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Departments of 1 Obstetrics and Gynecology and of 2 Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110
Glucose
utilization was studied in preimplantation embryos from normal and
diabetic mice. With use of ultramicrofluorometric enzyme assays,
intraembryonic free glucose in single embryos recovered from
control and streptozotocin-induced hyperglycemic mice was measured at
24, 48, 72, and 96 h after mating. Free glucose concentrations dropped
significantly in diabetics at 48 and 96 h, corresponding to the
two-cell and blastocyst stages (48 h: diabetic 0.23 ± 0.09 vs.
control 2.30 ± 0.43 mmol/kg wet wt;
P < 0.001; 96 h: diabetic 0.31 ± 0.29 vs. control 5.12 ± 0.17 mmol/kg wet wt;
P < 0.001). Hexokinase activity was
not significantly different in the same groups. Transport was
then compared using nonradioactive 2-deoxyglucose uptake and
microfluorometric enzyme assays. The 2-deoxyglucose uptake was
significantly lower at both 48 and 96 h in embryos from
diabetic vs. control mice (48 h diabetic, 0.037 ± 0.003; control,
0.091 ± 0.021 mmol · kg wet
wt
1 · 10 min1,
P < 0.05; 96 h diabetic, 0.249 ± 0.008; control, 0.389 ± 0.007 mmol · kg wet
wt1 · 10 min1,
P < 0.02). When competitive
quantitative reverse transcription-polymerase chain reaction was used,
there was 44 and 68% reduction in the GLUT-1 mRNA at 48 h
(P < 0.001) and 96 h
(P < 0.05), respectively, in
diabetic vs. control mice. GLUT-2 and GLUT-3 mRNA values were decreased
63 and 77%, respectively (P < 0.01, P < 0.01) at 96 h. Quantitative
immunofluorescence microscopy demonstrated 49 ± 6 and 66 ± 4% less
GLUT-1 protein at 48 and 96 h and 90 ± 5 and 84 ± 6%
less GLUT-2 and -3 protein, respectively, at 96 h in diabetic embryos.
These findings suggest that, in response to a maternal diabetic state,
preimplantation mouse embryos experience a decrease in glucose
utilization directly related to a decrease in glucose transport at both
the mRNA and protein levels.
glucose toxicity; cleavage-stage embryos; glucose transport downregulation; maternal diabetes
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