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单细胞小鼠胚胎与第二极体染色体融合后的可视化及细胞遗传学分析。

Visualization and cytogenetic analysis of second polar body chromosomes following its fusion with a one-cell mouse embryo.

作者信息

Verlinsky Y, Dozortsev D, Evsikov S

机构信息

Reproductive Genetics Institute, Chicago, Illinois 60657.

出版信息

J Assist Reprod Genet. 1994 Mar;11(3):123-31. doi: 10.1007/BF02332089.

Abstract

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.

摘要

目的

本研究旨在通过将第二极体(2PB)与单细胞期小鼠胚胎进行电融合,来可视化2PB染色体,从而实现染色体疾病的孕前诊断。

结果

通过2PB与小鼠受精卵的电融合,已实现80%至90%的杂交效率。在40%至50%的杂交体中可观察到2PB染色体。当与通过显微手术从单细胞胚胎中去除原核而获得的胞质体融合时,65%的2PB染色体得以可视化。这些所得中期相中多达33%至43%似乎含有染色体畸变。对体外重建的单细胞期杂交体发育的后续观察显示其活力显著下降。由2PB与去核受精卵电融合产生的杂交胚胎在两细胞期之后不再发育。

结论

电融合是2PB与单细胞小鼠胚胎杂交的有效方法,可能有助于2PB染色体的可视化和细胞遗传学分析。如果2PB与去核受精卵融合,2PB染色体的可视化率会更高。然而,该方法会诱导超过30%的染色体畸变,并可能导致所得单细胞胚胎的活力显著下降。

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