Ligands, Receptors, and Transcription Factors that Mediate Inter-Cellular and Intra-Cellular Communication during Ovarian Follicle Development


Reliably producing a competent oocyte entails a deeper comprehension of ovarian follicle maturation, a very complex process that includes meiotic maturation of the female gamete, the oocyte, together with the mitotic divisions of the hormone-producing somatic cells. In this report, we investigate murine ovarian folliculogenesis in vivo using publicly available time-series microarrays from primordial to antral stage follicles. Manually curated protein interaction networks were employed to identify autocrine and paracrine signaling between the oocyte and the somatic cells (granulosa and theca cells) at multiple stages of follicle development. We established plausible protein-binding interactions between expressed genes that encode secreted factors and expressed genes that encode cellular receptors. Some computationally identified signaling interactions are well established, such as the paracrine signaling from the oocyte to the somatic cells through the oocyte-secreted growth factor Gdf9, while others are novel connections in term of ovarian folliculogenesis, such as the possible paracrine connection from somatic-secreted factor Ntn3 to the oocyte receptor Neo1. Additionally, we identified several of the likely transcription factors that might control the dynamic transcriptome during ovarian follicle development, noting that the YAP/TAZ signaling pathway is very active in vivo. This novel dynamic model of signaling and regulation can be employed to generate testable hypotheses regarding follicle development that could be validated experimentally, guiding the improvement of culture media to enhance in vitro ovarian follicle maturation and possibly novel therapeutic targets for reproductive diseases.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 510

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. 1.

    O'Brien MJ, Pendola JK, Eppig JJ. A revised protocol for in vitro development of mouse oocytes from primordial follicles dramatically improves their developmental competence. Biol Reprod. 2003;68(5):1682–6.

  2. 2.

    Eppig JJ, OBrien MJ. Development in vitro of mouse oocytes from primordial follicles. Biol Reprod. 1996;54(1):197–207.

  3. 3.

    Xu M, West E, Shea LD, Woodruff TK. Identification of a stage-specific permissive in vitro culture environment for follicle growth and oocyte development. Biol Reprod. 2006;75(6):916–23.

  4. 4.

    Edson MA, Nagaraja AK, Matzuk MM. The mammalian ovary from genesis to revelation. Endocrine reviews. 2009;30(6):624–712.

  5. 5.

    Xiao S, Coppeta JR, Rogers HB, et al. A microfluidic culture model of the human reproductive tract and 28-day menstrual cycle. Nature Communications. 2017;8.

  6. 6.

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

  7. 7.

    Uhlenhaut NH, Treier M. Foxl2 - function in ovarian development. Mol Genet Metab. 2006;88(3):225–34.

  8. 8.

    Chronowska E. High-Throughput Analysis of Ovarian Granulosa Cell Transcriptome. Biomed Res Int. 2014.

  9. 9.

    Yoon SJ, Kim KH, Chung HM, et al. Gene expression profiling of early follicular development in primordial, primary, and secondary follicles. Fertil Steril. 2006;85(1):193–203.

  10. 10.

    Skory RM, Bernabe BP, Galdones E, Broadbelt LJ, Shea LD, Woodruff TK. Microarray analysis identifies COMP as the most differentially regulated transcript throughout in vitro follicle growth. Mol Reprod Dev. 2013;80(2):132–44.

  11. 11.

    Wigglesworth K, Lee KB, Emori C, Sugiura K, Eppig JJ. Transcriptomic diversification of developing cumulus and mural granulosa cells in mouse ovarian follicles. Biol Reprod. 2015;92(1):23.

  12. 12.

    Pan H, O'Brien MJ, Wigglesworth K, Eppig JJ, Schultz RM. Transcript profiling during mouse oocyte development and the effect of gonadotropin priming and development in vitro. Dev Biol. 2005;286(2):493–506.

  13. 13.

    Grant CE, Bailey TL, Noble WS. FIMO: scanning for occurrences of a given motif. Bioinformatics. 2011;27(7):1017–8.

  14. 14.

    Zhao Y, Stormo GD. Quantitative analysis demonstrates most transcription factors require only simple models of specificity. Nat Biotechnol. 2011;29(6):480–3.

  15. 15.

    Xenarios I, Rice DW, Salwinski L, Baron MK, Marcotte EM, Eisenberg D. DIP: the database of interacting proteins. Nucleic acids research. 2000;28(1):289–91.

  16. 16.

    Penalver Bernabe B, Thiele I, Galdones E, et al. Dynamic genome-scale cell-specific metabolic models reveal novel inter-cellular and intra-cellular metabolic communications during ovarian follicle development. BMC Bioinformatics. 2019;20(1):307.

  17. 17.

    Shannon P, Markiel A, Ozier O, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498–504.

  18. 18.

