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a Laboratory for Reproductive Medicine, Marine Biological Laboratory, Woods Hole, Massachusetts 02543
b Department of Ob/Gyn, Women & Infants Hospital and Brown University, Providence, Rhode Island 02905
The oxidation of cellular sulfhydryl (SH) groups has been implicated in the induction of apoptosis in various types of cells and in the disturbance of the meiotic spindle of murine oocytes during aging. The objective of this study was to determine whether the SH-specific oxidant diamide could inhibit embryo development and induce cell death, and whether the antioxidant dithiothreitol (DTT) could counteract such effects. Exposure of mouse zygotes to diamide for 3 h at 25 or 50 µM (but not 12.5 µM) resulted in cell cycle arrest and cell death with evidence of apoptosis. At higher concentrations (100 or 200 µM), diamide induced necrosis as evidenced by propidium iodide-positive pronuclei within 24 h of treatment. Simultaneous addition of DTT at equimolar concentration prevented these effects. However, when DTT was added later, it was no longer protective. DTT also effectively protected against the thiol-oxidative damage caused by diamide in blastocysts. These results suggest that altering thiol-redox status in zygotes and blastocysts may result in cell cycle arrest, apoptosis, and/or cell death.
2 Correspondence: David Keefe, Women & Infants Hospital and Brown University, Dept. of Ob-Gyn, 101 Dudley Street, Providence, RI 02905. FAX: 401 453 7500; dkeefe{at}smtp.wihri.org
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