Maternal hyperglycemia alters glucose transport and utilization in mouse preimplantation embryos
Maternal hyperglycemia alters glucose transport and utilization in mouse preimplantation embryos
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DOI：
10.1152/ajpendo.1998.275.1.e38
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发表时间：
1998-07-01
影响因子：
5.1
通讯作者：
Mueckler, MM
中科院分区：
文献类型：
--
作者：
Moley, KH;Chi, MMY;Mueckler, MM
研究了正常小鼠和糖尿病小鼠的植入前胚胎的葡萄糖利用情况。通过使用超微荧光酶测定，在交配后 24、48、72 和 96 小时测量从对照小鼠和链脲佐菌素诱导的高血糖小鼠中回收的单个胚胎中的胚胎内游离葡萄糖。糖尿病患者的游离葡萄糖浓度在 48 小时和 96 小时显着下降，对应于双细胞和囊胚阶段（48 小时：糖尿病 0.23 +/- 0.09 对比对照 2.30 +/- 0.43 mmol/kg 湿重；P < 0.001；96 小时：糖尿病 0.31 +/- 0.29 对比对照 5.12 +/- 0.17 mmol/kg湿重量；P < 0.001。然后使用非放射性 2-脱氧葡萄糖摄取和微荧光酶测定比较运输，糖尿病小鼠胚胎的 2-脱氧葡萄糖摄取在 48 小时和 96 小时均显着降低（48 小时糖尿病，0.037 +/- 0.003；对照，0.091）。 +/- 0.021 mmol.kg 湿重(-1).10 分钟(-1)，P < 0.05；96 小时糖尿病，0.249 +/- 0.008；对照，0.389 +/- 0.007 mmol.kg 湿重(-1).10 分钟(-1)，P < 0.02)。与对照小鼠相比，糖尿病小鼠在 48 小时（P < 0.001）和 96 小时（P < 0.05）时 GLUT-1 mRNA 分别减少 44% 和 68%。 96小时时GLUT-2和GLUT-3 mRNA值分别降低63%和77%（P < 0.01，P < 0.01）。定量免疫荧光显微镜显示，糖尿病胚胎中 48 小时和 96 小时的 GLUT-1 蛋白分别减少了 49 +/- 6 % 和 66 +/- 4%，96 小时的 GLUT-2 和 -3 蛋白分别减少了 90 +/- 5 和 84 +/- 6%。这些发现表明，为了应对母体糖尿病状态，植入前小鼠胚胎的葡萄糖利用率下降，这与 mRNA 和蛋白质水平上葡萄糖转运的减少直接相关。
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 min(-1), P < 0.05; 96 h diabetic, 0.249 +/- 0.008; control, 0.389 +/- 0.007 mmol.kg wet wt(-1).10 min(-1), 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.