Expression of inhibin-activin subunits, follistatin and smads in granulosa-luteal cells collected at oocyte retrieval

  • Shiuh Young Chang
  • Hong-Yo Kang
  • Kuo-Chung Lan
  • Chang-Yi Hseh
  • Fu-Jen Huang
  • Ko-En Huang
Original Paper


Purpose: To examine the expression of inhibin-activin subunits, follistatin, and Smads 2, 3 and 4 in granulosa-luteal cells at the time of oocyte retrieval.

Methods: Quantitative reverse transcriptase-polymerase chain reaction was performed to quantify the mRNA expression of the investigated genes: inhibin α-subunit, inhibin-activin βA- and βB-subunits, follistatin, and Smads 2, 3 and 4.

Results: α-, βA- and βB-subunits of inhibin-activin, and follistatin were all well expressed in granulosa-luteal cells obtained from either dominant or cohort follicles. α-subunits strongly correlated with Smad2 (p < 0.001). βA-subunit significantly correlated with Smad4, p < 0.001. βB-subunit significantly correlated with Smad2, p=0.002, and follistatin expression strongly correlated with the expression of Smads 2 and 4 (p < 0.001 and = 0.007 respectively).

Conclusions: We found, for the first time that inhibin-activin βB-subunit mRNA was well expressed in human granulosa-luteal cells obtained from either dominant or cohort follicles. α-, βA- and βB-subunits and follistatin correlated with Smads to varied degrees, suggesting the active roles of the above genes at the time of oocyte retrieval.


Activin Follistatin Granulosa cells Inhibin Smad 



This study was supported by grant NSC 92-2314-B-182-084 from the National Science Council, Taiwan.


