Advertisement

Optimization of superovulation in the reproductively mature mouse

  • Michael Legge
  • Martin H. Sellens
Animal Investigations

Abstract

Optimum gonadotropin doses and chronology were established for the induction of superovulation in sexually mature hybrid mice (BALB/cBy×C57BL/6By). A regime of 12 IU pregnant mares' serum gonadotropin (PMSG), followed 48 hr later by 20 IU human chorionic gonadotropin (hCG) administered 1 hr before the midpoint of the light cycle (1200), gave the maximum ovulatory response. There was no evidence that endogenous luteinizing hormone influenced the superovulation response to exogenous gonadotropins. Fewer than 50% of zygotes reached the blastocyst stage (90–93 hr post hCG), with the greatest rate of loss at the two-to four-cell stage. Litter size following superovulation was 19.6±0.9. There was no significant difference between the number of blastocysts observed and litter size. Similarly, counts of mature follicles in ovaries prior to hCG stimulation were not significantly greater than the number of secondary oocytes that subsequently ovulated. These data indicate that standard superovulation protocols may require finetuning to maximize productivity and confirm that embryo loss is greatest between the first cleavage division and blastocyst formation.

Key words

superovulation mouse optimization oocytes blastocysts ovaries 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Runner MN, Gates AH: Conception in prepubertal mice following artificially induced ovulation and mating. Nature 1954;174:222–223Google Scholar
  2. 2.
    Fowler RE, Edwards RE. Androgen production by theca and granulosa cells isolated from proestrous rat follicles. Endocrinology 1957;100:1341–1347Google Scholar
  3. 3.
    Edwards RG, Gates AH: Timing of the stages of the maturation divisions, ovulation, fertilization and the first cleavage of eggs in adult mice treated with gonadotrophins. J Endocrinol 1959;21:292–324Google Scholar
  4. 4.
    Edwards RG, Fowler RE: Superovulation treatment of adult mice. Their subsequent natural fertility and response to further treatment. J Endocrinol 1960;21:147–154Google Scholar
  5. 5.
    Miller BG, Armstrong DT: Effects of a superovulating dose of pregnant mares serum gonadotrophin in ovarian function serum oestradiol and progesterone levels and early embryo development in immature rats. Biol Reprod 1981;25:261–271PubMedGoogle Scholar
  6. 6.
    Gates AH: Time relationship between neural stimulus for LH-releasing factor and ovulation in the PMS treated mouse. Am Zool 1969;9:1080Google Scholar
  7. 7.
    Allen J, McLaren A: Cleavage rate of mouse eggs from induced and spontaneous ovulation. J Reprod Fert 1971;27:137–140Google Scholar
  8. 8.
    Beaumont HM, Smith AP: Embryonic mortality during pre and post-implantation periods of pregnancy in mature mice after superovulation. J Reprod Fert 1975;67:281–284Google Scholar
  9. 9.
    Spindle AI, Goldstein LS: Induced ovulation in mature mice and developmental capacity of the embryos in-vitro. J Reprod Fert 1975;44:113–116Google Scholar
  10. 10.
    Champlin AK, Dorr DL, Gates AH: Determining the stage of the oestrous cycle in the mouse by appearance of the vagina. Biol Reprod 1973;8:491–494PubMedGoogle Scholar
  11. 11.
    Allen E: The oestrous cycle in the mouse. Am J Anat 1922;30:297–371CrossRefGoogle Scholar
  12. 12.
    Snell GD, Fekete E, Hummel KP, Abelmann WH: The relation of mating, ovulation and the oestrous smear in the house mouse to time of day. Anat Rec 1940;76:39–54CrossRefGoogle Scholar
  13. 13.
    Hogan B, Constantani F, Lacy E:In Manipulating the Mouse Embryo. Cold Spring Harbor, NY, Cold Spring Harbor Laboratory, 1986Google Scholar
  14. 14.
