Skip to main content

Advertisement

SpringerLink
Log in

Expression of inhibitor proteins that control primordial follicle reserve decreases in cryopreserved ovaries after autotransplantation

  • Fertility Preservation
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Even with 86 live births reported globally so far, the mechanism of primordial follicle loss following autotransplantation of the frozen-thawed ovarian tissue needs further evaluation. Pten, Tsc1, p27, and Amh are the inhibitor proteins that play crucial roles in suppressing the transition from the primordial follicle to primary state, maintaining the primordial follicle reserve. In this study, we aimed to evaluate whether the expression patterns of these proteins change and it may be related to the global primordial follicle loss after autotransplantation of the frozen-thawed ovarian tissue.

Methods

Four groups were established in rats: fresh-control, frozen/thawed, fresh-transplanted, and frozen/thawed and transplanted. After slow freezing and thawing process, two ovarian pieces were transplanted into the back muscle of the same rat. After 2 weeks, grafts were harvested, fixed, and embedded into the paraffin block. Normal and atretic primordial/growing follicle count was performed in all groups. Ovarian tissues were evaluated for the dynamic expressions of the Pten, Tsc1, p27, and Amh proteins using immunohistochemistry, and H-score analyses were done.

Results

Ovarian tissue cryopreservation does not change the expression patterns of inhibitory proteins that control ovarian reserve. Both in fresh and frozen/thawed autotransplanted groups, the expression of inhibitory proteins and Amh decreased significantly in primordial follicles and in growing follicles, respectively. In control group and in frozen/thawed group, primordial follicle counts were similar but decreased by almost half in both fresh-transplanted and frozen/thawed and transplanted groups.

Conclusions

One of the causes of primordial follicle loss after transplantation of ovarian graft may be decreased expression of the inhibitory proteins that guard the ovarian reserve and transplantation itself seems to be the major cause for disruption of inhibitory molecular signaling. Our findings highlight important molecular aspects for future clinical applications for fertility preservation in humans.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

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

Similar content being viewed by others

References

  1. Sonmezer M, Ozkavukcu S. Fertility preservation in females with malignant disease-1: causes, clinical needs and indications. Turk J Hematol. 2009;26.

  2. Lee SJ, Schover LR, Partridge AH, Patrizio P, Wallace WH, Hagerty K, et al. American Society of Clinical Oncology recommendations on fertility preservation in cancer patients. J Clin Oncol. 2006;24(18):2917–31. https://doi.org/10.1200/JCO.2006.06.5888.

    Article  PubMed  Google Scholar 

  3. Donnez J, Dolmans MM. Ovarian cortex transplantation: 60 reported live births brings the success and worldwide expansion of the technique towards routine clinical practice. J Assist Reprod Genet. 2015;32(8):1167–70. https://doi.org/10.1007/s10815-015-0544-9.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Fortune JE. The early stages of follicular development: activation of primordial follicles and growth of preantral follicles. Anim Reprod Sci. 2003;78(3–4):135–63.

    Article  CAS  PubMed  Google Scholar 

  5. Oktem O, Urman B. Understanding follicle growth in vivo. Hum Reprod. 2010;25(12):2944–54. https://doi.org/10.1093/humrep/deq275.

    Article  PubMed  Google Scholar 

  6. Adhikari D, Liu K. Molecular mechanisms underlying the activation of mammalian primordial follicles. Endocr Rev. 2009;30(5):438–64. https://doi.org/10.1210/er.2008-0048.

    Article  CAS  PubMed  Google Scholar 

  7. Carlsson IB, Scott JE, Visser JA, Ritvos O, Themmen AP, Hovatta O. Anti-Mullerian hormone inhibits initiation of growth of human primordial ovarian follicles in vitro. Hum Reprod. 2006;21(9):2223–7. https://doi.org/10.1093/humrep/del165.

    Article  CAS  PubMed  Google Scholar 

  8. Cantley LC. The phosphoinositide 3-kinase pathway. Science. 2002;296(5573):1655–7. https://doi.org/10.1126/science.296.5573.1655.

    Article  CAS  PubMed  Google Scholar 

  9. Suzuki A, de la Pompa JL, Stambolic V, Elia AJ, Sasaki T, del Barco BI, et al. High cancer susceptibility and embryonic lethality associated with mutation of the PTEN tumor suppressor gene in mice. Curr Biol. 1998;8(21):1169–78.

