Cederberg J, Picard J J, Eriksson U J
Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden.
Diabetologia. 2003 Sep;46(9):1245-51. doi: 10.1007/s00125-003-1100-1. Epub 2003 Jun 27.
AIMS/HYPOTHESIS: Maternal diabetes mellitus increases the risk for fetal malformations. Several of these malformations are found in organs and tissues derived from the neural crest. Previous studies have shown changes in fetal organs of neural crest origin in experimental diabetes and changes in migration of neural crest cells exposed to high glucose in vitro.
We used whole-mount neurofilament staining of embryos from normal and diabetic mothers to investigate the development of cranial nerve ganglia. Neural tube explants were cultured in 10 and 40 mmol/l glucose and cell death and caspase activity was measured with flow cytometry.
The development of cranial ganglia V, VII, VIII, IX and X was impaired in day 10-11 embryos of diabetic rats. There was also a higher rate of cell death of neural crest derived cells cultured in 40 mmol/l glucose for 20 h (35% compared to 12% in 10 mmol/l). However, exposure of cells to 40 mmol/l glucose in culture did not increase the activation of the cell death effector proteins-caspases-measured as cellular binding of the activated caspase marker VAD-FMK. This suggests that the cell death is not caused by caspase-dependent apoptosis or that the caspases are activated at an earlier stage.
CONCLUSION/INTERPRETATION: The development of neural crest-derived structures is disturbed already at the organogenic period in embryos of diabetic rats and this deteriorated development could be due to high-glucose induced increase in cell death of neural crest derived cells.
目的/假设：母体糖尿病会增加胎儿畸形的风险。其中一些畸形出现在源自神经嵴的器官和组织中。先前的研究表明，实验性糖尿病中胎儿神经嵴来源器官发生了变化，体外暴露于高糖环境下的神经嵴细胞迁移也发生了变化。
我们对正常和糖尿病母亲的胚胎进行全胚胎神经丝染色，以研究颅神经节的发育。将神经管外植体培养于10和40 mmol/L葡萄糖中，通过流式细胞术检测细胞死亡和半胱天冬酶活性。
糖尿病大鼠第10 - 11天胚胎的V、VII、VIII、IX和X颅神经节发育受损。在40 mmol/L葡萄糖中培养20小时的神经嵴来源细胞的细胞死亡率也更高（35%，而10 mmol/L时为12%）。然而，在培养中将细胞暴露于4 mmol/L葡萄糖并未增加细胞死亡效应蛋白半胱天冬酶的激活，以活化的半胱天冬酶标记物VAD - FMK的细胞结合来衡量。这表明细胞死亡不是由半胱天冬酶依赖性凋亡引起的，或者半胱天冬酶在更早阶段被激活。
结论/解读：糖尿病大鼠胚胎在器官发生期神经嵴来源结构的发育就已受到干扰，这种发育恶化可能是由于高糖诱导神经嵴来源细胞的细胞死亡增加所致。
