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Visualization and cytogenetic analysis of second polar body chromosomes following its fusion with a one-cell mouse
Y Verlinsky1, D Dozortsev, S Evsikov
1Reproductive Genetics Institute, Chicago, Illinois 60657.
Purpose:
This study was designed to visualize the second polar body (2PB) chromosomes using its electrofusion with a one-cell-stage mouse embryo to approach preconception diagnosis of chromosomal disorders.
Results:
Eighty to 90% hybridization efficiency has been achieved by electrofusion of 2PB with mouse zygotes. 2PB chromosomes were visualized in 40-50% of hybrids. Sixty-five percent of 2PB chromosomes were visualized when fused with the cytoplast obtained microsurgically by removing pronuclei from a one-cell embryo. As much as 33-43% of these resulting metaphases appeared to contain chromosomal aberrations. The follow-up of the development of the reconstructed one cell-stage hybrids in vitro revealed a significant decrease in their viability. The hybrid embryos resulting from 2PB electrofusion with enucleated zygotes did not develop beyond the two-cell stage.
Conclusion:
Electrofusion is an efficient approach for hybridization of 2PB with a one-cell mouse embryo and may be useful for visualization and cytogenetic analysis of 2PB chromosomes. The visualization rate of 2PB chromosomes is higher if 2PB is fused with enucleated zygotes. However, the method induces over 30% of chromosomal aberrations and may lead to a significant decrease in the viability of the resulting one-cell embryos.