Journal of Assisted Reproduction and Genetics

, Volume 30, Issue 11, pp 1421–1429 | Cite as

Serum markers of ovarian reserve and ovarian histology in adult mice treated with cyclophosphamide in pre-pubertal age

  • Laura DettiEmail author
  • Rebecca A. Uhlmann
  • Meifen Lu
  • Michael P. Diamond
  • Ghassan M. Saed
  • Nicole M. Fletcher
  • Jie Zhang
  • Lucy J. Williams
Fertility Preservation



AMH is used to quantify the extent of follicular pool in postpubertal women, but its value after chemotherapy is unclear. We tested AMH as a marker of follicular reserve in adult mice treated with cyclophosphamide (CTX) in prepubertal age.


Mice received placebo or CTX at age 18 days. AMH and FSH were assessed on day 43, 56, and 95 of life. Ovaries were fixed in formalin, embedded in paraffin, and stained with H&E and TUNEL. Follicular apoptosis was graded.


All mice exposed to CTX had a decreased number of follicles/mm2 and significantly decreased AMH, but only 48 % of pubertal and 81 % of adult mice had increased FSH. Over time, there was an increase in FSH (p < 0.05), but not a concurrent decrease in AMH, while in controls, FSH remained stable and AMH decreased. There was no correlation between histological and serological markers.


CTX administration to pre-pubertal mice caused various degrees of residual function, which were reflected by FSH, but not by AMH or by the number of ovarian follicles. AMH served as a marker of quantitative, and FSH of qualitative, residual ovarian function.


Animal study Prepubertal Cyclophosphamide AMH FSH Follicle count 



The study was funded with an institutional grant from the University of Tennessee Health Science Center to Dr. Laura Detti.


