title: Which follicles make the most anti-Mullerian hormone in humans? Evidence for an abrupt decline in AMH production at the time of follicle selection.
abstract: Anti-Müllerian hormone (AMH) is exclusively produced by granulosa cells (GC) of the developing pre-antral and antral follicles, and AMH is increasingly used to assess ovarian function. It is unclear which size follicles make the most AMH (total content) and are the main contributors to circulating AMH concentrations. To determine AMH gene expression in GC (q-RT-PCR) and follicular AMH production (Elisa and RIA) in relation to follicular development, 87 follicles (3-13 mm diameter) including both GC and the corresponding follicular fluid (FF) were collected in connection with fertility preservation of human ovaries. Further, follicle number and diameter, graded in 1 mm increments, were determined by 3D ultrasound in 113 women in their natural menstrual cycle to determine follicle number and diameter in relation to circulating AMH levels. This study demonstrates for the first time a positive association between AMH gene expression in human and both total follicular fluid AMH (P < 0.02) and follicular fluid AMH concentration (P < 0.01). AMH gene expression and total AMH protein increased until a follicular diameter of 8 mm, after which a sharp decline occurred. In vivo modelling confirmed that 5-8 mm follicles make the greatest contribution to serum AMH, estimated for the first time in human to be 60% of the circulating concentration. Significant positive associations between gene expression of AMH and FSHR, AR and AMHR2 expression (P < 0.00001 for all three) and significant negative association between follicular fluid AMH concentration and CYP19a1 expression were found (P < 0.0001). Both AMH gene expression (P < 0.02) and follicular fluid concentration of AMH (P < 0.00001) correlated negatively with estradiol concentration.
authors.affiliation: Laboratory of Reproductive Biology, Section 5712, The Juliane Marie Centre for Women, Children and Reproduction, University Hospital of Copenhagen, University of Copenhagen, Blegdamsvej 9, Rigshospitalet, DK-2100 Copenhagen, Denmark.
authors.forename: J V
authors.initials: JV
authors.lastname: Jeppesen
authors.forename: R A
authors.initials: RA
authors.lastname: Anderson
authors.forename: T W
authors.initials: TW
authors.lastname: Kelsey
authors.forename: S L
authors.initials: SL
authors.lastname: Christiansen
authors.forename: S G
authors.initials: SG
authors.lastname: Kristensen
authors.forename: K
authors.initials: K
authors.lastname: Jayaprakasan
authors.forename: N
authors.initials: N
authors.lastname: Raine-Fenning
authors.forename: B K
authors.initials: BK
authors.lastname: Campbell
authors.forename: C
authors.initials: C
authors.lastname: Yding Andersen
mesh_terms: D000293:Adolescent; D000328:Adult; D054304:Anti-Mullerian Hormone / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D001141:Aromatase / Q000096:biosynthesis; D002648:Child; D004958:Estradiol / Q000097:blood; D005260:Female; D015571:Follicular Fluid / Q000378:metabolism*; D015870:Gene Expression; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D011962:Receptors, FSH / Q000096:biosynthesis; D018000:Receptors, Peptide / Q000096:biosynthesis; D018125:Receptors, Transforming Growth Factor beta / Q000096:biosynthesis; D055815:Young Adult
chemical_list: D011962:Receptors, FSH; D018000:Receptors, Peptide; D018125:Receptors, Transforming Growth Factor beta; C084621:anti-Mullerian hormone receptor; D004958:Estradiol; D054304:Anti-Mullerian Hormone; D001141:Aromatase; C561410:CYP19A1 protein, human
keywords: AMH; follicular fluid; granulosa cells; human small antral follicles; modelling