    Champlin AK, Kusia SJ, Rice BA, Mobraaten LE: Cell surface characteristics of blastocysts from spontaneously ovulating and gonadotrophin treated mice. Biol Reprod 1987;36:439–444PubMedGoogle Scholar
  15. 15.
    Sasamoto S, Sato K, Naito M: Biological active life of PMSG in mice with special reference to follicular ability to ovulate. J Reprod Fert 1972;59:353–363Google Scholar
  16. 16.
    Hetherington CM: Mouse husbandry.In Mammalian Development: A Practical Approach, M Monk (ed). Oxford, IRL Press, 1987, pp 1–12Google Scholar
  17. 17.
    Bingel AS, Schwartz NB: Pituitary LH content and reproductive tract changes during mouse oestrous cycle. J Reprod Fert 1969;19:215–222Google Scholar
  18. 18.
    Bingel AS, Schwartz NB: Timing of LH release and ovulation in the cyclic mouse. J Reprod Fert 1969;19:223–229Google Scholar
  19. 19.
    Bingel AS, Schwartz NB: Timing of LH release and ovulation in the post-partum mouse. J Reprod Fert 1969;19:231–237Google Scholar
  20. 20.
    Ryle M: The time factor in responses to pituitary gonadotrophins by mouse ovaries in-vitro. J Reprod Fert 1971;25:61–74Google Scholar
  21. 21.
    Murr SM, Geschwind II, Bradford GE: Plasma LH and FSH during different oestrous cycle conditions in mice. J Reprod Fert 1973;32:221–230Google Scholar
  22. 22.
    Flurkey K, Gee DM, Sinha YN, Wisner JR, Finch CE: Age effects of luteinizing hormone, progesterone and prolactin in proestrous and acyclic C57BL/6J mice. Biol Reprod 1982;26:835–846PubMedGoogle Scholar
  23. 23.
    Miller BG, Armstrong DT: Superovulatory doses of pregnant mares serum gonadotrophin cause delayed implantation and infertility in immature rats. Biol Reprod 1981;25:253–260PubMedGoogle Scholar
  24. 24.
    Pedersen T: Follicle kinetics in the ovary of the cycling mouse. Acta Endocrinol 1970;64:304–323PubMedGoogle Scholar
  25. 25.
    Pedersen T: Follicle growth in the mouse ovary.In: Oogenesis, JD Biggers, AW Schuetz (eds). Baltimore: University Park Press, 1972, pp 361–376Google Scholar
  26. 26.
    McNatty KP, Hunter WM, McNeilly AS, Sawers RS: Changes in the concentration of pituitary and steroid hormones in the follicular fluid of human graafian follicles throughout the menstrual cycle. J Endocrinol 1975;64:555–571PubMedGoogle Scholar
  27. 27.
    Byskov AGS: Atresia.In Ovarian Follicular Development and Function, AR Midgley, WA Sadler (eds). New York: Plenum Press, 1978, pp 41–58Google Scholar
  28. 28.
    Rousseau P, Meda P, Lecart C: Cortical granule release in human follicular oocytes. Biol Reprod 1977;16:104–111PubMedGoogle Scholar
  29. 29.
    Hyttel P, Westergaard L, Byskov AG: Ultrastructure of human cumulus-oocyte complexes from healthy and atretic follicles. Hum Reprod 1986;1:153–157PubMedGoogle Scholar
  30. 30.
    Oakberg E: Follicular growth and atresia in the mouse. In Vitro 1979;15:41–49PubMedGoogle Scholar
  31. 31.
    Byskov AGS: Cell kinetics of follicular atresia in the mouse ovary. J Reprod Fert 1974;37:277–285Google Scholar
  32. 32.
    Eppig JJ: FSH stimulates hyaluronic acid synthesis by oocyte-cumulus cell-cell complexes from mouse preovulatory follicles. Nature 1979;281:483–484CrossRefPubMedGoogle Scholar
  33. 33.
    Salustri A, Siracussa G: Metabolic coupling, cumulus expansion and meiotic resumption in mouse cumuli oophari cultured in-vitro in the presence of FSH or dcAMP, or stimulated in vivo by hCG. J Reprod Fert 1983;68:335–341Google Scholar
  34. 34.
    Warner CM, Brownell MS, Ewoldsen MA: Why aren't embryos immunologically rejected by their mothers? Biol Reprod 1988;39:17–29Google Scholar

Copyright information

© Plenum Publishing Corporation 1994

Authors and Affiliations

  • Michael Legge
    • 1
  • Martin H. Sellens
    • 1
  1. 1.Department of BiologyUniversity of EssexColchesterEngland

Personalised recommendations