Department of Obstetrics and Gynecology, Washington University School of Medicine, 425 South Euclid Avenue, Campus Box 8064, St. Louis, Missouri 63110, USA.
Endocrinology. 2012 Apr;153(4):1984-9. doi: 10.1210/en.2011-1815. Epub 2012 Jan 31.
Maternal diabetes has been demonstrated to adversely affect preimplantation embryo development and pregnancy outcomes. Emerging data suggest that these effects are associated with compromised oocyte quality. However, direct evidence of a pathway by which maternal diabetes exerts its effects on the oocyte is still lacking. Cumulus cells are metabolically coupled to oocytes, and bidirectional communication between them is essential for the development and functions of both compartments. The primary focus of this work was to evaluate the connection between glucose uptake in cumulus cells and oocyte quality in diabetic mice. This experiment has been difficult, because cumulus cells need to be separated from oocytes and labeled with isotope in the process of measuring glucose uptake. Here, we report a method for live imaging glucose transport in single cumulus-oocyte complexes using a fluorescent glucose analog (6-(N-(7-nitrobenz-2-oxa-1,3-diazol- 4-yl)amino)-6-deoxyglucose). By tracking the ATP content and spindle/chromosome status in individual oocytes surrounded by cumulus cells with differing glucose uptake activity, we reveal that compromised oocyte quality in diabetic mice is linked to decreased glucose uptake in cumulus cells.
母体糖尿病已被证明会对胚胎着床前的胚胎发育和妊娠结局产生不利影响。新出现的数据表明,这些影响与卵母细胞质量受损有关。然而,母体糖尿病对卵母细胞产生影响的途径的直接证据仍然缺乏。卵丘细胞与卵母细胞代谢偶联,它们之间的双向通讯对两者的发育和功能都是至关重要的。这项工作的主要重点是评估糖尿病小鼠卵丘细胞中的葡萄糖摄取与卵母细胞质量之间的联系。由于在测量葡萄糖摄取的过程中需要将卵丘细胞从卵母细胞中分离出来并用同位素进行标记,因此这个实验具有一定的难度。在这里,我们报告了一种使用荧光葡萄糖类似物(6-(N-(7-硝基苯并-2-氧代-1,3-二唑-4-基)氨基)-6-脱氧葡萄糖)对单个卵丘-卵母细胞复合物中的葡萄糖转运进行实时成像的方法。通过跟踪具有不同葡萄糖摄取活性的卵丘细胞包围的单个卵母细胞中的 ATP 含量和纺锤体/染色体状态,我们揭示了糖尿病小鼠中卵母细胞质量受损与卵丘细胞中葡萄糖摄取减少有关。