Gremlin, noggin, chordin and follistatin differentially modulate BMP-induced suppression of androgen secretion by bovine ovarian theca cells

in Journal of Molecular Endocrinology
Correspondence should be addressed to P G Knight: p.g.knight@reading.ac.uk

(M Samir is now at College of Veterinary Medicine, University of Wasit, Wasit, Iraq)

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Bone morphogenetic proteins (BMPs) are firmly implicated as intra-ovarian regulators of follicle function and steroidogenesis, but information is lacking regarding the regulation of BMP signalling by extracellular binding proteins co-expressed in the ovary. In this study, we compared the abilities of four BMP-binding proteins (gremlin, noggin, chordin, follistatin) to antagonize the action of four different BMPs (BMP2 BMP4, BMP6, BMP7) on LH-induced androstenedione secretion by bovine theca cells in primary culture. Expression of the four BMP-binding proteins and BMPs investigated here has previously been documented in bovine follicles. All four BMPs suppressed androstenedione secretion by >85%. Co-treatment with gremlin antagonized BMP2- and, less potently, BMP4-induced suppression of androgen secretion but did not affect responses to BMP6 and BMP7. Noggin antagonized the effects of three BMPs (rank order: BMP4 > BMP2 > BMP7) but did not affect the response to BMP6. Follistatin partially reversed the suppressive effects of BMP6 on androgen secretion but did not affect BMP2, BMP4 and BMP7 action. Chordin had no effect on the response to any of the four BMPs. BMP6 treatment upregulated thecal expression of GREM1, NOG, CHRD and SMAD6 mRNA whilst inhibiting expression of the four BMPs. Taken together with previous work documenting the intra-ovarian expression of different BMPs, BMP-binding proteins and signalling receptors, these observations reinforce the conclusion that extracellular binding proteins selectively modulate BMP-dependent alterations in thecal steroidogenesis. As such they likely constitute an important regulatory component of this and other intra-ovarian actions of BMPs.

 

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    Effects of (A) LH and (B) BMP2, BMP4, BMP6 and BMP7 on secretion of androstenedione and progesterone by bovine theca interna cells and on viable cell number at the end of culture. In (B) cells were cultured in the presence of LH. Values are means and bars indicate SEM (n = 3 independent cultures). ***P < 0.001 vs control.

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    Effects of gremlin on secretion of (A) androstenedione and (B) progesterone by bovine theca interna cells treated with BMP2, BMP4, BMP6 or BMP7 under LH-stimulated conditions. Values are means and bars indicate s.e.m. (n = 3 independent experiments). Results of 2-way ANOVA are indicated. Within each BMP treatment group, means without a common letter are significantly (P < 0.05) different.

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    Effects of noggin on secretion of (A) androstenedione and (B) progesterone by bovine theca interna cells treated with BMP2, BMP4, BMP6 or BMP7 under LH-stimulated conditions. Values are means and bars indicate s.e.m. (n = 3 independent experiments). Results of 2-way ANOVA are indicated. Within each BMP treatment group, means without a common letter are significantly (P < 0.05) different.

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    Effects of follistatin on secretion of (A) androstenedione and (B) progesterone by bovine theca interna cells treated with BMP2, BMP4, BMP6 or BMP7 under LH-stimulated conditions. Values are means and bars indicate s.e.m. (n = 3 independent experiments). Results of 2-way ANOVA are indicated. Within each BMP treatment group, means without a common letter are significantly (P < 0.05) different.

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    Effects of chordin on secretion of (A) androstenedione and (B) progesterone by bovine theca interna cells treated with BMP2, BMP4, BMP6 or BMP7 under LH-stimulated conditions. Values are means and bars indicate s.e.m. (n = 3 independent experiments). Results of 2-way ANOVA are indicated. Within each BMP treatment group, means without a common letter are significantly (P < 0.05) different.

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    Time-dependent effect of BMP6 treatment on relative abundance of transcripts for (A) GREM1, (B) CHRD, (C) NOG, (D) FST, (E) BMP2, (F) BMP4, (G) BMP6, (H) BMP7 and (I) SMAD6 in cultured bovine theca interna cells. Values are means and bars indicate s.e.m. (n = 4 independent experiments). *P < 0.05, **P < 0.01, ***P < 0.001 vs control.

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