Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Involvement of activin signaling in abnormalities of mouse vagina exposed neonatally to diethylstilbestrol

  • 172 Accesses

  • 5 Citations


Perinatal exposure to a synthetic estrogen, diethylstilbestrol (DES), causes cervicovaginal adenosis and permanent hyperplastic cornified vaginal epithelium with keratinization in mice. To investigate the mechanisms of the induction of vaginal abnormalities by DES, we have focused on activin A signaling. We have found that the βA-subunit mRNA is mainly expressed in the neonatal vaginal stroma, whereas activin A receptor type IB is localized in the neonatal vaginal epithelium. SMAD2, the intracellular signaling protein, is phosphorylated in the neonatal vagina. Cell proliferation in the vaginal epithelium grown in vitro is reduced by DES treatment or by activin signaling suppression through inhibin treatment. Thus, activin A (a homodimer of the βA-subunit) in the stroma stimulates epithelial cell proliferation in the neonatal vagina. DES treatment decreases the expression of the βA-subunit and activin receptor IIB but increases the expression of the βB-subunit and inhibin receptor. Neonatal DES treatment inhibits the phosphorylation of SMAD2 in the vaginal epithelium, indicating the inhibition of activin A signaling in the vaginal epithelium by neonatal DES treatment. Treatment with DES or inhibin, a native antagonist of activin, induces adenosis-like structures and keratinization in the vagina grown in vitro. These data suggest that the suppression of activin A signaling by DES is involved in the induction of cervicovaginal adenosis and keratinization in the neonatal mouse vaginal epithelium.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7


  1. Ball EM, Risbridger GP (2001) Activins as regulators of branching morphogenesis. Dev Biol 238:1–12

  2. Bigsby RM, Cooke PS, Cunha GR (1986) A simple efficient method for separating murine uterine epithelial and mesenchymal cells. Am J Physiol 251: E630-E636

  3. Brown CW, Houston-Hawkins DE, Woodruff TK, Matzuk MM (2000) Insertion of Inhbb into the Inhba locus rescues the Inhba-null phenotype and reveals new activin functions. Nat Genet 25:453–457

  4. Chabicovsky M, Herkner K, Rossmanith W (2003) Overexpression of activin β(C) or activin β(E) in the mouse liver inhibits regenerative deoxyribonucleic acid synthesis of hepatic cells. Endocrinology 144:3497–3504

  5. Chang H, Lau AL, Matzuk MM (2001) Studying TGF-β superfamily signaling by knockouts and knockins. Mol Cell Endocrinol 180:39–46

  6. Chang H, Brown CW, Matzuk MM (2002) Genetic analysis of the mammalian transforming growth factor-β superfamily. Endocr Rev 23:787–823

  7. Cook RW, Thompson TB, Jardetzky TS, Woodruff TK (2004) Molecular biology of inhibin action. Semin Reprod Med 22:269–276

  8. Couse JF, Korach KS (2004) Estrogen receptor-α mediates the detrimental effects of neonatal diethylstilbestrol (DES) exposure in the murine reproductive tract. Toxicology 205:55–63

  9. Forsberg JG (1979) Developmental mechanism of estrogen-induced irreversible changes in the mouse cervicovaginal epithelium. Natl Cancer Inst Monogr 51:41–56

  10. Giusti RM, Iwamoto K, Hatch EE (1995) Diethylstilbestrol revisited: a review of the long-term health effects. Ann Intern Med 122:778–788

  11. Herbst AL, Ulfelder H, Poskanzer DC (1971) Adenocarcinoma of the vagina. Association of maternal stilbestrol therapy with tumor appearance in young women. N Engl J Med 284:878–881

  12. Iguchi T (1992) Cellular effects of early exposure to sex hormones and abnormalities of female reproduction. Int Rev Cytol 139:1–57

  13. Iguchi T, Sato T (2000) Endocrine disruption and developmental abnormalities of female reproduction. Am Zool 40:402–411

  14. Iguchi T, Takasugi N (1987) Postnatal development of uterine abnormalities in mice exposed to DES in utero. Biol Neonate 52:97–103

