Attenuation of Bone Morphogenetic Proteins (BMPs) Signaling Induces Granulosa Cell Sensitivity to Gonadotropins in Female Mouse

Jump To References Section


  • Laboratory of Human Growth and Reproductive Development, Department of Pediatrics, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, New Brunswick 08901, NJ ,US
  • School of Science and Technology, University of New England, Armidale 2351, NSW ,AU
  • School of Science and Technology, University of New England, Armidale 2351, NSW ,AU
  • Curtin Medical School, Curtin Health Innovation Research Institute (CHIRI), Curtin University, Bentley 6012, WA ,AU
  • School of Science and Technology, University of New England, Armidale 2351, NSW ,AU
  • Curtin Medical School, Curtin Health Innovation Research Institute (CHIRI), Curtin University, Bentley 6012, WA ,AU



BMP-4, BMPR1B, FSHR, Gene Expression, Immunization, Immunofluorescence, LHCGR, Protein Expression


Several lines of evidence support the notion that Bone Morphogenetic Proteins (BMPs) and gonadotropic hormones are major regulators of ovarian follicle development by inducing gonadotropin-mediated steroid hormone production and regulating ovarian follicle cell proliferation. Research from our laboratory and elsewhere have demonstrated that BMPs play important roles during the early stages of folliculogenesis. Despite the extensive research to reveal the mechanism of interaction between BMPs and gonadotropic hormones in the process of folliculogenesis, the exact mechanism of such an interaction is not adequately understood. Previously, we developed a unique mouse model characterized by a short-term attenuation of the BMP signaling system using passive immunization against bone morphogenetic protein 4 (BMP-4) and bone morphogenetic protein receptor 1B (BMPR1B). This model unequivocally demonstrated that the attenuation of BMP action was an effective method of enhancing fertility reserve and promoting follicle and ovulation rates in female mice. In addition, this study pointed to the possibility of bi-directional mutual regulation between BMPs, Follicle-Stimulating Hormone (FSH), and Luteinizing Hormone (LH). To gain further insight into this mechanism we used this mouse model to examine the protein expression and mRNA level of BMPR1B, Follicle-Stimulating Hormone Receptor (FSHR) and Luteinizing Hormone/Choriogonadotropin Receptor (LHCGR) in multiple stages of follicle development in female mice. Immunofluorescent analysis of female mice treated with anti-BMPR1B antibodies showed a significant upregulation of BMPR1B, FSHR, and LHCGR in the ovarian granulosa cells during the main stages of follicle development, whereas treatment with anti-BMP4 antibodies showed no effect. In addition, attenuation of BMPR1B resulted in upregulation of the FSHR (exclusively expressed in the granulosa cell) and LHCGR mRNA levels in the ovary. The present study implies that BMPs engage indirectly in regulating the later stages of folliculogenesis, in addition to their direct role in the regulation of the early stages of follicle development, by enhancing granulosa cell sensitivity to gonadotropins through upregulating the receptor expression. We propose that our mouse model is siutable to elucidate the mechanism of interaction between BMPs and gonadotropins in folliculogenesis.


Download data is not yet available.


Metrics Loading ...




How to Cite

Al-Samerria, S., Palanisamy, S. K. A., Al-Ali, I., Mamotte, C. D., McFarlane, J. R., & Almahbobi, G. (2022). Attenuation of Bone Morphogenetic Proteins (BMPs) Signaling Induces Granulosa Cell Sensitivity to Gonadotropins in Female Mouse. Journal of Endocrinology and Reproduction, 26(2), 107–118.



Original Research



McGee EA, Hsueh AJ. Initial and cyclic recruitment of ovarian follicles. Endocrine Reviews. 2000; 21(2):200–14. PMid:10782364

Grieve KJ, Freiman RN. The developmental origins of the mammalian ovarian reserve. Development (Cambridge, England). 2015; 142(15):2554–63. PMid:26243868. PMCid:PMC4529035

Rimon-Dahari N, Yerushalmi-Heinemann L, Alyagor L, Dekel N. Ovarian folliculogenesis. Results and problems in cell differentiation. 2016; 58:167–90. PMid:27300179

Uslu B, Dioguardi CC, Haynes M, et al. Quantifying growing versus non-growing ovarian follicles in the mouse. Journal of Ovarian Research. 2017; 10(1):3. PMid:28086947. PMCid:PMC5237173

Huang HJ, Wu JC, Su P, et al. A novel role for bone morphogenetic proteins in the synthesis of follicle-stimulating hormone. Endocrinology. 2001; 142(6):2275–83. PMid:11356673