    Article  CAS  PubMed  Google Scholar 

  10. Reddy P, Liu L, Adhikari D, Jagarlamudi K, Rajareddy S, Shen Y, et al. Oocyte-specific deletion of Pten causes premature activation of the primordial follicle pool. Science. 2008;319(5863):611–3. https://doi.org/10.1126/science.1152257.

    Article  CAS  PubMed  Google Scholar 

  11. John GB, Gallardo TD, Shirley LJ, Castrillon DH. Foxo3 is a PI3K-dependent molecular switch controlling the initiation of oocyte growth. Dev Biol. 2008;321(1):197–204. https://doi.org/10.1016/j.ydbio.2008.06.017.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Goto M, Iwase A, Ando H, Kurotsuchi S, Harata T, Kikkawa F. PTEN and Akt expression during growth of human ovarian follicles. J Assist Reprod Genet. 2007;24(11):541–6. https://doi.org/10.1007/s10815-007-9156-3.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Shin I, Rotty J, Wu FY, Arteaga CL. Phosphorylation of p27Kip1 at Thr-157 interferes with its association with importin alpha during G1 and prevents nuclear re-entry. J Biol Chem. 2005;280(7):6055–63. https://doi.org/10.1074/jbc.M412367200.

    Article  CAS  PubMed  Google Scholar 

  14. Rajareddy S, Reddy P, Du C, Liu L, Jagarlamudi K, Tang W, et al. p27kip1 (cyclin-dependent kinase inhibitor 1B) controls ovarian development by suppressing follicle endowment and activation and promoting follicle atresia in mice. Mol Endocrinol. 2007;21(9):2189–202. https://doi.org/10.1210/me.2007-0172.

    Article  CAS  PubMed  Google Scholar 

  15. Kaldis P. Another piece of the p27Kip1 puzzle. Cell. 2007;128(2):241–4. https://doi.org/10.1016/j.cell.2007.01.006.

    Article  CAS  PubMed  Google Scholar 

  16. Crino PB, Nathanson KL, Henske EP. The tuberous sclerosis complex. N Engl J Med. 2006;355(13):1345–56. https://doi.org/10.1056/NEJMra055323.

    Article  CAS  PubMed  Google Scholar 

  17. Adhikari D, Zheng W, Shen Y, Gorre N, Hamalainen T, Cooney AJ, et al. Tsc/mTORC1 signaling in oocytes governs the quiescence and activation of primordial follicles. Hum Mol Genet. 2010;19(3):397–410. https://doi.org/10.1093/hmg/ddp483.

    Article  CAS  PubMed  Google Scholar 

  18. Baarends WM, Uilenbroek JT, Kramer P, Hoogerbrugge JW, van Leeuwen EC, Themmen AP, et al. Anti-mullerian hormone and anti-Mullerian hormone type II receptor messenger ribonucleic acid expression in rat ovaries during postnatal development, the estrous cycle, and gonadotropin-induced follicle growth. Endocrinology. 1995;136(11):4951–62. https://doi.org/10.1210/endo.136.11.7588229.

    Article  CAS  PubMed  Google Scholar 

  19. Durlinger AL, Kramer P, Karels B, de Jong FH, Uilenbroek JT, Grootegoed JA, et al. Control of primordial follicle recruitment by anti-Mullerian hormone in the mouse ovary. Endocrinology. 1999;140(12):5789–96. https://doi.org/10.1210/endo.140.12.7204.

    Article  CAS  PubMed  Google Scholar 

  20. Topal-Celikkan F, Ozkavukcu S, Balci D, Serin-Kilicoglu S, Atabenli-Erdemli E. Mouse ovarian tissue vitrification on copper electron microscope grids versus slow freezing: a comparative ultrastructural study. Reprod Fertil Dev. 2014; https://doi.org/10.1071/RD13262.

  21. Gandolfi F, Paffoni A, Papasso Brambilla E, Bonetti S, Brevini TA, Ragni G. Efficiency of equilibrium cooling and vitrification procedures for the cryopreservation of ovarian tissue: comparative analysis between human and animal models. Fertil Steril. 2006;85(Suppl 1):1150–6. https://doi.org/10.1016/j.fertnstert.2005.08.062.

