Hostname: page-component-848d4c4894-hfldf Total loading time: 0 Render date: 2024-05-31T12:08:49.219Z Has data issue: false hasContentIssue false
  • English
  • Français

The parental origin of the distal pronucleus in dispermic human zygotes

Published online by Cambridge University Press:  26 September 2008

Ya-xu Tang ,
Santiago Munné ,
Adrienne Reing ,
Glenn Schattman ,
Jamie Grifo  and
Jacques Cohen
Ya-xu Tang
Affiliation:
The Center for Reproductive Medicine and Infertility, Departement of Obstetrics and Gynecology, Cornell University Medical College, New york, NY 10021, USA.
Santiago Munné
Affiliation:
The Center for Reproductive Medicine and Infertility, Departement of Obstetrics and Gynecology, Cornell University Medical College, New york, NY 10021, USA.
Adrienne Reing
Affiliation:
The Center for Reproductive Medicine and Infertility, Departement of Obstetrics and Gynecology, Cornell University Medical College, New york, NY 10021, USA.
Glenn Schattman
Affiliation:
The Center for Reproductive Medicine and Infertility, Departement of Obstetrics and Gynecology, Cornell University Medical College, New york, NY 10021, USA.
Jamie Grifo
Affiliation:
The Center for Reproductive Medicine and Infertility, Departement of Obstetrics and Gynecology, Cornell University Medical College, New york, NY 10021, USA.
Jacques Cohen*
Affiliation:
The Center for Reproductive Medicine and Infertility, Departement of Obstetrics and Gynecology, Cornell University Medical College, New york, NY 10021, USA.
*
J. Cohen, The Gamete and Embryo Research Laboratory, 1300 York Avenue, P.O. Box 30, New York, NY 10021, USA. Telephone: 212-7466626 Fax: 212-7468589.
Get access Rights & Permissions [Opens in a new window]

Summary

The purpose of this investigation was to determine the parental origin og the pronucleus furthest from the second polar body (the distal pronucleus) in dispermic human zygotes. Infact dispermic embryos (n = 53) and those from which the distal pronucles (n =50) was removed at the zygote stage were biopsied after cleavage. Blastomeres were sexed using either coamplification of X and Y probes using a duplex polymerase chain reaction (PCR), or simultaneous fluorescence in situ hybridisation (FISH) with directly fluorochrome-labelled probes for chromosomes X, Y and 18. The ratio X/Y was determined in both groups of embryos by assessing a minimum of two blastomeres. If the pronuclei in dispermic zygotes are topographcially in a fixed position, the X/Y ratio should change from 1:3 in dispermic embryos to 1:1 in enucleated ones. The ratio of embryos containing only an X chromosome and those with X as well as Y chromosomes in the intact dispermic zygotes was 1.0:2.3 which is similar to the theoretical ratio of 1:3. This ratio was 1.0:1.3 in dispermic zygotes from which the distal pronuclei were removed. This ratio is not significantly different from the 1:1 ratio based on a statistical analysis with a sample size of 50. These sex ratios would have been considered different if more than 200 enucleations had been performed. Although the ratio X/Y was altered following removal of distal pronuclei, suggesting frequent targeting of male pronuclei, accidental removal of the female pronucleus could not be excluded. This indicates that enucleation of dispermic zygotes could produce high yields of gynogenetic and androgenetic embryos for research purposes. Clinical application aimed at producing biparental zygotes may be hazardous, since mosaicism was common among enucleated embryos.

