Transforming growth factor beta1 regulates follistatin mRNA expression during in vitro bovine granulosa cell differentiation

J Cell Physiol. 2006 Apr;207(1):40-8. doi: 10.1002/jcp.20533.


In order to test the hypothesis that transforming growth factor beta (TGF-beta) acts by FS regulation on bovine granulosa cells in in vitro differentiation, we analyzed the effect of TGF-beta1 on follistatin mRNA expression in three differentiation states of bovine granulosa cells. We showed a positive regulation of FS mRNA after TGF-beta1 (1 ng/ml) treatment of freshly isolated granulosa cells from small-medium antral follicles (2-8 mm). This effect was abolished by the addition of exogenous follistatin (100 ng/ml), suggesting that this effect could be mediated by activin. Although these cells showed a similar effect on FS mRNA expression after treatment with activin-A, a soluble form of activin receptor type IIA was unable to inactivate the TGF-beta effect. When we tested the TGF-beta effect on FS mRNA in different granulosa cell states, TGF-beta1 regulation was associated with progesterone production only in freshly isolated cells. The amount of total activin-A produced by first passage cells (dedifferentiated cells), was ten times smaller than the one measured in a conditioned medium from freshly isolated cells (mature cells). The TGF-beta1-dependent FS mRNA expression persisted in first passage cells without changes with FS addition. On the other hand, the BGC-1 granulosa cell line (immature cells) produced large amounts of activin-A regulated by TGF-beta1 and an invariable steady state of FS mRNAs. In summary, our results showed that FS mRNA expression is regulated by TGF-beta1 independently of activin effects in differentiated granulosa cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activin Receptors, Type II / pharmacology
  • Activins / metabolism
  • Activins / pharmacology
  • Animals
  • Binding, Competitive
  • Cattle
  • Cell Differentiation / genetics*
  • Cell Line
  • Cells, Cultured
  • Culture Media, Conditioned / chemistry
  • Female
  • Fibronectins / metabolism
  • Follistatin / genetics*
  • Follistatin / pharmacology
  • Gene Expression / drug effects
  • Granulosa Cells / cytology
  • Granulosa Cells / drug effects*
  • Granulosa Cells / metabolism
  • Humans
  • Inhibin-beta Subunits / metabolism
  • Inhibin-beta Subunits / pharmacology
  • Progesterone / metabolism
  • Protein-Serine-Threonine Kinases
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Receptor, Transforming Growth Factor-beta Type II
  • Receptors, Transforming Growth Factor beta / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transforming Growth Factor beta / metabolism
  • Transforming Growth Factor beta / pharmacology*
  • Transforming Growth Factor beta1


  • Culture Media, Conditioned
  • Fibronectins
  • Follistatin
  • RNA, Messenger
  • Receptors, Transforming Growth Factor beta
  • TGFB1 protein, human
  • Transforming Growth Factor beta
  • Transforming Growth Factor beta1
  • activin A
  • Activins
  • Progesterone
  • Inhibin-beta Subunits
  • Protein-Serine-Threonine Kinases
  • Activin Receptors, Type II
  • Receptor, Transforming Growth Factor-beta Type II
  • activin receptor type II-A