Anti-Müllerian hormone induces a reversible quiescence-associated secretory phenotype in preantral follicles†

Anti-Müllerian hormone is a factor secreted by granulosa cells of growing follicles that regulates many aspects of ovarian function, including primordial follicle activation and early follicle development through inhibitory feedback. Treatment with exogenous anti-Müllerian hormone at supraphysiological levels can inhibit follicular development and prevent ovulation in mice, rats, and cats. Single-cell transcriptomic analysis of ovaries from mice treated postnatally with anti-Müllerian hormone identified a distinct quiescence signature in granulosa cells. Herein, we further characterized the anti-Müllerian hormone -induced quiescent cell state and drew parallels to developmental senescence by describing a reversible "quiescence-associated secretory phenotype." This state was characterized by the induction of several hallmarks of senescence, including inhibition of proliferation (reduced KI67), upregulation of markers of senescence (Cdkn1a, Fn1, Cebpb, Timp3), and chemokines and their receptors (Cxcl14, Cxcl12, Ccl12, Ccl21, Cxcr2). However, quiescence-associated secretory phenotype did not recapitulate other senescence hallmarks, such as activation of beta-galactosidase activity or permanent exit from the cell cycle. Similar to classical senescence, we found that anti-Müllerian hormone induced quiescence-associated secretory phenotype, in part, through the upregulation of cyclin-dependent kinase inhibitors (Cdkn1a, Cdkn1b, and Cdkn1c) in granulosa cells and oocytes of mouse and human ovaries. Finally, we showed that the quiescence-associated secretory phenotype state could be reversed within a month of discontinuing anti-Müllerian hormone treatment. This recovery was marked by the return of growing primary, secondary, and antral follicles. In conclusion, these data suggest that anti-Müllerian hormone can pause follicle development by inducing quiescence-associated secretory phenotype, a reversible senescent-like state, which may benefit fertility preservation.