264.

HAPLOIDY BUT NOT PARTHENOGENETIC ACTIVATION LEADS TO INCREASED INCIDENCE OF APOPTOSIS IN MOUSE EMBRYOS. Liua, L., Trimarchib, J. R., Keefea, D.L. aDepartment of Obstetrics and Gynecology, Women and Infants Hospital, Brown University, Providence, RI, bLaboratory for Reproductive Medicine, Marine Biological Laboratory, Woods Hole, MA, Biology of Reproduction, 66, (1), 204-210, (2000).

Abstract:
Aneuploidy underlies failed development and possibly apoptosis of some preimplantation embryos. We employed a haploid model in the mouse to study the effects of aneuploidy on apoptosis in preimplantation embryos. Mouse metaphase II oocytes that were activated with strontium formed haploid parthenogenetic embryos with 1 pronucleus, whereas activation of oocytes with strontium plus cytochalasin D produced diploid parthenogenetic embryo controls with 2 pronuclei. Strontium induced calcium transients that mimic sperm-induced calcium oscillations, and ploidy was confirmed by chromosomal analysis. Rates of development and apoptosis were compared between haploid and diploid parthenogenetic embryos (parthenotes) and control embryos derived from in vitro fertilization (IVF). Haploid mouse parthenotes cleaved at a slower rate, and most arrested before the blastocyst stage, in contrast to diploid parthenotes or IVF embryos. Developmentally retarded haploid parthenotes exhibited apoptosis at a significantly higher frequency than did diploid parthenotes or IVF embryos. However, diploid parthenotes exhibited rates of preimplantation development and apoptosis similar to those of IVF embryos, indicating that parthenogenetic activation itself does not initiate apoptosis during preimplantation development. These results suggest that haploidy can lead to an increased incidence of apoptosis. Moreover, the initiation of apoptosis during preimplantation development does not require the paternal genome.

Link to Paper

Materials & Methods:
"Fura-loaded oocytes were washed extensively with KSOM and transferred to poly-D-lysine-coated glass-bottom dishes (MatTek Corp., Ashland, MA)."

Microscopic Technique:
Fluorescence microscopy

Cell Type(s):
Mouse oocytes,