Single-cell sequencing reveals suppressive transcriptional programs regulated by MIS/AMH in neonatal ovaries

Proc Natl Acad Sci U S A. 2021 May 18;118(20):e2100920118. doi: 10.1073/pnas.2100920118.


Müllerian inhibiting substance (MIS/AMH), produced by granulosa cells of growing follicles, is an important regulator of folliculogenesis and follicle development. Treatment with exogenous MIS in mice suppresses follicle development and prevents ovulation. To investigate the mechanisms by which MIS inhibits follicle development, we performed single-cell RNA sequencing of whole neonatal ovaries treated with MIS at birth and analyzed at postnatal day 6, coinciding with the first wave of follicle growth. We identified distinct transcriptional signatures associated with MIS responses in the ovarian cell types. MIS treatment inhibited proliferation in granulosa, surface epithelial, and stromal cell types of the ovary and elicited a unique signature of quiescence in granulosa cells. In addition to decreasing the number of growing preantral follicles, we found that MIS treatment uncoupled the maturation of germ cells and granulosa cells. In conclusion, MIS suppressed neonatal follicle development by inhibiting proliferation, imposing a quiescent cell state, and preventing granulosa cell differentiation.

Keywords: Müllerian inhibiting substance; granulosa cells; ovarian reserve; ovary; scRNA-seq.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Anti-Mullerian Hormone / pharmacology*
  • Cell Differentiation / drug effects
  • Female
  • Inhibins / analysis
  • Mice
  • Mice, Inbred C57BL
  • Ovarian Follicle / drug effects
  • Ovarian Follicle / physiology
  • Ovary / drug effects*
  • Ovary / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Peptide / analysis
  • Receptors, Transforming Growth Factor beta / analysis
  • Sequence Analysis, RNA
  • Single-Cell Analysis
  • Transcription, Genetic / drug effects


  • Receptors, Peptide
  • Receptors, Transforming Growth Factor beta
  • anti-Mullerian hormone receptor
  • inhibin-alpha subunit
  • Inhibins
  • Anti-Mullerian Hormone