| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1996: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Research Abstract |
The chromosomes of mouse embryos cultured under diabetic conditions were analyzed to clarify the causal mechanism of the congenital anomalies in the offspring of diabetic mothers. Mouse embryos used in this study were fertilized eggs, blastocysts and cells of day 8 embryos of gestation (vaginal plu=day 0). The embryos were cultured in CMRL 1066 (with 20% fetal calf serum) containing 300 mg/dl D (+) -glucose, 300 mg/dl D (+) -glucose plus 32 mM DL-beta-hydroxybutyric acid (DL-beta-OHB) or 32 mM DL-beta-OHB (keton body) alone for 2-5 days. 1) 1 cell culture (fertilized eggs) : Total of abnormal cells were 31.6% in the group exposed to glucose plus keton body, 30.6% in glucose and 20.5% in the control. There was no significant difference. High incidence in the control seems to be considerably influenced by culture. 2) Bastocyst culture : 14 Of 17 embryos (82.4%) had chromosomal anomalies in glucose and keton body, and 7 of 14 embryos (50.0%) in the glucose alone. Embryos of the two groups
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showed higher incidence of chromosomal anomalies than that of the control (4 of 15 embryos, 26.6%, P<0.01), and influence of glucose and keton body was suggested. 3) Cell culture (D8 embryo) : Numerical anomalies of chromosomes was 24.9% (P=.0.109, t-test) in glucose, 19.7% in keton body, 28.7% (P<0.01) in glucose plus keton body, whereas 18.7% in the control. NOR associations (14.7%, P<0.01) in glucose plus keton body increased in comparison with 2.6% in the control, which is compatible with the increase in aneuploidy upon addition of glucose plus keton body. Structural anomalies was 15.8% (P<0.01) in glucose, 18.5% (P<0.01) in keton body, 13.5% (P<0.01) in glucose plus keton body, whereas 2.0% in the control. Influence of glucose and keton body was suggested.
Histological cross-section of exencephalon from NOD-DM and STZ-diabetic mice (D12) was studied to find the relation between malformed tissue and chromosomal anomalies. Mitotic figuresalong the iuminal border in neuroepithelium were about 12% in exencephalon and normal, there was no significant difference. The relation between malformed tissue and chromosomal anomalies was not able to be found directly. Less
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