    Xie XH, Lu J, Kulbokas EJ, et al. Systematic discovery of regulatory motifs in human promoters and 3 ' UTRs by comparison of several mammals. Nature. 2005;434(7031):338–45.

  19. 19.

    Wingender E, Dietze P, Karas H, Knuppel R. TRANSFAC: a database on transcription factors and their DNA binding sites. Nucleic Acids Res. 1996;24(1):238–41.

  20. 20.

    Matys V, Fricke E, Geffers R, et al. TRANSFAC: transcriptional regulation, from patterns to profiles. Nucleic Acids Res. 2003;31(1):374–8.

  21. 21.

    Weirauch MT, Yang A, Albu M, et al. Determination and inference of eukaryotic transcription factor sequence specificity. Cell. 2014;158(6):1431–43.

  22. 22.

    Leo CP, Vitt UA, Hsueh AJ. The Ovarian Kaleidoscope database: an online resource for the ovarian research community. Endocrinology. 2000;141(9):3052–4.

  23. 23.

    Schulz MH, Devanny WE, Gitter A, Zhong S, Ernst J, Bar-Joseph Z. DREM 2.0: Improved reconstruction of dynamic regulatory networks from time-series expression data. Bmc Syst Biol. 2012;6.

  24. 24.

    Sui SJH, Mortimer JR, Arenillas DJ, et al. oPOSSUM: identification of over-represented transcription factor binding sites in co-expressed genes. Nucleic Acids Res. 2005;33(10):3154–64.

  25. 25.

    Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Royal Stat Soc B. 1995:289–300.

  26. 26.

    Charlier C, Montfort J, Chabrol O, et al. Oocyte-somatic cells interactions, lessons from evolution. Bmc Genomics. 2012;13:560.

  27. 27.

    Adam M, Saller S, Strobl S, et al. Decorin is a part of the ovarian extracellular matrix in primates and may act as a signaling molecule. Hum Reprod. 2012;27(11):3249–58.

  28. 28.

    Kang JS, Yi MJ, Zhang W, Feinleib JL, Cole F, Krauss RS. Netrins and neogenin promote myotube formation. J Cell Biol. 2004;167(3):493–504.

  29. 29.

    Wang H, Copeland NG, Gilbert DJ, Jenkins NA, Tessier-Lavigne M. Netrin-3, a mouse homolog of human NTN2L, is highly expressed in sensory ganglia and shows differential binding to netrin receptors. J Neurosci. 1999;19(12):4938–47.

  30. 30.

    Lu X, Le Noble F, Yuan L, et al. The netrin receptor UNC5B mediates guidance events controlling morphogenesis of the vascular system. Nature. 2004;432(7014):179–86.

  31. 31.

    Larrivee B, Freitas C, Trombe M, et al. Activation of the UNC5B receptor by Netrin-1 inhibits sprouting angiogenesis. Genes Dev. 2007;21(19):2433–47.

  32. 32.

    Ozmadenci D, Feraud O, Markossian S, et al. Netrin-1 regulates somatic cell reprogramming and pluripotency maintenance. Nat Commun. 2015;6:7398.

  33. 33.

    Wilson NH, Key B. Neogenin: one receptor, many functions. Int J Biochem Cell Biol. 2007;39(5):874–8.

  34. 34.

    Buensuceso AV, Son AI, Zhou R, Paquet M, Withers BM, Deroo BJ. Ephrin-A5 Is required for optimal fertility and a complete ovulatory response to gonadotropins in the female mouse. Endocrinology. 2016;157(2):942–55.

  35. 35.

    Prunskaite-Hyyrylainen R, Shan J, Railo A, et al. Wnt4, a pleiotropic signal for controlling cell polarity, basement membrane integrity, and antimullerian hormone expression during oocyte maturation in the female follicle. FASEB J. 2014;28(4):1568–81.

  36. 36.

    Yang HS, Li Y, Deng HX, Peng F. Identification of beta2-microglobulin as a potential target for ovarian cancer. Cancer Biol Ther. 2009;8(24):2323–8.

  37. 37.

    Qian J, Ji F, Ye X, et al. IGHG1 promotes motility likely through epithelial-mesenchymal transition in ovarian cancer. Chin J Cancer Res. 2018;30(2):282–90.

  38. 38.

    Chang CL, Wang HS, Soong YK, Huang SY, Pai SY, Hsu SY. Regulation of oocyte and cumulus cell interactions by intermedin/adrenomedullin 2. J Biol Chem. 2011;286(50):43193–203.

  39. 39.

    Dickinson RE, Hryhorskyj L, Tremewan H, et al. Involvement of the SLIT/ROBO pathway in follicle development in the fetal ovary. Reproduction. 2010;139(2):395–407.

  40. 40.