  1. 1.
    Bath LE, Wallace WH, Shaw MP, Fitzpatrick C, Anderson RA. Depletion of ovarian reserve in young women after treatment for cancer in childhood: detection by anti-Mullerian hormone, inhibin B and ovarian ultrasound. Hum Reprod. 2003;18:2368–74.PubMedCrossRefGoogle Scholar
  2. 2.
    Browne HN, Moon KS, Mumford SL, et al. Is anti-Müllerian hormone a marker of acute cyclophosphamide-induced ovarian follicular destruction in mice pretreated with cetrorelix? Fertil Steril. 2011;96:180–6.PubMedCrossRefGoogle Scholar
  3. 3.
    Chemaitilly W, Mertens AC, Mitby P, et al. Acute ovarian failure in the childhood cancer survivor study. J Clin Endocrinol Metab. 2006;91:1723–8.PubMedCrossRefGoogle Scholar
  4. 4.
    de Bruin JP, Dorland M, Spek ER, et al. Ultrastructure of the resting ovarian follicle pool in healthy young women. Biol Reprod. 2002;66:1151–60.PubMedCrossRefGoogle Scholar
  5. 5.
    de Vet A, Laven JS, de Jong FH, Themmen APN, Fauser BC. Antimullerian hormone serum levels: a putative marker for ovarian aging. Fertil Steril. 2002;77:357–62.PubMedCrossRefGoogle Scholar
  6. 6.
    Decanter C, Morschhauser F, Pigny P, Lefebvre C, Gallo C, Dewailly D. Anti-Müllerian hormone follow-up in young women treated by chemotherapy for lymphoma: preliminary results. Reprod Biomed Online. 2010;20:280–5.PubMedCrossRefGoogle Scholar
  7. 7.
    Detti L, Martin DC, Williams RD, Schlabritz-Loutsevich N, Williams LJ, Uhlmann RA. Somatic and reproductive outcomes in mice treated with cyclophosphamide in pre-pubertal age. Syst Biol Reprod Med. 2013;59:140–5.PubMedCrossRefGoogle Scholar
  8. 8.
    Di Clemente N, Goxe B, Remy JJ, et al. Inhibitory effect of AMH upon aromatase activity and LH receptors of granulosa cells of rat and porcine immature ovaries. Endocrine. 1994;2:553–8.Google Scholar
  9. 9.
    Durlinger ALL, Kramer P, Karels B, et al. Control of primordial follicle recruitment by anti-Mullerian hormone in the mouse ovary. Endocrinol. 1999;140:5789–96.CrossRefGoogle Scholar
  10. 10.
    Durlinger ALL, Visser JA, Themmen APN. Regulation of ovarian function: the role of anti-Mullerian hormone. Reprod. 2002;124:601–9.CrossRefGoogle Scholar
  11. 11.
    Fanchin R, Schonauer LM, Righini C, Guibourdenche J, Frydman R, Taieb J. Serum anti-Mullerian hormone is more strongly related to ovarian follicular status than serum inhibin B, estradiol, FSH and LH on day 3. Hum Reprod. 2003;18:323–7.PubMedCrossRefGoogle Scholar
  12. 12.
    Green DM, Sklar CA, Boice Jr JD, et al. Ovarian failure and reproductive outcomes after childhood cancer treatment: results from the Childhood Cancer Survivor Study. J Clin Oncol. 2009;27:2374–81.PubMedCrossRefGoogle Scholar
  13. 13.
    Kevenaar ME, Meerasahib MF, Kramer P, et al. Serum anti-mullerian hormone levels reflect the size of the primordial follicle pool in mice. Endocrinol. 2006;147:3228–34.CrossRefGoogle Scholar
  14. 14.
    Kline JK, Kinney AM, Levin B, Kelly AC, Ferin M, Warburton D. Trisomic pregnancy and elevated FSH: implications for the oocyte pool hypothesis. Hum Reprod. 2011;26:1537–50.PubMedCrossRefGoogle Scholar
  15. 15.
    Meirow D, Nugent D. (2001) The effects of radiotherapy and chemotherapy on female reproduction. Hum Reprod Update. 2001;7:535–43.PubMedCrossRefGoogle Scholar
  16. 16.
    Ohnemus U, Unalan M, Handjiski B, Paus R. Topical estrogen accelerates hair regrowth in mice after chemotherapy-induced alopecia by favoring the dystrophic catagen response pathway to damage. J Invest Dermatol. 2004;122:7–13.PubMedCrossRefGoogle Scholar
  17. 17.
    Oktay K, Briggs D, Gosden RG. Ontogeny of follicle-stimulating hormone receptor gene expression in isolated human ovarian follicles. J Clin Endocrinol Metab. 1997;82:3748–51.PubMedCrossRefGoogle Scholar
  18. 18.
    Oktem O, Oktay K. Quantitative assessment of the impact of chemotherapy on ovarian follicle reserve and stromal function. Cancer. 2007;110:2222–9.PubMedCrossRefGoogle Scholar
  19. 19.
    Oktem O, Oktay K. A novel ovarian xenografting model to characterize the impact of chemotherapy agents on human primordial follicle reserve. Cancer Res. 2007;67:10159–62.PubMedCrossRefGoogle Scholar
  20. 20.
    Pepling ME, Spradling AC. Mouse ovarian germ cell cysts undergo programmed breakdown to form primordial follicles. Dev Biol. 2001;234:339–51.PubMedCrossRefGoogle Scholar
  21. 21.
    Sahambi SK, Visser JA, Themmen AP, Mayer LP, Devine PJ. Correlation of serum anti-Mullerian hormone with accelerated follicle loss following 4-vinyl-cyclohexene diepoxide-induced follicle loss in mice. Reprod Toxicol. 2008;26:116–22.PubMedCrossRefGoogle Scholar
  22. 22.
    Sanders JE, Hawley J, Levy W, Gooley T, Buckner CD, Deeg HJ, et al. Pregnancies following high-dose cyclophosphamide with or without high-dose busulfan or total-body irradiation and bone marrow transplantation. Blood. 1996;87:3045–52.PubMedGoogle Scholar
  23. 23.
    Sarafoglou K, Boulad F, Gillio A, Sklar C. Gonadal function after bone marrow transplantation for acute leukemia during childhood. J Pediatr. 1997;130:210–6.PubMedCrossRefGoogle Scholar
  24. 24.
    Tingen CM, Bristol-Gould SK, Kiesewetter SE, Wellington JT, Shea L, Woodruff TK. Prepubertal primordial follicle loss in mice is not due to classical apoptotic pathways. Biol Reprod. 2009;81:16–25.PubMedCrossRefGoogle Scholar
  25. 25.
    Vaskivuo TE, Anttonen M, Herva R, et al. Survival of human ovarian follicles from fetal to adult life: apoptosis, apoptosis-related proteins, and transcription factor GATA-4. J Clin Endocrinol Metab. 2001;86:3421–9.PubMedCrossRefGoogle Scholar
  26. 26.
    Vialard F, Boitrelle F, Molina-Gomes D, Selva J. Predisposition to aneuploidy in the oocyte. Cytogenet Genome Res. 2011;133:127–35.PubMedCrossRefGoogle Scholar
  27. 27.
    Visser JA, de Jong FH, Laven JS, Themmen AP. Anti-Müllerian hormone: a new marker for ovarian function. Reprod. 2006;131:1–9.CrossRefGoogle Scholar
  28. 28.
    Wallace WH, Thomson AB, Saran F, Kelsey TW. Predicting age of ovarian failure after radiation to a field that includes the ovaries. Int J Radiat Oncol Biol Phys. 2005;62:738–44.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Laura Detti
    • 1
    Email author
  • Rebecca A. Uhlmann
    • 1
  • Meifen Lu
    • 2
  • Michael P. Diamond
    • 3
  • Ghassan M. Saed
    • 4
  • Nicole M. Fletcher
    • 4
  • Jie Zhang
    • 2
  • Lucy J. Williams
    • 1
  1. 1.Department of Obstetrics and GynecologyUniversity of Tennessee Health Science CenterMemphisUSA
  2. 2.Department of PathologyMemphisUSA
  3. 3.Georgia Regents UniversityDepartment of Obstetrics and GynecologyAugustaUSA
  4. 4.Department of Obstetrics and GynecologyWayne State UniversityDetroitUSA

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