Skip to main content

Induction of superovulation in mature mice and rats using serum of spayed female dogs


The following experiments were designed to examine the effect of serum of spayed dogs on superovulation response in mice and rats. In Experiment 1, female mice at diestrus (n=30) were divided into three equal groups and superovulated with either administration of 5 IU pregnant mare serum gonadotropin (PMSG) or recombinant follicle stimulating hormone (rFSH) (reducing dose from 2.5 to 0.5 IU) and 5 IU human chorionic gonadotropin (hCG) administered 48h later. Serum of spayed dogs was administered intraperitoneally at a reduced dose from 0.1 to 0.025 mL in a 48 h period. In Experiment 2, female rats (n=30) at diestrus stage were divided into three equal groups. Superovulation was induced using either 30 IU PMSG, or a dose reduced from 5 to 1 IU rFSH and 25 IU hCG administered 48h later. Serum of spayed dogs was administered in a reduced dose from 0.6 to 0.1 mL in a 48 hour period. Female mice and rats were mated 24 h following hCG administration. On day 14 after mating, animals were euthanized and ovarian sections were fixed for histopathological evaluation and corpus luteum (CL) counting. No significant difference observed in mean (±SEM) number of CLs between the PMSG group and the mice that received serum of spayed dog (10.4±1.3 vs 9.2±1.0). Mean (±SEM) number of CLs tended to be lower in rats that received serum of spayed dog than those of rats which received either PMSG or rFSH (15.1±1.9 vs 23.6±3.1 and 23.1±2.9, P=0.06, respectively). In conclusion, serum of spayed dogs is able to induce a superovulatory response in mice and rats.


  1. 1.

    Albers-Wolthers CH, de Gier J, Oei CH, Schaefers-Okkens AC, Kooistra HS. Validation of a noninvasive diagnostic tool to verify neuter status in dogs: The urinary FSH to creatinine ratio. Theriogenology 2016; 86(5): 1376–1381.

    CAS  Article  Google Scholar 

  2. 2.

    Beijerink NJ, Buijtels JJ, Okkens AC, Kooistra HS, Dieleman SJ. Basal and GnRH-induced secretion of FSH and LH in anestrous versus ovariectomized bitches. Theriogenology 2007; 67(5): 1039–1045.

    CAS  Article  Google Scholar 

  3. 3.

    Boland MP, Goulding D, Roche JF. Alternative gonadotrophins for superovulation in cattle. Theriogenology 1991; 35(1): 5–17.

    CAS  Article  Google Scholar 

  4. 4.

    Hasegawa A, Mochida K, Inoue H, Noda Y, Endo T, Watanabe G, Ogura A. High-Yield Superovulation in Adult Mice by Anti-Inhibin Serum Treatment Combined with Estrous Cycle Synchronization. Biol Reprod 2016; 94(1): 21.

    Article  Google Scholar 

  5. 5.

    Hoppen HO. The equine placenta and equine chorionic gonadotrophin—an overview. Exp Clin Endocrinol 1994; 102(3): 235–243.

    CAS  Article  Google Scholar 

  6. 6.

    Ishigame H, Medan MS, Kawaguchi M, Fukuda A, Watanabe G, Arai K Y, Taya K. Induction of superovulation by immuno-neutralization of endogenous inhibin in immature rats. J Reprod Dev 2005; 51(5): 559–566.

    CAS  Article  Google Scholar 

  7. 7.

    Kanitz W, Becker F, Schneider F, Kanitz E, Leiding C, Nohner H P, Pöhland R. Superovulation in cattle: practical aspects of gonadotropin treatment and insemination. Reprod Nutr Dev 2002; 42(6): 587–599.

    CAS  Article  Google Scholar 

  8. 8.

    Leão Rde B, Esteves SC. Gonadotropin therapy in assisted reproduction: an evolutionary perspective from biologics to biotech. Clinics (Sao Paulo) 2014; 69(4): 279–293.

    Article  Google Scholar 

  9. 9.

    Ludwig M, Westergaard LG, Diedrich K, Andersen C Y. Developments in drugs for ovarian stimulation. Best Pract Res Clin Obstet Gynaecol 2003; 17(2): 231–247.

    CAS  Article  Google Scholar 

  10. 10.

    Luo C, Zuñiga J, Edison E, Palla S, Dong W, Parker-Thornburg J. Superovulation strategies for 6 commonly used mouse strains. J Am Assoc Lab Anim Sci 2011; 50(4): 471–478.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. 11.

    Mukumoto S, Mori K, Ishikawa H. Efficient induction of superovulation in adult rats by PMSG and hCG. Exp Anim 1995; 44(2): 111–118.

    CAS  Article  Google Scholar 

  12. 12.

    Olson PN, Mulnix JA, Nett TM. Concentrations of luteinizing hormone and follicle-stimulating hormone in the serum of sexually intact and neutered dogs. Am J Vet Res 1992; 53(5): 762–766.

    CAS  PubMed  Google Scholar 

  13. 13.

    Reichler IM. Gonadectomy in cats and dogs: a review of risks and benefits. Reprod Domest Anim 2009; 44 Suppl 2: 29–35.

    Article  Google Scholar 

  14. 14.

    Santoro N, Randolph J. Jr. Reproductive hormones and the menopause transition. Obstet Gynecol Clin North Am 2011; 38(3): 455–466.

    Article  Google Scholar 

  15. 15.

    Takeo T, Nakagata N. Superovulation using the combined administration of inhibin antiserum and equine chorionic gonadotropin increases the number of ovulated oocytes in C57BL/ 6 female mice. PLoS One 2015; 10(5): e0128330.

    Article  Google Scholar 

  16. 16.

    Zwart-van Rijkom JE, Broekmans FJ, Leufkens H G. From HMG through purified urinary FSH preparations to recombinant FSH: a substitution study. Hum Reprod 2002; 17(4): 857–865.

    CAS  Article  Google Scholar 

Download references

Author information



Corresponding author

Correspondence to A. Mogheiseh.

Rights and permissions

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Nooranizadeh, M.H., Mogheiseh, A., Kafi, M. et al. Induction of superovulation in mature mice and rats using serum of spayed female dogs. Lab Anim Res 34, 211–215 (2018).

Download citation


  • Superovulation
  • PMSG
  • FSH
  • spayed dog serum