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© 2001 by the American Diabetes Association, Inc.
Expression and Role of Bcl-2 in Rat Blastocysts Exposed to High D-Glucose
From the Physiology of Human Reproduction Research Unit (S.P., S.C., I.V., R.D.H.), Université Catholique de Louvain, Brussels; the Cell Biology Unit (P.V.D.S., P.J.C.), Christian de Duve Institute of Cellular Pathology, Brussels, Belgium; the Division of Biology and Embryology (A.V.C., H.A.), Université de Mons-Hainaut, Mons and the Veterinary Unit (I.D.), Université Catholique de Louvain, Louvain-la-Neuve, Belgium.
Address correspondence and reprint requests to Serge Pampfer, PhD, OBST 5330 Research Unit, University of Louvain Medical School, 53 Avenue Mounier, 1200 Brussels, Belgium. E-mail: pampfer{at}obst.ucl.ac.be .
Bcl-2 mRNA expression was detected in rat blastocysts by in situ
hybridization. The distribution of mRNA expression was rather heterogenous,
with 
2% of high-expressing cells. In vitro exposure to 28 mmol/l
D-glucose for 24 h resulted in a significant increase in the proportion of
these cells compared with control embryos in either 6 mmol/l D-glucose or 28
mmol/l D+L-glucose. Heterogeneity in the expression of Bcl-2 was also observed
at the protein level by immunocytochemistry. Exposure to 28 mmol/l D-glucose
significantly increased the incidence of chromatin degradation (karyolysis)
and nuclear fragmentation (karyorhexis), two nuclear markers of apoptosis in
rat blastocysts. When two different antisense oligodeoxynucleotides designed
to block Bcl-2 expression were added to 28 mmol/l D-glucose, the incidence of
karyolysis (but not karyorhexis) was increased compared with embryos in 28
mmol/l D-glucose alone. These data suggest that Bcl-2 is involved in the
protective response against the induction of karyolysis in blastocysts on
their exposure to high concentrations of D-glucose in vitro, whereas
karyorhexis appears to result from the activation of an intracellular pathway
that is independent of Bcl-2.
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