Journal
JOURNAL OF ASSISTED REPRODUCTION AND GENETICS
Volume 33, Issue 12, Pages 1665-1675Publisher
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s10815-016-0804-3
Keywords
Anti-Mullerian hormone; Three-dimensional follicle culture; RNA sequencing; Preantral follicle; Antral follicle
Funding
- National Institutes of Health (NIH) Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD) [R01HD082208]
- NIH Office of Research on Women's Health/NICHD (Building Interdisciplinary Research Careers in Women's Health, BIRCWH) [K12HD043488]
- NIH Office of the Director ONPRC Pilot Grant [P51OD011092]
- Collins Medical Trust
- American Society for Reproductive Medicine
- Medical Research Foundation of Oregon
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The main goals of this study were to investigate the expression of anti-Mullerian hormone (AMH) and its receptor (AMHR2) during follicular development in primates, and to evaluate the potential of AMH as a biomarker for follicle growth and oocyte maturation in vitro. The mRNA and protein expression of AMH and AMHR2 were determined using isolated follicles and ovarian sections from rhesus macaques (n = 4) by real-time PCR and immunohistochemistry, respectively. Isolated secondary follicles were cultured individually. Follicle growth and media AMH concentrations were assessed by ELISA. The mRNA expression profiles, obtained from RNA sequencing, of in vitro- and in vivo-developed antral follicles were compared. Secondary follicles from additional animals (n = 35) were cultured. Follicle growth, oocyte maturation, and media AMH concentrations were evaluated for forecasting follicular development in vitro by AMH levels. AMH immunostaining was heterogeneous in the population of preantral follicles that were also stained for AMHR2. The mRNA expression profiles were comparable between in vivo- and in vitro-developed follicles. AMH levels produced by growing follicles were higher than those of nongrowing follicles in culture. With a cutoff value of 1.40 ng/ml, 85 % of nongrowing follicles could be identified while eliminating only 5 % of growing follicles. Growing follicles that generated metaphase II-stage oocytes secreted greater amounts of AMH than did those yielding immature germinal vesicle-stage oocytes. AMH, co-expressed with AMHR2, was produced heterogeneously by preantral follicles in macaques with levels correlated positively with follicle growth and oocyte maturation. AMH may serve as a biomarker for primate follicular development in vitro.
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