00668, ONPRC 8P51OD011092. There are no conflicts of interest.