  1. 1.
    Findlay JK: An update on the roles of inhibin, activin, and follistatin as local regulators of folliculogenesis. Biol Eprod 1993;48:15–23.CrossRefGoogle Scholar
  2. 2.
    Knight PG, Glister C: Potential local regulatory functions of inhibins, activins and follistatin in the ovary. Reproduction 2001;121:503–12.CrossRefPubMedGoogle Scholar
  3. 3.
    Alak BM, Smith GD, Woodruff TK, Stouffer RL, Wolf DP: Enhancement of primate oocyte maturation and fertilization in vitro by inhibin A and activin A. Fertil Steril 1996;66:646–53.PubMedGoogle Scholar
  4. 4.
    Findlay JK, Drummond AE, Dyson ML, Baillie AJ, Robertson DM, Ethier JF: Recruitment and development of the follicle; the roles of the transforming growth factor-βsuperfamily. Mol Cell Endocrinol 2002;191:35–43.CrossRefPubMedGoogle Scholar
  5. 5.
    Heldin C-H, Miyazono K, ten Dijke P: TGF-β signaling from cell membrane to nucleus through Smad proteins. Nature 1997;390:465–71.CrossRefPubMedGoogle Scholar
  6. 6.
    Chapman SC, Bernard DJ, Jelen J, Woodruff TK: Properties of inhibin binding to betaglycan, InhBP/p20 and the activin type II receptors. Mol Cell Endocrinol 2002;196:79–93.CrossRefPubMedGoogle Scholar
  7. 7.
    Nakao A, Imamura T, Souchelnytskyi S, Kawabata M, Ishisaki A, Oeda E, et al.: TGF-β receptor-mediated signaling through Smad2, Smad3 and Smad4. EMBO J 1997;16:5353–62.CrossRefPubMedGoogle Scholar
  8. 8.
    Mukasa C, Nomura M, Tanaka T, Tanaka K, Nishi Y, Okabe T, et al.: Activin signaling through type IB activin receptor stimulates aromatase activity in the ovarian granulosa cell-like human granulosa (KGN) cells. Endocrinolgy 2003;144:1603–11.CrossRefGoogle Scholar
  9. 9.
    Marrs RP, Lobo R, Campeau JD, Nakamura RM, Brown J, Ujita EL, et al.: Correlation of human follicular fluid inhibin activity with spontaneous and induced follicle maturation. J Clin Endocrinol Metab 1984;58:704–9.PubMedGoogle Scholar
  10. 10.
    Chang SY, Lan K-C, Chen C-W, Huang F-J, Tsai M-Y, Chang C-Y: The influences of weather on patients with different ovarian responses in the treatment of assisted reproductive technology. J Assist Reprod Genetics 2005;22:191–8.CrossRefGoogle Scholar
  11. 11.
    Chang SY, Kang HY, Lan KC, Chang CY, Huang FJ, Tsai MY, et al.: Expression of steroid receptors, their cofactors, and aromatase in human luteinized granulosa cells after controlled ovarian hyperstimulation. Fertil Steril 2005;83 Suppl 1:1241–7.CrossRefPubMedGoogle Scholar
  12. 12.
    Schmitt C, Humeny A, Becker C-M, Brune K, Pahl A: Polymorphisms of TLR4: Rapid genotyping and reduced response to lipopolysaccharide of TLR4 mutant alleles. Clinical Chemistry 2002;48:1661–7.PubMedGoogle Scholar
  13. 13.
    Schwall RH, Mason AJ, Wilcox JN, Bassett SG, Zeleznik AJ: Localization of inhibin/activin subunit mRNAs within the primate ovary. Mol Endocrinol 1990;4:75–9.PubMedCrossRefGoogle Scholar
  14. 14.
    Welt CK, Smith ZA, Pauler DK, Hall JE: Differential regulation of inhibin A and inhibin B by luteinizing hormone, follicle-stimulating hormone, and stage of follicle development. J Clin Endocrinol Metab 2001;86:2531–7.Google Scholar
  15. 15.
    Roberts VJ, Barth S, El-Roeiy A, Yen SS: Expression of inhibin/activin subunits and follistatin messenger ribonucleic acids and proteins in ovarian follicles and the corpus luteum during the human menstrual cycle. J Clin Endocrinol Metab 1993;77:1402–10.CrossRefPubMedGoogle Scholar
  16. 16.
    Erämaa M, Heikinheimo K, Tuuri T, Hildén K, Ritvos O: Inhibin/activin subunit mRNA expression in human granulosa-luteal cells. Mol Cell Endocrinology 1993;92:R15–20.CrossRefGoogle Scholar
  17. 17.
    Magoffin DA, Jakimiuk AJ: Inhibin A, inhibin B and activin A in the follicular fluid of regularly cycling women. Hum Reprod 1997;12:1714–9.CrossRefPubMedGoogle Scholar
  18. 18.
    Davis AJ, Brooks CF, Johnson PA: Activin A and gonadotropin regulation of follicle-stimulating hormone and luteinizing hormone receptor messenger RNA in avian granulosa cells. Biol Reprod 2001;65:1352–8.CrossRefPubMedGoogle Scholar
  19. 19.
    Roh JS, Bondestam J, Mazerbourg S, Kaivo-Oja N, Groome N, Ritvos O, et al.: Endocrinology 2003;144:172–8.CrossRefPubMedGoogle Scholar
  20. 20.
    Erämaa M, Hildén K, Tuuri T, Ritvos O: Regulation of inhibin/activin subunit messenger ribinucleic acids (mRNAs) by activin A and expression of activin receptor mRNAs in cultured human granulosa-luteal cells. Endocrinology 1995;136:4382–9.CrossRefPubMedGoogle Scholar
  21. 21.
    Liu J, Hydèn-Granskog C, Voutilainen R: Gonadotrophins inhibit and activin induces expression of inhibin/activin βB subunit mRNA in cultured human granulose-luteal cells. Mol Hum Reprod 2001;7:319–23.CrossRefPubMedGoogle Scholar
  22. 22.
    Schipper I, Fauser BCJM, van Gaver EBO, Zarutskie PW, Dahl KD: Development of a human granulosa cell culture model with follicle-stimulating hormone responsiveness. Hum Reprod 1993;8:1380–6.PubMedGoogle Scholar
  23. 23.
    Wittmaack FM, Kreger DO, Blasco L, Tureck RW, Mastroianni L, Lessey BA: Effect of follicular size on oocyte retrieval, fertilization, cleavage, and embryo quality in in vitro fertilization cycles: a 6-year data collection. Fertil Steril 1994;62:1205–10.PubMedGoogle Scholar
  24. 24.
    Welt CK, Schneyer AL: Differential regulation of inhibin B and inhibin A by follicle-stimulating hormone and local growth factors in human granulosa cells from small antral follicles. J Clin Endocrinol Metabo 2001;86:330–6.CrossRefGoogle Scholar
  25. 25.
    McLachlan RI, Cohen NL, Vale WW, Rivier JE, Burger HG, Bremner WJ, et al.: The importance of luteinizing hormone in the control of inhibin and progesterone secretion by the human corpus luteum. J Clin Endocrinol Metabo 1989;68:1078–85.Google Scholar
  26. 26.
    Muttukrishna S, Groome N, Ledger W: Gonadotropic control of secretion of dimeric inhibins and activin A by human granulosa-luteal cells in vitro. J Assisted Reprod Genetics 1997;14:566–74.CrossRefGoogle Scholar
  27. 27.
    Schneyer AL, Fujiwara T, Fox J, Welt CK, Adams J, Messerlian GM, et al.: Dynamic changes in the intrafollicular inhibin/activin/follistatin axis during human follicular development: Relationship to circulating hormone concentrations. J Clin Endocrinol Metab 2000;85:3319–30.CrossRefPubMedGoogle Scholar
  28. 28.
    Sugawara M, DePaolo L, Nakatani A, DiMarzo S, Ling N: Radioimmunoassay of follistatin: application for in vitro fertilization procedures. J Clin Endocrinol Metab 1990;71:1672–4.PubMedCrossRefGoogle Scholar
  29. 29.
    Michel U, McMaster JW, Findlay JK: Regulation of steady-state mRNA levels in rat granulosa cells in vitro. J Mol Endocrinol 1992;9:147–56.PubMedGoogle Scholar
  30. 30.
    Li W, Khorasheh S, Yuen BH, Ling N, Leung PC: Stimulation of progesterone secretion by recombinant follistatin-288 in human granulosa cells. Endocrinology 1993;132:1750–6.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science&#x002B;Business Media, LLC 2006

Authors and Affiliations

  • Shiuh Young Chang
    • 1
    • 2
  • Hong-Yo Kang
    • 2
    • 3
  • Kuo-Chung Lan
    • 1
    • 2
  • Chang-Yi Hseh
    • 3
  • Fu-Jen Huang
    • 1
    • 2
  • Ko-En Huang
    • 1
    • 2
    • 3
  1. 1.Department of Obstetrics and GynecologyChang Gung Memorial Hospital-Kaohsiung Medical CenterKaohsiung CountyTaiwan
  2. 2.Chang Gung University College of MedicineTaoyuanTaiwan
  3. 3.Center of Reproductive MedicineChang Gung Memorial HospitalKaohsiungTaiwan

Personalised recommendations