Takeda M, Otsuka F, Takahashi H, et al. Interaction between gonadotropin-releasing hormone and bone morphogenetic protein-6 and -7 signaling in L?T2 gonadotrope cells. Molecular and Cellular Endocrinology. 2012; 348(1):147–54. PMCid:PMC3719407

Al-Samerria S, Al-Ali I, McFarlane JR, Almahbobi G. The impact of passive immunisation against BMPRIB and BMP4 on follicle development and ovulation in mice. Reproduction (Cambridge, England). 2015; 149(5):403–11. PMid:25667430

Otsuka F. Modulation of bone morphogenetic protein activity by melatonin in ovarian steroidogenesis. Reproductive Medicine and Biology. 2018; 17(3):228–33. PMid:30013422. PMCid:PMC6046534

Shi J, Yoshino O, Osuga Y, et al. Bone morphogenetic protein 7 (BMP-7) increases the expression of Follicle-Stimulating Hormone (FSH) receptor in human granulosa cells. Fertility and Sterility. 2010; 93(4):1273–9. PMid:19108831

Haugen MJ, Johnson AL. Bone morphogenetic protein 2 inhibits FSH responsiveness in hen granulosa cells. Reproduction (Cambridge, England). 2010; 140(4):551–8. PMid:20639315

Faure MO, Nicol L, Fabre S, et al. BMP-4 inhibits follicle-stimulating hormone secretion in ewe pituitary. The Journal of Endocrinology. 2005; 186(1):109–21. PMid:16002541

Lee WS, Otsuka F, Moore RK, Shimasaki S. Effect of bone morphogenetic protein-7 on folliculogenesis and ovulation in the rat. Biology of Reproduction. 2001; 65(4):994–9. PMid:11566718

Lawson KA, Dunn NR, Roelen BA, et al. Bmp4 is required for the generation of primordial germ cells in the mouse embryo. Genes and Development. 1999; 13(4):424–36. PMid:10049358. PMCid:PMC316469

Nilsson EE, Skinner MK. Bone morphogenetic protein-4 acts as an ovarian follicle survival factor and promotes primordial follicle development. Biology of Reproduction. 2003; 69(4):1265–72. PMid:12801979

Knight PG, Glister C. Local roles of TGF-beta superfamily members in the control of ovarian follicle development. Animal Reproduction Science. 2003; 78(3-4):165–83.

Knight PG, Glister C. TGF-beta superfamily members and ovarian follicle development. Reproduction (Cambridge, England). 2006; 132(2):191–206. PMid:16885529

Lee KF, Chow JF, Xu JS, et al. A comparative study of gene expression in murine embryos developed in vivo, cultured in vitro, and cocultured with human oviductal cells using messenger ribonucleic acid differential display. Biology of Reproduction. 2001; 64(3):910–7. PMid:11207208

Sun RZ, Lei L, Cheng L, et al. Expression of GDF-9, BMP-15 and their receptors in mammalian ovary follicles. Journal of Molecular Histology. 2010; 41(6):325–32. PMid:20857181

Shimasaki S, Zachow RJ, Li D, Kim H, et al. A functional bone morphogenetic protein system in the ovary. Proceedings of the National Academy of Sciences of the United States of America. 1999; 96(13):7282–7. PMid:10377406. PMCid:PMC22077

Hosoya T, Otsuka F, Nakamura E, et al. Regulatory role of BMP-9 in steroidogenesis by rat ovarian granulosa cells. The Journal of Steroid Biochemistry and Molecular Biology. 2014; 147C:85–91. PMid:25527306

Yoshino O, Shi J, Osuga Y, et al. The function of bone morphogenetic proteins in the human ovary. Reprod Med Biol. 2011; 10(1):1–7. PMid:29699076. PMCid:PMC5904583

Moore RK, Erickson GF, Shimasaki S. Are BMP-15 and GDF-9 primary determinants of ovulation quota in mammals? Trends in Endocrinology and Metabolism: TEM. 2004; 15(8):356–61. PMid:15380806

Araújo VR, Lima-Verde IB, Name KPO, et al. Bone Morphogenetic Protein-6 (BMP-6) induces atresia in goat primordial follicles cultured in vitro. Pesquisa Veterinária Brasileira. 2010; 30:770–81.