    Article  PubMed  Google Scholar 

  22. Inan S, Vatansever S, Celik-Ozenci C, Sanci M, Dicle N, Demir R. Immunolocalizations of VEGF, its receptors flt-1, KDR and TGF-beta’s in epithelial ovarian tumors. Histol Histopathol. 2006;21(10):1055–64. https://doi.org/10.14670/HH-21.1055.

    CAS  PubMed  Google Scholar 

  23. Kawamura K, Kawamura N, Hsueh AJ. Activation of dormant follicles: a new treatment for premature ovarian failure? Curr Opin Obstet Gynecol. 2016;28(3):217–22. https://doi.org/10.1097/GCO.0000000000000268.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Silber SJ. Ovary cryopreservation and transplantation for fertility preservation. Mol Hum Reprod. 2012;18(2):59–67. https://doi.org/10.1093/molehr/gar082.

    Article  CAS  PubMed  Google Scholar 

  25. Nugent D, Newton H, Gallivan L, Gosden RG. Protective effect of vitamin E on ischaemia-reperfusion injury in ovarian grafts. J Reprod Fertil. 1998;114(2):341–6.

    Article  CAS  PubMed  Google Scholar 

  26. Israely T, Dafni H, Nevo N, Tsafriri A, Neeman M. Angiogenesis in ectopic ovarian xenotransplantation: multiparameter characterization of the neovasculature by dynamic contrast-enhanced MRI. Magn Reson Med. 2004;52(4):741–50. https://doi.org/10.1002/mrm.20203.

    Article  PubMed  Google Scholar 

  27. Van Eyck AS, Jordan BF, Gallez B, Heilier JF, Van Langendonckt A, Donnez J. Electron paramagnetic resonance as a tool to evaluate human ovarian tissue reoxygenation after xenografting. Fertil Steril. 2009;92(1):374–81. https://doi.org/10.1016/j.fertnstert.2008.05.012.

    Article  PubMed  Google Scholar 

  28. Demeestere I, Simon P, Emiliani S, Delbaere A, Englert Y. Orthotopic and heterotopic ovarian tissue transplantation. Hum Reprod Update. 2009;15(6):649–65. https://doi.org/10.1093/humupd/dmp021.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Van Eyck AS, Bouzin C, Feron O, Romeu L, Van Langendonckt A, Donnez J, et al. Both host and graft vessels contribute to revascularization of xenografted human ovarian tissue in a murine model. Fertil Steril. 2010;93(5):1676–85. https://doi.org/10.1016/j.fertnstert.2009.04.048.

    Article  PubMed  Google Scholar 

  30. Kim SS. Fertility preservation in female cancer patients: current developments and future directions. Fertil Steril. 2006;85(1):1–11. https://doi.org/10.1016/j.fertnstert.2005.04.071.

    Article  CAS  PubMed  Google Scholar 

  31. Li SH, Hwu YM, Lu CH, Chang HH, Hsieh CE, Lee RK. VEGF and FGF2 improve revascularization, survival, and oocyte quality of cryopreserved, subcutaneously-transplanted mouse ovarian tissues. Int J Mol Sci. 2016;17(8) https://doi.org/10.3390/ijms17081237.

  32. Gavish Z, Peer G, Roness H, Cohen Y, Meirow D. Follicle activation and ‘burn-out’ contribute to post-transplantation follicle loss in ovarian tissue grafts: the effect of graft thickness. Hum Reprod. 2014;29(5):989–96. https://doi.org/10.1093/humrep/deu015.

    Article  PubMed  Google Scholar 

  33. Gavish Z, Spector I, Peer G, Schlatt S, Wistuba J, Roness H, et al. Follicle activation is a significant and immediate cause of follicle loss after ovarian tissue transplantation. J Assist Reprod Genet. 2017; https://doi.org/10.1007/s10815-017-1079-z.

  34. Lopez-Neblina F, Toledo-Pereyra LH. Phosphoregulation of signal transduction pathways in ischemia and reperfusion. J Surg Res. 2006;134(2):292–9. https://doi.org/10.1016/j.jss.2006.01.007.

