Growth differentiation factor 3 is induced by bone morphogenetic protein 6 (BMP-6) and BMP-7 and increases luteinizing hormone receptor messenger RNA expression in human granulosa cells.
OBJECTIVE
To examine the relevance of growth differentiation factor 3 (GDF-3) and bone morphogenetic protein (BMP) cytokines in human ovary.


METHODS
Molecular studies.


METHODS
Research laboratory.


METHODS
Eight women undergoing salpingo-oophorectomy and 30 women undergoing ovarian stimulation for in vitro fertilization.


METHODS
Localizing GDF-3 protein in human ovaries; granulosa cells (GC) cultured with GDF-3, BMP-6, or BMP-7 followed by RNA extraction.


METHODS
The localization of GDF-3 protein in normal human ovaries via immunohistochemical analysis, GDF-3 messenger RNA (mRNA) expression evaluation via quantitative real-time reverse transcription and polymerase chain reaction (RT-PCR), and evaluation of the effect of GDF-3 on leuteinizing hormone (LH) receptor mRNA expression via quantitative real-time RT-PCR.


RESULTS
In the ovary, BMP cytokines, of the transforming growth factor beta (TGF-β) superfamily, are known as a luteinization inhibitor by suppressing LH receptor expression in GC. Growth differentiation factor 3, a TGF-β superfamily cytokine, is recognized as an inhibitor of BMP cytokines in other cells. Immunohistochemical analysis showed that GDF-3 was strongly detected in the GC of antral follicles. An in vitro assay revealed that BMP-6 or BMP-7 induced GDF-3 mRNA in GC. Also, GDF-3 increased LH receptor mRNA expression and inhibited the effect of BMP-7, which suppressed the LH receptor mRNA expression in GC.


CONCLUSIONS
GDF-3, induced with BMP-6 and BMP-7, might play a role in folliculogenesis by inhibiting the effect of BMP cytokines.
Department of Obstetrics and Gynecology, University of Tokyo, Tokyo, Japan.
Jia
J
Shi
Osamu
O
Yoshino
Yutaka
Y
Osuga
Ikumi
I
Akiyama
Miyuki
M
Harada
Kaori
K
Koga
Akihisa
A
Fujimoto
Tetsu
T
Yano
Yuji
Y
Taketani
D000328:Adult; D055418:Bone Morphogenetic Protein 6 / Q000378:metabolism*; D055419:Bone Morphogenetic Protein 7 / Q000378:metabolism*; D002478:Cells, Cultured; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D055451:Growth Differentiation Factor 3 / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007986:Luteinizing Hormone / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D011974:Receptors, LH / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
C525816:BMP6 protein, human; C525819:BMP7 protein, human; D055418:Bone Morphogenetic Protein 6; D055419:Bone Morphogenetic Protein 7; C525974:GDF3 protein, human; D055451:Growth Differentiation Factor 3; D012333:RNA, Messenger; D011974:Receptors, LH; D007986:Luteinizing Hormone