Anti-Mullerian hormone (AMH) protects ovarian follicle loss by downregulating granulosa cell function in in vitro and in vivo models.

Purpose AMH inhibits hormone production in luteinized granulosa cells (GCs) and stalls ovarian follicle development in vitro and in vivo. We sought to confirm AMH’s mechanism of action through SMAD activation and investigate AMH inhibition of follicle development and function, in vitro and in vivo.

A primary culture of GCs isolated from follicular fluid was used, and cells were treated with recombinant AMH (rAMH) or placebo for 24 h. For the mouse model, 18-weeks old C57BL female mice were either euthanized at the beginning or treated with rAMH or normal saline for 3 weeks. Primordial (PDF), primary follicle (PRF), secondary (SEF), and tertiary follicles (TEF) were calculated. Real-time RT-PCR and ELISA were performed to quantify GC gene expression and protein translation of human SMAD 1, 5, and 8, FSH-R and mouse FSH-R, inhibin B, caspase 3, Ki67, BMP15, GDF9, and the epigenetic regulators miRNAa and b.

In vitro, rAMH-treated GC showed activation of the SMAD 1, 5 and downregulation of SMAD 8, with greater magnitude at increasing rAMH doses (p < 0.04) and consequential control of downstream regulators. In vivo, the rAMH-treated mice showed increased SEFs and decreased PRFs while PDFs, TEFs, were unchanged compared with baseline. Compared with Placebo, the rAMH group showed increased PDFs, while PRFs, and TEFs were significantly decreased, and SEFs were unchanged.

AMH caused SMAD activation in a dose-dependent manner, with downstream downregulation of cell function and replication, also through activation of miRNAs. These mechanisms were confirmed by the in vivo findings with ultimate downregulation of follicular development and preservation of the ovarian follicle number. Counteracting follicular depletion, AMH could be used to protect the ovarian follicle reservoir. Similar content being viewed by others Data availability The data described in this manuscript is kept in a private database and can be made available to the reviewers.

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Author information Authors and Affiliations Contributions LD and GMS conceived and designed research for the reported studies. LD and GMS conducted in vivo and in vitro experiments. GMS contributed all reagents and analytical tools. LD and MCM analyzed data and produced all tables and figures. LD, MCM, MPD, and GMS wrote the manuscript. All authors read and approved the manuscript. Corresponding author Ethics declarations Ethical approval The different studies here described were approved by an internal ethics review board at IVF Michigan Rochester Hills and Flint, PC, in 2017 and by the University of Tennessee Health Science Center (UTHSC) Institutional Animal Care and Use Committee (IACUC) in 2015. Conflict of interest The authors declare no competing interests. Additional information Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Rights and permissions Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. About this article Cite this article Detti, L., Mari, M.C., Diamond, M.P. et al. Anti-Mullerian hormone (AMH) protects ovarian follicle loss by downregulating granulosa cell function in in vitro and in vivo models. J Assist Reprod Genet 42, 2029–2037 (2025). https://doi.org/10.1007/s10815-025-03473-x Received: Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s10815-025-03473-x