    Article  PubMed  Google Scholar 

  35. David A, Van Langendonckt A, Gilliaux S, Dolmans MM, Donnez J, Amorim CA. Effect of cryopreservation and transplantation on the expression of kit ligand and anti-Mullerian hormone in human ovarian tissue. Hum Reprod. 2012;27(4):1088–95. https://doi.org/10.1093/humrep/des013.

    Article  CAS  PubMed  Google Scholar 

  36. Goldman KN, Chenette D, Arju R, Duncan FE, Keefe DL, Grifo JA, et al. mTORC1/2 inhibition preserves ovarian function and fertility during genotoxic chemotherapy. Proc Natl Acad Sci U S A. 2017;114(12):3186–91. https://doi.org/10.1073/pnas.1617233114.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Kohnoh T, Hashimoto N, Ando A, Sakamoto K, Miyazaki S, Aoyama D, et al. Hypoxia-induced modulation of PTEN activity and EMT phenotypes in lung cancers. Cancer Cell Int. 2016;16:33. https://doi.org/10.1186/s12935-016-0308-3.

    Article  PubMed  PubMed Central  Google Scholar 

  38. Drolle H, Wagner M, Vasold J, Kutt A, Deniffel C, Sotlar K, et al. Hypoxia regulates proliferation of acute myeloid leukemia and sensitivity against chemotherapy. Leuk Res. 2015;39(7):779–85. https://doi.org/10.1016/j.leukres.2015.04.019.

    Article  CAS  PubMed  Google Scholar 

  39. Di Cristofano A, Kotsi P, Peng YF, Cordon-Cardo C, Elkon KB, Pandolfi PP. Impaired Fas response and autoimmunity in Pten+/− mice. Science. 1999;285(5436):2122–5.

    Article  PubMed  Google Scholar 

  40. Fero ML, Rivkin M, Tasch M, Porter P, Carow CE, Firpo E, et al. A syndrome of multiorgan hyperplasia with features of gigantism, tumorigenesis, and female sterility in p27(Kip1)-deficient mice. Cell. 1996;85(5):733–44.

    Article  CAS  PubMed  Google Scholar 

  41. Brugarolas J, Lei K, Hurley RL, Manning BD, Reiling JH, Hafen E, et al. Regulation of mTOR function in response to hypoxia by REDD1 and the TSC1/TSC2 tumor suppressor complex. Genes Dev. 2004;18(23):2893–904. https://doi.org/10.1101/gad.1256804.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. DeYoung MP, Horak P, Sofer A, Sgroi D, Ellisen LW. Hypoxia regulates TSC1/2-mTOR signaling and tumor suppression through REDD1-mediated 14-3-3 shuttling. Genes Dev. 2008;22(2):239–51. https://doi.org/10.1101/gad.1617608.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Kong HS, Kim SK, Lee J, Youm HW, Lee JR, Suh CS, et al. Effect of exogenous anti-Mullerian hormone treatment on cryopreserved and transplanted mouse ovaries. Reprod Sci. 2016;23(1):51–60. https://doi.org/10.1177/1933719115594021.

    Article  CAS  PubMed  Google Scholar 

  44. Durlinger AL, Visser JA, Themmen AP. Regulation of ovarian function: the role of anti-Mullerian hormone. Reproduction. 2002;124(5):601–9.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Histology and Embryology, School of Medicine, Akdeniz University, Campus, 07070, Antalya, Turkey

    Soner Celik & Ciler Celik-Ozenci

  2. Department of Histology and Embryology, Ankara University School of Medicine, 06100, Ankara, Turkey

    Ferda Topal Celikkan & Alp Can

  3. Department of Obstetrics and Gynecology, Centre for Assisted Reproduction, School of Medicine, Ankara University, 06100, Ankara, Turkey

    Sinan Ozkavukcu

Authors
  1. Soner Celik
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ferda Topal Celikkan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sinan Ozkavukcu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alp Can
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ciler Celik-Ozenci
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ciler Celik-Ozenci.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Celik, S., Celikkan, F.T., Ozkavukcu, S. et al. Expression of inhibitor proteins that control primordial follicle reserve decreases in cryopreserved ovaries after autotransplantation. J Assist Reprod Genet 35, 615–626 (2018). https://doi.org/10.1007/s10815-018-1140-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-018-1140-6

Keywords

Search

Navigation