Keywords

DispermyEnucleationFISHPCRPronuclear position
Type
Commentary
Information
Zygote , Volume 2 , Issue 1 , February 1994 , pp. 79 - 85
Copyright
Copyright © Cambridge University Press 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Balakier, H. & Czolowska, R. (1977). Cytoplasmic control of nuclear maturation in mouse oocytes. Exp. Cell Res. 112, 137–41.Google Scholar
Borsuk, E. (1991). Anucleate fragments of parthenogenetic eggs and of maturing oocytes contain complementary factors required for development of a male pronucleus. Mol. Reprod.Dev. 29, 150–6Google Scholar
Clarke, H.J. & Masui, Y. (1986). Transformation of sperm nuclei to metaphase chromosomes in the cytoplasm of maturing oocytes of the mouse. J. Cell Biol. 102, 1039–46.CrossRefGoogle ScholarPubMed
Clarke, H.J. & Masui, Y. (1987). Dose-dependent relationship between oocyte cytoplasmic volume and transformation of sperm nuclei to metaphase chormosomes, J.Cell. Biol. 104, 831–40.Google Scholar
Cohen, J. (1993). Micromanipulation of human gametes, zygotes and embryos. In Infertility: Evaluation and Treatment, ed, Keye Change, Rebar & Soules, . Philadelphia: W.B Saunders (in press).Google Scholar
Cohen, J., Malter, H.E., Talansky, B.E. & Grifo, J. (1992). Micromanipulation of Human Gametes and Embryos. New York: Raven Press.Google ScholarPubMed
Gordon, J.W. & Talansky, B.E. (1986). Assisted fertilization by zona drilling: a mouse model for correction of oligospermia. J. Exp. Zool. 239, 347–54.CrossRefGoogle Scholar
Gordon, J.W., Grunfeld, L., Garrisi, G.J.,Navot, D. & Laufer, N. (1989). Successful microsurgical removal of a pronucleus from tripronuclear human zygotes. Fretil. Steril. 52, 367–72.Google Scholar
Grifo, J.A., Tang, Y.X., Kogelman, L., Pratten, M.K., Sanyal, M.K. & Fenton, W. (1990). Characterization of a new human Y-chromosome specific primer pair for polymerase chain reaction. J. In Vitro Fertil. Embryo Transfer 7, 192, abstract 17.Google Scholar
Grifo, J.A., Tang, Y.X., Cohen, J., Gilbert, F., Sanyal, M.K. &Rosenwaks, Z. (1992).Pregnancy after embryo biopsy and coamplification of DNA from X and Y chromosomes from single blastomers. J. Am. Med. Assoc. 6, 727–9.CrossRefGoogle Scholar
Hopman, A.H.N., Ramaekers, F.C.S., Raap, A.K., Beck, J.L.M., Devilee, P., van der Ploeg, M.. & Voojis, G.P. (1988). In situ hybridizations in solid bladder tumors. Histochemistry 89, 307–16.Google Scholar
Johnson, J.D. & Nogueira, Araujo GM. (1981). A simple method of reducing the fading of immunofluorescence during microscopy. J. Imunol. Methods 43, 349–51.Google Scholar
Kajii, T. & Solter, D. (1983). Nuclear transplantation in the mouse embryo by microsurgery and cell fusion. Science 220, 1300–2.Google Scholar
Malter, H.E.& Cohen, J. (1989). Embryonic development after microsurgical repair of polyspermic human zygotes. Fertil. Steril. 52, 373–80.CrossRefGoogle ScholarPubMed
Masui, Y. (1985). Problems of oocyte maturation and the control of chromosome cycles. Dev. Growth Differ. 27, 295–309.CrossRefGoogle ScholarPubMed
Moore, D.H. & Gledhill, B.L. (1988). How large should my study be so that I can detect an altered sex ratio?. Fertil. Steril. 50, 21–5.CrossRefGoogle ScholarPubMed
Munné, s., Weier, H.U.G., Stein, J., Grifo, J. & Cohen, J. (1993). A fast and efficient method for simultaneous X and Y in situ hybridization of human blastomeres. J. Assist. Reprod. Genet. 10, 82–90.Google Scholar
Pieters, M.H.E.C., Dumoulin, J.C.M., Geraedits, J.P.M. (1992). Triploidy after in vitro fertilization: cytogenetic analysis of human zygotes and embryos. J. Assisted Reprod. Genet. 9, 68–76.Google Scholar
Quinn, I., Warnes, G.M., Kerin, J.F. & Kirby, C. (1985). Culture factors affecting the success rate of IVF and embryo transfer. Ann NY Acad. Sci. 442, 266–72.Google Scholar
Santella, L., Alikani, M., Talansky, B.E., Cohen, J. & Dale, B. (1992). Is the human oocyte plasma membrane polarized?. Hum. Reprod. 7,999–1003.CrossRefGoogle ScholarPubMed
Surani, M.A.H.. & Barton, S.C. (1983). Development of gynogenetic egss in the mouse: implications for parthenogenetic embryos. Science 222, 1034–6.CrossRefGoogle Scholar
Tarkowski, A.K. (1966). An air drying method for chromosome preparation from eggs. Cyclogenetics 5, 394–400.CrossRefGoogle Scholar
Waye, J.S. & Willard, H.F. (1985). Chromosome specific alpha satellite DNA: nucleotide sequence of the 2.0 kilobase pair repeat from the human X chromosome. Nucleic Acids Res. 13, 2731–43.Google Scholar
Wiker, H., Wright, G. & Cohen, J. (1990). Recognition of paternal pronuclei in human zygotes. J. In Vitro Fertil. Embryo Transfer 7, 33–7.Google Scholar
Willadsen, S. (1986). Nuclear transplantation in sheep embryos. Nature 320, 63–5.Google Scholar
Wolf, D.P. (1981). The mammalian egg's block to polyspermy. In Fertilization and Embryonic Development In Vitro, ed. Nastroianni, L. Jr & Biggers, JD, pp. 183–97. New York: Plenum Press.CrossRefGoogle Scholar