title: The anti-Müllerian hormone (AMH) acts as a gatekeeper of ovarian steroidogenesis inhibiting the granulosa cell response to both FSH and LH.
abstract: OBJECTIVE
  Anti Müllerian Hormone (AMH) has a negative and inhibitory role in many functions of human granulosa-lutein cells (hGCs) including notoriously the reduction of the aromatase CYP19A1 expression induced by follicle-stimulating hormone (FSH). No data have been provided on the possible role of AMH in modulating the response to luteinizing hormone (LH) (alone or combined with FSH) as well as its effect on other enzymes involved in steroidogenesis including aromatase P450scc. The aim of this study was to investigate the role of AMH as regulator of the basal and stimulated steroids production by hGCs.
  
  
  METHODS
  Primary culture of hGCs were incubated with hormones AMH, LH, and FSH, alone or in combination. The CYP19A1 and P450scc messenger RNA (mRNA) expression, normalized by housekeeping ribosomal protein S7 (RpS7) gene, was evaluated by reverse transcriptase quantitative PCR (RT-qPCR). Each reaction was repeated in triplicate. Negative controls using corresponding amount of vehicle control for each hormone treatment were performed.
  
  
  RESULTS
  AMH did not modulate the basal mRNA expression of both aromatase genes at any of the concentrations tested. Meanwhile, the strong mRNA induction of CYP19A1 and P450scc generated by a 24-h gonadotropin treatment (alone and combined) was suppressed by 20 ng/ml AMH added to culture medium.
  
  
  CONCLUSIONS
  These findings contribute in clarifying the relationship between hormones regulating the early phase of steroidogenesis confirming that AMH is playing a suppressive role on CYP19A1 expression stimulated by gonadotropin in hGCs. Furthermore, a similar inhibitory effect for AMH was observed on P450scc gene expression when activated by gonadotropin treatment.
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy.
authors.forename: Sandro
authors.initials: S
authors.lastname: Sacchi
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: D'Ippolito
authors.affiliation: Department of Biomedical, Metabolic and Neural Sciences, Section of Human Morphology, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Sena
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy.
authors.forename: Tiziana
authors.initials: T
authors.lastname: Marsella
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Tagliasacchi
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Maggi
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy.
authors.forename: Cindy
authors.initials: C
authors.lastname: Argento
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Tirelli
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Giulini
authors.affiliation: Mother-Infant Department, Institute of Obstetrics and Gynecology, University of Modena and Reggio Emilia, Policlinico di Modena, 41100, Modena, Italy. antlamarca@libero.it.
authors.forename: Antonio
authors.initials: A
authors.lastname: La Marca
mesh_terms: D054304:Anti-Mullerian Hormone / Q000008:administration & dosage / Q000378:metabolism*; D001141:Aromatase / Q000096:biosynthesis*; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000096:biosynthesis*; D003470:Culture Media / Q000737:chemistry; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D005640:Follicle Stimulating Hormone / Q000008:administration & dosage / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006062:Gonadotropins / Q000008:administration & dosage; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007986:Luteinizing Hormone / Q000008:administration & dosage / Q000378:metabolism; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000096:biosynthesis
chemical_list: D003470:Culture Media; D006062:Gonadotropins; D012333:RNA, Messenger; D004958:Estradiol; D054304:Anti-Mullerian Hormone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D001141:Aromatase; C561410:CYP19A1 protein, human; D002786:Cholesterol Side-Chain Cleavage Enzyme
keywords: AMH; Aromatase; Ovarian enzyme; P450scc; Steroidogenesis