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The parental origin of the distal pronucleus in dispermic human zygotes

Published online by Cambridge University Press:  26 September 2008

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.

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.

Type
Commentary
Copyright
Copyright © Cambridge University Press 1994

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