  15. Iguchi T, Takase M, Takasugi N (1986) Development of vaginal adenosis-like lesions and uterine epithelial stratification in mice exposed perinatally to diethylstilbestrol. Proc Soc Exp Biol Med 181:59–65

  16. Jones RL, Stoikos C, Findlay JK, Salamonsen LA (2006) TGF-β superfamily expression and actions in the endometrium and placenta. Reproduction 132:217–232

  17. Kim H, Nakajima T, Hayashi S, Chambon P, Watanabe H, Iguchi T, Sato T (2009) Effects of diethylstilbestrol on programmed oocyte death and induction of polyovular follicles in neonatal mouse ovaries. Biol Reprod 81:1002–1009

  18. Kipp JL, Kilen SM, Bristol-Gould S, Woodruff TK, Mayo KE (2007) Neonatal exposure to estrogens suppresses activin expression and signaling in the mouse ovary. Endocrinology 148:1968–1976

  19. Knight PG, Glister C (2001) Potential local regulatory functions of inhibins, activins and follistatin in the ovary. Reproduction 121:503–512

  20. Kurita T, Mills AA, Cunha GR (2004) Roles of p63 in the diethylstilbestrol-induced cervicovaginal adenosis. Development 131:1639–1649

  21. Miyagawa S, Buchanan DL, Sato T, Ohta Y, Nishina Y, Iguchi T (2002) Characterization of diethylstilbestrol-induced hypospadias in female mice. Anat Rec 266:43–50

  22. Miyagawa S, Katsu Y, Watanabe H, Iguchi T (2004a) Estrogen-independent activation of erbBs signaling and estrogen receptor alpha in the mouse vagina exposed neonatally to diethylstilbestrol. Oncogene 23:340–349

  23. Miyagawa S, Suzuki A, Katsu Y, Kobayashi M, Goto M, Handa H, Watanabe H, Iguchi T (2004b) Persistent gene expression in mouse vagina exposed neonatally to diethylstilbestrol. J Mol Endocrinol 32:663–677

  24. Mori T, Iguchi T (1988) Long-term effects of perinatal treatment with sex steroids and related substances on reproductive organs of female mice. In: Mori T, Nagasawa H (eds) Toxicity of hormones in perinatal life. CRC Press, Boca Raton, pp 63–79

  25. Murata K, Ota S, Niki T, Goto A, Li CP, Ruriko UM, Ishikawa S, Aburatani H, Kuriyama T, Fukayama M (2007) p63—Key molecule in the early phase of epithelial abnormality in idiopathic pulmonary fibrosis. Exp Mol Pathol 83:367–376

  26. Niimi S, Hyuga M, Kazama H, Inagawa M, Seki T, Ariga T, Kobayashi T, Hayakawa T (2002a) Activins A, AB, and B inhibit hepatocyte growth factor synthesis by MRC-5 human lung fibroblasts. Biol Pharm Bull 25:1405–1408

  27. Niimi S, Horikawa M, Seki T, Ariga T, Kobayashi T, Hayakawa T (2002b) Effect of activins AB and B on DNA synthesis stimulated by epidermal growth factor in primary cultured rat hepatocytes. Biol Pharm Bull 25:437–440

  28. Ozawa S, Iguchi T, Sawada K, Ohta Y, Takasugi N, Bern HA (1991) Postnatal vaginal nodules induced by prenatal diethylstilbestrol treatment correlate with later development of ovary-independent vaginal and uterine changes in mice. Cancer Lett 58:167–175

  29. Pangas SA, Woodruff TK (2000) Activin signal transduction pathways. Trends Endocrinol Metab 11:309–314

  30. Perttu MC, Martikainen PM, Huhtala HS, Bläuer M, Tammela TL, Tuohimaa PJ, Syvälä H (2006) Altered levels of Smad2 and Smad4 are associated with human prostate carcinogenesis. Prostate Cancer Prostatic Dis 9:185–189

  31. Risbridger GP, Schmitt JF, Robertson DM (2001) Activins and inhibins in endocrine and other tumors. Endocr Rev 22:836–858

