Molecular Human Reproduction, Vol 4, 775-783, Copyright © 1998 by Oxford University Press
R Levy, M Benchaib, H Cordonier, C Souchier and JF Guerin
Confocal laser scanning microscopy was used to observe human arrested and
fragmented preimplantation embryos obtained by in-vitro fertilization.
Observation of the cellular actin cortex and chromatin showed a high
frequency of embryos with blastomeres exhibiting two or more nuclei, while
others had nuclei displaying chromatin condensation and fragmentation
patterns. Many of the abnormal chromatin images could be due to the process
of programmed cell death (apoptosis). The possible link between
abnormalities of the blastomeres and apoptosis was investigated using two
detection methods for cells undergoing apoptosis. Detection of
phosphatidylserine exposure was performed using annexin V; the chromosomal
breakdown preceding the nuclear collapse of apoptotic nuclei was tested
using the terminal transferase-mediated DNA end labelling (TUNEL) assay.
Annexin V staining was observed in all arrested and/or fragmented human
embryos, but not in cryopreserved embryos which continued to develop
normally after thawing. The TUNEL assay was positive in 30% (15/50) of
arrested embryos, all of which had cytoplasmic fragments. In contrast,
embryos showing regular size blastomeres without fragments were TUNEL
negative.
JOURNAL ARTICLE
Annexin V labelling and terminal transferase-mediated DNA end labelling (TUNEL) assay in human arrested embryos
Laboratoire de Biologie de la Reproduction et du Developpement, Hopital Edouard Herriot, Lyon, France.
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