    Dissen GA, Hirshfield AN, Malamed S, Ojeda SR. Expression of neurotrophins and their receptors in the mammalian ovary is developmentally-regulated - changes at the time of folliculogenesis. Endocrinology. 1995;136(10):4681–92.

  41. 41.

    Dissen GA, Romero C, Hirshfield AN, Ojeda SR. Nerve growth factor is required for early follicular development in the mammalian ovary. Endocrinology. 2001;142(5):2078–86.

  42. 42.

    Pedersen T, Peters H. Proposal for a classification of oocytes and follicles in the mouse ovary. J Reprod Fertil. 1968;17(3):555–7.

  43. 43.

    Bridges PJ, Cho J, Ko C. Endothelins in regulating ovarian and oviductal function. Front Biosci (Schol Ed). 2011;3:145–55.

  44. 44.

    McGee EA, Hsueh AJ. Initial and cyclic recruitment of ovarian follicles. Endocr Rev. 2000;21(2):200–14.

  45. 45.

    Wood JR, Strauss JF 3rd. Multiple signal transduction pathways regulate ovarian steroidogenesis. Rev Endocr Metab Disord. 2002;3(1):33–46.

  46. 46.

    Jensen CE, Zachariae F. Studies on the mechanism of ovulation: isolation and analysis of acid mucopolysaccharides in bovine follicular fluid. Acta Endocrinol. 1958;27(3):356–68.

  47. 47.

    Gebauer H, Lindner HR, Amsterdam A. Synthesis of heparin-like glycosaminoglycans in rat ovarian slices. Biol Reprod. 1978;18(3):350–8.

  48. 48.

    Eriksen GV, Carlstedt I, Morgelin M, Uldbjerg N, Malmstrom A. Isolation and characterization of proteoglycans from human follicular fluid. Biochem J. 1999;340:613–20.

  49. 49.

    Young JM, McNeilly AS. Theca: the forgotten cell of the ovarian follicle. Reproduction. 2010;140(4):489–504.

  50. 50.

    Mehlmann LM. Stops and starts in mammalian oocytes: recent advances in understanding the regulation of meiotic arrest and oocyte maturation. Reproduction. 2005;130(6):791–9.

  51. 51.

    Dickinson RE, Myers M, Duncan WC. Novel regulated expression of the SLIT/ROBO pathway in the ovary: possible role during luteolysis in women. Endocrinology. 2008;149(10):5024–34.

  52. 52.

    Pyun JA, Cha DH, Kwack K. LAMC1 gene is associated with premature ovarian failure. Maturitas. 2012;71(4):402–6.

  53. 53.

    Kaneko T, Saito H, Toya M, Satio T, Nakahara K, Hiroi M. Hyaluronic acid inhibits apoptosis in granulosa cells via CD44. J Assist Reprod Gen. 2000;17(3):162–7.

  54. 54.

    Albertini DF, Sanfins A, Combelles CM. Origins and manifestations of oocyte maturation competencies. Reprod BioMed Online. 2003;6(4):410–5.

  55. 55.

    Zuccotti M, Piccinelli A, Rossi PG, Garagna S, Redi CA. Chromatin organization during mouse oocyte growth. Mol Reprod Dev. 1995;41(4):479–85.

  56. 56.

    De la Fuente R, Eppig JJ. Transcriptional activity of the mouse oocyte genome: companion granulosa cells modulate transcription and chromatin remodeling. Dev Biol. 2001;229(1):224–36.

  57. 57.

    Jang YJ, Park JI, Moon WJ, Dam PTM, Cho MK, Chun SY. Cumulus cell-expressed type I interferons induce cumulus expansion in mice. Biol Reprod. 2015;92(1).

  58. 58.

    Sirotkin AV. Transcription factors and ovarian functions. J Cell Physiol. 2010;225(1):20–6.

  59. 59.

    Du H, Taylor HS. The role of hox genes in female reproductive tract development, adult function, and fertility. Cold Spring Harb Perspect Med. 2015;6(1):a023002.

  60. 60.

    Lakhwani S, Garcia-Sanz P, Vallejo M. Alx3-deficient mice exhibit folic acid-resistant craniofacial midline and neural tube closure defects. Dev Biol. 2010;344(2):869–80.

  61. 61.

    Zhang CP, Yang JL, Zhang J, et al. Notch signaling is involved in ovarian follicle development by regulating granulosa cell proliferation. Endocrinology. 2011;152(6):2437–47.

  62. 62.

    Pisarska MD, Barlow G, Kuo FT. Minireview: roles of the forkhead transcription factor FOXL2 in granulosa cell biology and pathology. Endocrinology. 2011;152(4):1199–208.

  63. 63.

