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a Department of Obstetrics & Gynaecology, Women & Infants Hospital, Brown University, Providence, Rhode Island 02905
b Marine Biological Laboratory, Woods Hole, Massachusetts 02543
Accumulation of reactive oxygen species during aging leads to programmed cell death (PCD) in many cell types but has not been explored in mammalian fertilized eggs, in which mitochondria are "immature," in contrast to "mature" mitochondria in somatic cells. We characterized PCD in mouse zygotes induced by either intensive (1 mM for 1.5 h) or mild (200 µM for 15 min) hydrogen peroxide (H2O2) treatment. Shortly after intensive treatment, zygotes displayed PCD, typified by cell shrinkage, cytochrome c release from mitochondria, and caspase activation, then terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining in condensed pronuclei. On the other hand, after mild treatment, zygotes arrested developmentally and showed neither cytochrome c release nor caspase activation over 48 h; until 72 h, 46% zygotes exhibited TUNEL staining, and 88% of zygotes lost plasma membrane integrity. Interestingly, mild oxidative treatment induced a decline in mitochondrial membrane potential and disruption of the mitochondrial matrix. Taken together, these results suggest that oxidative stress caused by H2O2 induces PCD in mouse zygotes and that mitochondria are involved in the early phase of oxidative stress-induced PCD. Furthermore, mitochondrial malfunction also may contribute to cell cycle arrest, followed by cell death, triggered by mild oxidative stress.
1 This work was supported in part by the National Institute of Health (NIH K081099) and Women and Infants Hospital/Brown Faculty Research Fund.
2 Correspondence: David Keefe, Dept. of Ob-Gyn, Women & Infants Hospital, Brown University, 101 Dudley Street, Providence, RI 02905. FAX: 401 453 7599; dkeefe{at}smtp.wihri.org
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