Regulation of AMH by oocyte-specific growth factors in human primary cumulus cells

Abstract

The regulation of AMH production by follicular cells is poorly understood. The purpose of this study was to determine the role of the oocyte-secreted factors, growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), on AMH production in primary human cumulus cells. Cumulus cells from IVF patients were cultured with a combination of GDF9, BMP15, recombinant FSH, and specific signaling inhibitors. Stimulation with GDF9 or BMP15 separately had no significant effect on AMH mRNA levels. In contrast, simultaneous stimulation with GDF9 and BMP15 (G+B) resulted in a significant increase in AMH mRNA expression. Increasing concentration of G+B (0.6, 2.5, 5 and 10 ng/ml) stimulated AMH in a dose-dependent manner, showing a maximal effect at 5 ng/ml. Western blot analyses revealed an average 16-fold increase in AMH protein levels in cells treated with G+B when compared to controls. FSH co-treatment decreased the stimulation of AMH expression by G+B. The stimulatory effect of G+B on the expression of AMH was significantly decreased by inhibitors of the SMAD2/3 signaling pathway. These findings show for the first time that AMH production is regulated by oocyte-secreted factors in primary human cumulus cells. Moreover, our novel findings establish that the combination of GDF9 + BMP15 potently stimulates AMH expression.

A novel, noncoding-RNA-mediated, post-transcriptional mechanism of anti-Mullerian hormone regulation by the H19/let-7 axis†

“…AMH is also regulated by oocyte-secreted factors including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15, which synergistically induce AMH expression in cultured human cumulus cells; this stimulatory effect is blunted in the presence of FSH. Inhibition of the SMAD3 pathway, which is involved in GDF9 signaling, also inhibited these effects (Convissar et al 2017) [6]. Thus, AMH may act as a modulator of ovarian steroidogenesis as well as a "gatekeeper" for primordial follicle recruitment.…”

Human follicular fluid shows diverse metabolic profiles at different follicle developmental stages

“…In women, AMH is secreted by ovarian granulosa cells and is observed in pre-antral and small antral follicles. The level of AMH and AFC were consistent, and could reflect the ovarian reserve [22,23]. When antral follicles develop and become larger, AMH expression decreases gradually [24].…”

Plasma pentraxin 3 is higher in early ovarian hyperstimulation syndrome than in uncomplicated in vitro fertilization cycle of high-risk women

“…Circulating PTX3 and AMH levels showed notably different profiles during the IVF cycle, although both are secreted by granulosa cells and regulated by the same oocyte derived factors [25]. Several explanations for this finding may exist.…”

“…Both PTX3 and AMH are secreted from the ovarian granulosa cells: AMH from small antral follicles [22] and PTX3 from antral and periovulatory follicles [12,23]. Moreover, the same oocyte-derived factors, namely growth differentiation factor-9 (GDF9) and bone morphogenic proteins (BMP) [24][25][26] regulate the synthesis of PTX3 and AMH. During the natural menstrual cycle, circulating PTX3 levels are higher during the early follicular (menstruation) than the luteal phase [27].…”