Identification of ovarian granulosa cells as a novel site of expression for bone morphogenetic protein-3 (BMP-3/osteogenin) and regulation of BMP-3 messenger ribonucleic acids by chorionic gonadotropin in cultured human granulosa-luteal cells

J Clin Endocrinol Metab. 1996 Nov;81(11):3877-82. doi: 10.1210/jcem.81.11.8923832.

Abstract

Bone morphogenetic proteins (BMP) belong structurally to the transforming growth factor-beta superfamily comprising several growth and differentiation factors such as inhibin, activin, and Müllerian inhibitory factor that regulate ovarian function. We studied here the potential expression of BMP-2, -3, and -4 messenger RNAs (mRNAs) in isolated human granulosa cells obtained at oocyte retrieval for in vitro fertilization. Freshly isolated granulosa cells were found to express BMP-3 (also known as osteogenin) mRNAs but not those of BMP-2 or -4. The BMP-3 transcripts were detected with RT-PCR amplification followed by Southern blot hybridization, as well as by Northern and dot blot hybridization analyses. To investigate whether BMP-3 mRNAs are hormonally regulated, cultures of human granulosa-luteal (GL) cells were treated with different concentrations of purified human chorionic gonadotropin (hCG) at varying stages of culture. hCG decreased BMP-3 mRNA levels from the first day of the culture up to day 5. Time-dependence studies showed that a clear decrease in BMP-3 mRNA levels was evident at 24 h after hCG treatment, and that the effect of hCG was concentration dependent with 3 ng/mL hCG decreasing significantly (P < 0.05) BMP-3 mRNA levels. Furthermore, the cAMP analog, 8-bromo-cAMP (8-Br-cAMP), which activates protein kinase-A, and 12-0-tetradecanoylphorbol 13-acetate, an activator of protein kinase-C, both markedly decreased BMP-3 mRNA levels in an 8-h treatment. We conclude that: 1) BMP-3 mRNAs are expressed in human granulosa cells; 2) their steady state levels are hormonally regulated in cultured human GL cells as evidenced by the ability of hCG to markedly decrease BMP-3 transcript levels; and (3) that activation of both protein kinase-A-and protein kinase-C-mediated signaling pathways also results in a decrease in BMP-3 mRNA levels in GL cells. We suggest that BMP-3, like several other members of the transforming growth factor-beta superfamily, is a potential local regulator of female gonadal function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 8-Bromo Cyclic Adenosine Monophosphate / pharmacology
  • Base Sequence
  • Bone Morphogenetic Protein 3
  • Bone Morphogenetic Proteins / genetics*
  • Bone Morphogenetic Proteins / metabolism*
  • Cells, Cultured
  • Chorionic Gonadotropin / administration & dosage
  • Chorionic Gonadotropin / pharmacology*
  • DNA Primers / genetics
  • Female
  • Gene Expression Regulation / drug effects
  • Granulosa Cells / metabolism*
  • Humans
  • In Vitro Techniques
  • Kinetics
  • Luteal Cells / metabolism
  • Ovary / cytology
  • Ovary / metabolism
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Transforming Growth Factor beta / metabolism

Substances

  • BMP3 protein, human
  • Bone Morphogenetic Protein 3
  • Bone Morphogenetic Proteins
  • Chorionic Gonadotropin
  • DNA Primers
  • RNA, Messenger
  • Transforming Growth Factor beta
  • 8-Bromo Cyclic Adenosine Monophosphate
  • Tetradecanoylphorbol Acetate