title: 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.
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.
authors.affiliation: Haartman Institute, Department of Bacteriology and Immunology, University of Helsinki, Finland.
authors.forename: R
authors.initials: R
authors.lastname: Jaatinen
authors.forename: V
authors.initials: V
authors.lastname: Rosen
authors.forename: T
authors.initials: T
authors.lastname: Tuuri
authors.forename: O
authors.initials: O
authors.lastname: Ritvos
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D001483:Base Sequence; D055398:Bone Morphogenetic Protein 3; D019485:Bone Morphogenetic Proteins / Q000235:genetics* / Q000378:metabolism*; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000008:administration & dosage / Q000494:pharmacology*; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008184:Luteal Cells / Q000378:metabolism; D010053:Ovary / Q000166:cytology / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: C525757:BMP3 protein, human; D055398:Bone Morphogenetic Protein 3; D019485:Bone Morphogenetic Proteins; D006063:Chorionic Gonadotropin; D017931:DNA Primers; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D015124:8-Bromo Cyclic Adenosine Monophosphate; D013755:Tetradecanoylphorbol Acetate