Yang Y, Kanno C, Huang W, K et al. Effect of bone morphogenetic protein-4 on in vitro growth, steroidogenesis and subsequent developmental competence of the oocyte-granulosa cell complex derived from bovine early antral follicles. Reproductive Biology and Endocrinology: RB&E. 2016; 14:3. PMid:26769429. PMCid:PMC4714481

Abel MH, Wootton AN, Wilkins V, et al. The effect of a null mutation in the follicle-stimulating hormone receptor gene on mouse reproduction. Endocrinology. 2000; 141(5):1795–803. PMid:10803590

Shimasaki S, Moore RK, Otsuka F, Erickson GF. The bone morphogenetic protein system in mammalian reproduction. Endocrine Reviews. 2004; 25(1):72–101. PMid:14769828

Ruoss C, Tadros A, O’Shea T, et al. Ovarian follicle development in Booroola sheep exhibiting impaired bone morphogenetic protein signalling pathway. Reproduction (Cambridge, England). 2009; 138(4):689–96. PMid:19589856

Tanwar PS, O’Shea T, McFarlane JR. In vivo evidence of role of bone morphogenetic protein-4 in the mouse ovary. Animal Reproduction Science. 2008; 106(3–4):232–40. PMid:17644284

Tanwar PS, McFarlane JR. Dynamic expression of bone morphogenetic protein 4 in reproductive organs of female mice. Reproduction (Cambridge, England). 2011; 142(4):573–9. PMid:21810858

McKinney MM, Parkinson A. A simple, non-chromatographic procedure to purify immunoglobulins from serum and ascites fluid. Journal of Immunological Methods. 1987; 96(2):271–8.

Al-Samerria S, Almahbobi G. Three-dimensional image analysis to quantify the temporo-spatial expression of cellular receptors. Journal of Medical and Bioengineering. 2014; 3(3):179–82.

Ciller IM, Palanisamy SKA, Ciller UA, McFarlane JR. Postnatal expression of bone morphogenetic proteins and their receptors in the mouse testis. Physiology Research Journal. 2015; In Press. PMid:26988160

Ye J, Coulouris G, Zaretskaya I, Cutcutache I, Rozen S, Madden TL. Primer-BLAST: A tool to design target-specific primers for polymerase chain reaction. BMC Bioinformatics. 2012; 13(1):134. PMid:22708584. PMCid:PMC3412702

Pedersen T, Peters H. Follicle growth and cell dynamics in the mouse ovary during pregnancy. Fertility and Sterility. 1971; 22(1):42–52.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods (San Diego, Calif). 2001; 25(4):402–8. PMid:11846609

Vandesompele J, De Preter K, Pattyn F, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology. 2002; 3(7). PMid:12184808. PMCid:PMC126239

Regan SL, McFarlane JR, O’Shea T, et al. Flow cytometric quantification of receptors and apoptosis in granulosa cells and ovulation rate. Reproduction (Cambridge, England). 2015. PMid:25948249

Campbell B, Marsters P, Baird D. The mechanism of action of the FecB (Booroola) mutation. Use of the FecB (Booroola) gene in sheep-breeding programs; 2009. p. 46

Jones ASK, Shikanov A. Follicle development as an orchestrated signaling network in a 3D organoid. Journal of Biological Engineering. 2019; 13(1):2. PMid:30647770. PMCid:PMC6327556

Murphy B. Equine chorionic gonadotropin: an enigmatic but essential tool. Animal Reproduction. 2012; 9(3):223–30.

Findlay JK, Drummond AE. Regulation of the FSH Receptor in the Ovary. Trends in endocrinology and metabolism: TEM. 1999; 10(5):183–8.

Zhang Z, Zhu Y, Lai Y, et al. Follicle-stimulating hormone inhibits apoptosis in ovarian cancer cells by regulating the OCT4 stem cell signaling pathway. International Journal of Oncology. 2013; 43(4):1194–204. PMid:23921511. PMCid:PMC3981007

Shimasaki S, Moore RK, Otsuka F, Erickson GF. The bone morphogenetic protein system in mammalian reproduction. Endocrine Reviews. 2004; 25(1):72–101. PMid:14769828

Khalaf M, Morera J, Bourret A, et al. BMP system expression in GCs from polycystic ovary syndrome women and the in vitro effects of BMP4, BMP6, and BMP7 on GC steroidogenesis. European Journal of Endocrinology/European Federation of Endocrine Societies. 2013; 168(3):437–44. PMid:23243014

Otsuka F, McTavish KJ, Shimasaki S. Integral role of GDF-9 and BMP-15 in ovarian function. Molecular Reproduction and Development. 2011; 78(1):9–21. PMid:21226076. PMCid:PMC3051839

Yi SE, LaPolt PS, Yoon BS, et al. The type I BMP receptor BmprIB is essential for female reproductive function. Proceedings of the National Academy of Sciences of the United States of America. 2001; 98(14):7994–9. PMid:11416163. PMCid:PMC35456

Regan SL, Knight PG, Yovich J, et al. Dysregulation of granulosal bone morphogenetic protein receptor 1B density is associated with reduced ovarian reserve and the age-related decline in human fertility. Molecular and Cellular Endocrinology. 2016. PMid:26805635