Journal of Assisted Reproduction and Genetics

, Volume 11, Issue 8, pp 382–388 | Cite as

Cytogenetic analysis of human oocytes parthenogenetically activated by puromycin

  • Paul De Sutter
  • Dmitri Dozortsev
  • Philippe Vrijens
  • Rita Desmet
  • Marc Dhont
Genetics

Abstract

Purpose

Treatment of aged human oocytes by puromycin allows a high rate of parthenogenetic activation and development until the first cleavage division. This technique was used for the study of the chromosome complement of oocytes which remained unfertilized after in vitro fertilization. Three hundred four unfertilized oocytes were treated with 10 Μg/ml puromycin for 6–8 hr and further cultured for 12–15 hr.

Results

Activation occurred in 90.5% of the oocytes. Heterozygous diploids with two pronuclei predominated (61%), which is in contrast to the mouse, where the majority of oocytes activated by puromycin are uniform haploids (89%).

Conclusions

Therefore we conclude that puromycin treatment induces retention of the second polar body in human oocytes, unlike in mouse oocytes treated in the same way. Chromosome analysis performed on 182 oocytes suggested a nondisjunction (ND) rate for the second meiotic division of 12.7%. This is a low figure considering the fact that puromycin itself has been reported to induce nondisjunction. For the first meiotic division a ND rate of only 5.6% was found. This rate is lower than the one found in metaphase II arrested oocytes and we believe that this difference is due to the technical differences between the study of meiotic and that of mitotic chromosomes.

Key words

chromosomes human oocytes parthenogenesis puromycin 

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References

  1. 1.
    Abramczuk JW, Lopata A: Resistance of human follicular oocytes to parthenogenetic activation: DNA distribution and content in oocytes maintained in vitro. Hum Reprod 1990; 5:578–581PubMedGoogle Scholar
  2. 2.
    Winston N, Johnson M, Pickering S, Braude P: Parthenogenetic activation and development of fresh and aged human oocytes. Fertil Steril 1991;56:904–912PubMedGoogle Scholar
  3. 3.
    Balakier H, Casper RF: Experimentally induced parthenogenetic activation of human oocytes. Hum Reprod 1993;8:740–743PubMedGoogle Scholar
  4. 4.
    De Sutter P, Dozortsev D, Cieslak J, Wolf G, Verlinsky Y, Dyban A: Parthenogenetic activation of human oocytes by puromycin. J Assist Reprod Genet 1992;9:328–337PubMedGoogle Scholar
  5. 5.
    De Sutter P, Dhont M, Vanluchene E, Vandekerckhove D: Correlations between follicular fluid steroid analysis and maturity and cytogenetic analysis of human oocytes that remained unfertilized after in vitro fertilization. Fertil Steril 1991;55:958–963PubMedGoogle Scholar
  6. 6.
    Dyban AP: An improved method for chromosome preparations from preimplantation mammalian embryos, oocytes or isolated blastomeres. Stain Technol 1983;58:69–72PubMedGoogle Scholar
  7. 7.
    Siracusa G, Whittingham DG, Molinaro M, Vivarelli E: Parthenogenetic activation of mouse oocytes induced by inhibitors of protein synthesis. J Embryol Exp Morphol 1978;43:157–166PubMedGoogle Scholar
  8. 8.
    Clarke HJ, Masui Y: The induction of reversible and irreversible chromosome decondensation by protein synthesis inhibition during meiotic maturation of mouse oocytes. Dev Biol 1983;97:291–301PubMedGoogle Scholar
  9. 9.
    De Sutter P: Non-random chromosome nondisjunction at the first meiotic division induced by puromycin in the mouse.In abstract book of the IXth Meeting of the Contact Group for IVF, National Scientific Research Foundation, Brussels, 1993, p 15Google Scholar
  10. 10.
    Pellestor F: Frequency and distribution of aneuploidy in human female gametes. Hum Genet 1991;86:283–288PubMedGoogle Scholar
  11. 11.
    Angell RR: Predivision in human oocytes failing to fertilise after insemination in vitro. Hum Genet 1991;86:383–387PubMedGoogle Scholar
  12. 12.
    Angell RR, Xian J, Keith J: Chromosome anomalies in human oocytes in relation to age. Hum Reprod 1993;8:1047–1054PubMedGoogle Scholar
  13. 13.
    Kamiguchi Y, Rosenbusch B, Sterzik K, Mikamo K: Chromosomal analysis of unfertilized human oocytes prepared by a gradual fixation-air drying method. Hum Genet 1993;90:533–541PubMedGoogle Scholar
  14. 14.
    Kaufman MH: The ovulation and activation of primary and secondary oocytes in Lt/Sv strain mice. Gamete Res 1986; 14:255–264Google Scholar
  15. 15.
    Schmiady H, Sperling K, Kentenich H, Stauber M: Prematurely condensed human sperm chromosomes after in vitro fertilization (IVF). Hum Genet 1986;74:441–443PubMedGoogle Scholar
  16. 16.
    Schmiady H, Kentenich H: Premature chromosome condensation after in-vitro fertilization. Hum Reprod 1989;4:689–695PubMedGoogle Scholar
  17. 17.
    Balakier H, Casper RF: A morphologic study of unfertilized oocytes and abnormal embryos in human in vitro fertilization. J Vitro Fert Embryo Transfer 1991;8:73–79Google Scholar
  18. 18.
    Dyban AP, Noniashvili EM: Parthenogenetic development of mouse eggs activated by heat shock. Ontogenez 1986;17:587–598PubMedGoogle Scholar
  19. 19.
    Procházka R, Kaňka J, šutovský P, Fulka J, Motlík J: Development of pronuclei in pig oocytes activated by a single electric pulse. J Reprod Fert 1992;96:725–734Google Scholar
  20. 20.
    Balakier H, Tarkowski: Diploid parthenogenetic mouse embryos produced by heat shock and cytochalasin B. J Embryol Exp Morphol 1976;35:25–39PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1994

Authors and Affiliations

  • Paul De Sutter
    • 1
  • Dmitri Dozortsev
    • 1
  • Philippe Vrijens
    • 1
  • Rita Desmet
    • 1
  • Marc Dhont
    • 1
  1. 1.Center for Infertility, Department of Obstetrics/GynecologyUniversity HospitalGentBelgium

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