  32. Robboy SJ, Szyfelbein WM, Goellner JR, Kaufman RH, Taft PD, Richard RM, Gaffey TA, Prat J, Virata R, Hatab PA, McGorray SP, Noller KL, Townsend D, Labarthe D, Barnes AB (1981) Dysplasia and cytologic findings in 4,589 young women enrolled in diethylstilbestrol-adenosis (DESAD) project. Am J Obstet Gynecol 140:579–586

  33. Simon DP, Vadakkadath Meethal S, Wilson AC, Gallego MJ, Weinecke SL, Bruce E, Lyons PF, Haasl RJ, Bowen RL, Atwood CS (2009) Activin receptor signaling regulates prostatic epithelial cell adhesion and viability. Neoplasia 11:365–376

  34. Suzuki A, Sugihara A, Uchida K, Sato T, Ohta Y, Katsu Y, Watanabe H, Iguchi T (2002) Developmental effects of perinatal exposure to bisphenol-A and diethylstilbestrol on reproductive organs in female mice. Reprod Toxicol 16:107–116

  35. Suzuki A, Watanabe H, Mizutani T, Sato T, Ohta Y, Iguchi T (2006) Global gene expression in mouse vaginae exposed to diethylstilbestrol at different ages. Exp Biol Med 231:632–640

  36. Suzuki A, Urushitani H, Sato T, Kobayashi T, Watanabe H, Ohta Y, Iguchi T (2007) Gene expression change in the Müllerian duct of the mouse fetus exposed to diethylstilbestrol in utero. Exp Biol Med 232:503–514

  37. Thompson TB, Cook RW, Chapman SC, Jardetzky TS, Woodruff TK (2004) Beta A versus beta B: is it merely a matter of expression? Mol Cell Endocrinol 225:9–17

  38. Truong AB, Khavari PA (2007) Control of keratinocyte proliferation and differentiation by p63. Cell Cycle 6:295–299

  39. Waltermann A, Kartasheva NN, Dobbelstein M (2003) Differential regulation of p63 and p73 expression. Oncogene 22:5686–5693

  40. Yang J, Wahdan-Alaswad R, Danielpour D (2009) Critical role of Smad2 in tumor suppression and transforming growth factor-beta-induced apoptosis of prostate epithelial cells. Cancer Res 69:2185–2190

Download references


We thank Dr. Raphael Guzman, Department of Molecular and Cell Biology and the Cancer Research Laboratory, University of California, Berkeley, USA, for his critical reading of the manuscript.


The authors declare no conflicts of interest.

Author information

Correspondence to Tomomi Sato.

Additional information

This work was partially supported by a Grant-in-Aid for Scientific Research (B) (T.I.) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, Grants for Strategic Research Projects at Yokohama City University (nos. K19020, K2109, G2201, and W18005 to T.S.), a Health Sciences Research Grant from the Ministry of Health, Labor, and Welfare, Japan (T.I.), and a grant for Support of the Collaborative Study at NIBB (T.S).

Electronic supplementary material

Below is the link to the electronic supplementary material.

Fig. S

The expression of inhibin α, βA-subunit, follistatin (Fst), activin receptor (Acvr) 1, Acvr1b, Acvr2a, Acvr2b and transforming growth factor, β receptor III (Tgfβr3) mRNAs in the uterine epithelium (UE), uterine stroma (US), vaginal epithelium (VE) and vaginal stroma (VS) at days 2 and 15 was analyzed by RT-PCR. Peptidylprolyl isomerase A (Ppia) was chosen as an internal standard (GIF 62 kb)

High resolution (TIFF 6099 kb)

Table S

(DOC 99 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nakajima, T., Iguchi, T. & Sato, T. Involvement of activin signaling in abnormalities of mouse vagina exposed neonatally to diethylstilbestrol. Cell Tissue Res 344, 527 (2011). https://doi.org/10.1007/s00441-011-1161-2

Download citation


  • Diethylstilbestrol
  • Activin
  • Uterus
  • Vagina
  • Cervicovaginal adenosis
  • Mouse