Serum levels of anti-müllerian hormone and the ovarian antral follicle count are correlated with ovarian non-growing follicle number

Abstract

OBJECTIVE: Markers of ovarian reserve including serum levels of FSH, Inhibin B, Anti-Müllerian Hormone (AMH), estradiol, and the ovarian antral follicle count as obtained by transvaginal ultrasound examination (AFC) have been correlated with stimulation quality in IVF cycles and with chronological age. This study investigated the relationship between these markers of ovarian reserve and the ovarian non-growing follicle (NGF) number. DESIGN: Prospective study, university setting. MATERIALS AND METHODS: Normal ovaries were collected from 33 women (age 26-52 years) undergoing oophorectomy for benign indications. Prior to surgery each subject underwent a transvaginal ultrasound examination for determination of ovarian AFC and venipuncture for the determination of serum levels of FSH, AMH, Inhibin B and estradiol. After fixation, each ovary was cut into 1 mm “slabs.” Using systematic random sampling rules, eight slabs were selected. The slabs were embedded in glycomethacrylate blocks and exhaustively sectioned at a thickness of 25 μm. Every 10th section was collected, stained, and mounted. A single section representing each slab was selected for counting. The total number of oocyte nucleoli encountered in this fraction was then multiplied by the inverse of a hierarchy of systematic random sampling fractions in order to generate an estimate of the total NGF number in the original specimen. Serum levels of FSH, AMH, Inhibin B and estradiol were determined by ELISA or chemiluminescent assays. Univariate and partial correlations were calculated to determine the association between markers of ovarian reserve and the log of the ovarian NGF number. A p-value of less than 0.05 was considered statistically significant. RESULTS: There were significant correlations between the ovarian NGF count and AFC (r = 0.77, p < 0.0001), AMH (r = 0.70, p < 0.0001), Inhibin B (r = 0.49, p < 0.01), FSH (r = -0.54, p < 0.005) and chronological age (r = -0.80, p < 0.0001). After adjusting for age, significant correlations were identified between the ovarian NGF count and AFC (r = 0.30, p < 0.01) and AMH (r = 0.29, p < 0.01). CONCLUSIONS: Serum levels of AMH and the ovarian AFC are directly correlated with the total ovarian NGF number even after adjustment for chronological age. These observations support the hypothesis that these markers of ovarian reserve are reflective of the primordial follicle pool and provide continued support for their use in the development of models that accurately assess reproductive age.