    Chu S, Nishi Y, Yanase T, Nawata H, Fuller PJ. Transrepression of estrogen receptor beta signaling by nuclear factor-kappab in ovarian granulosa cells. Mol Endocrinol. 2004;18(8):1919–28.

  64. 64.

    Worrad DM, Ram PT, Schultz RM. Regulation of gene expression in the mouse oocyte and early preimplantation embryo: developmental changes in Sp1 and TATA box-binding protein. TBP Development. 1994;120(8):2347–57.

  65. 65.

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

  66. 66.

    Woodruff TK, Shea LD. The role of the extracellular matrix in ovarian follicle development. Reprod Sci. 2007;14(8 Suppl):6–10.

  67. 67.

    Gallardo TD, John GB, Shirley L, et al. Genomewide discovery and classification of candidate ovarian fertility genes in the mouse. Genetics. 2007;177(1):179–94.

  68. 68.

    Buensuceso AV, Deroo BJ. The ephrin signaling pathway regulates morphology and adhesion of mouse granulosa cells in vitro. Biology of reproduction. 2013;88(1):25.

  69. 69.

    Hatzirodos N, Hummitzsch K, Irving-Rodgers HF, Harland ML, Morris SE, Rodgers RJ. Transcriptome profiling of granulosa cells from bovine ovarian follicles during atresia. Bmc Genomics. 2014;15(40).

  70. 70.

    Hornick JE, Duncan FE, Shea LD, Woodruff TK. Multiple follicle culture supports primary follicle growth through paracrine-acting signals. Reproduction. 2013;145(1):19–32.

  71. 71.

    Lei QY, Zhang H, Zhao B, et al. TAZ promotes cell proliferation and epithelial-mesenchymal transition and is inhibited by the hippo pathway. Mol Cell Biol. 2008;28(7):2426–36.

  72. 72.

    Kawamura K, Cheng Y, Suzuki N, et al. Hippo signaling disruption and Akt stimulation of ovarian follicles for infertility treatment. Proc Natl Acad Sci U S A. 2013;110(43):17474–9.

  73. 73.

    Li J, Kawamura K, Cheng Y, et al. Activation of dormant ovarian follicles to generate mature eggs. Proc Natl Acad Sci U S A. 2010;107(22):10280–4.

  74. 74.

    Yu FX, Zhang YF, Park HW, et al. Protein kinase A activates the Hippo pathway to modulate cell proliferation and differentiation. Genes & development. 2013;27(11):1223–32.

  75. 75.

    Aplin AE, Howe AK, Juliano RL. Cell adhesion molecules, signal transduction and cell growth. Curr Opin Cell Biol. 1999;11(6):737–44.

  76. 76.

    Gerard C, Goldbeter A. The balance between cell cycle arrest and cell proliferation: control by the extracellular matrix and by contact inhibition. Interface Focus. 2014;4(3).

  77. 77.

    Weiss MS, Penalver Bernabe B, Shin S, et al. Dynamic transcription factor activity and networks during ErbB2 breast oncogenesis and targeted therapy. Integr Biol (Camb). 2014;6(12):1170–82.

  78. 78.

    Zhou H, Decker JT, Lemke MM, et al. Synergy of paracrine signaling during early-stage mouse ovarian follicle development in vitro. Cell Mol Bioeng. 2018;11(5):435–50.

Download references


We thank Dr. Lei Lei, Sarah Kiesewetter for their invaluable technical support and Dr. Nadereh Jafari, Director of the Genomics Core Facility (Center for Genetic Medicine, Northwestern University) and Dr. Simon Lin and Dr. Gang Feng. Additionally, we would like to thank Dr. Ariella Shikanov and Andrea Jones for their insightful comments and throughout editing this manuscript.


This work has been mainly supported by NIH/NICHD 2 U54 HD041857–07 for study design and collection, analysis and interpretation of the data. BPB was supported NIH/NIGMS 2 T32 GM008449–16.

Author information

BPB designed the computational approached and performed all the computational study. BPB, TKW, LJB and LDS interpreted the results and wrote the manuscript.

Correspondence to Lonnie D. Shea.

Ethics declarations

Conflict of Interests

The authors declare that they have no conflict of interest.

Ethics Approval

Animals were treated in accordance with the NIH Guide for the Care and Use of Laboratory Animals and the established IACUC protocol at Northwestern University.

Consent to Publish

Not applicable.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material


(DOCX 749 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Bernabé, B.P., Woodruff, T., Broadbelt, L.J. et al. Ligands, Receptors, and Transcription Factors that Mediate Inter-Cellular and Intra-Cellular Communication during Ovarian Follicle Development. Reprod. Sci. (2020) doi:10.1007/s43032-019-00075-8

Download citation


  • Dynamic signaling
  • Dynamic regulation ovarian follicle development
  • Inter-cellular communication