Yang Penghua, Shen Wei-bin, Reece E Albert, Chen Xi, Yang Peixin
Department of Obstetrics, Gynecology & Reproductive Sciences, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Department of Obstetrics, Gynecology & Reproductive Sciences, University of Maryland School of Medicine, Baltimore, MD 21201, USA; Department of Biochemistry & Molecular Biology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
Biochem Biophys Res Commun. 2016 Apr 1;472(2):306-12. doi: 10.1016/j.bbrc.2016.02.117. Epub 2016 Mar 3.
Abnormal neurogenesis occurs during embryonic development in human diabetic pregnancies and in animal models of diabetic embryopathy. Our previous studies in a mouse model of diabetic embryopathy have implicated that high glucose of maternal diabetes delays neurogenesis in the developing neuroepithelium leading to neural tube defects. However, the underlying process in high glucose-impaired neurogenesis is uncharacterized. Neurogenesis from embryonic stem (ES) cells provides a valuable model for understanding the abnormal neural lineage development under high glucose conditions. ES cells are commonly generated and maintained in high glucose (approximately 25 mM glucose). Here, the mouse ES cell line, E14, was gradually adapted to and maintained in low glucose (5 mM), and became a glucose responsive E14 (GR-E14) line. High glucose induced the endoplasmic reticulum stress marker, CHOP, in GR-E14 cells. Under low glucose conditions, the GR-E14 cells retained their pluripotency and capability to differentiate into neural lineage cells. GR-E14 cell differentiation into neural stem cells (Sox1 and nestin positive cells) was inhibited by high glucose. Neuron (Tuj1 positive cells) and glia (GFAP positive cells) differentiation from GR-E14 cells was also suppressed by high glucose. In addition, high glucose delayed GR-E14 differentiation into neural crest cells by decreasing neural crest markers, paired box 3 (Pax3) and paired box 7 (Pax7). Thus, high glucose impairs ES cell differentiation into neural lineage cells. The low glucose adapted and high glucose responsive GR-E14 cell line is a useful in vitro model for assessing the adverse effect of high glucose on the development of the central nervous system.
异常神经发生在人类糖尿病妊娠的胚胎发育过程中以及糖尿病胚胎病的动物模型中出现。我们之前在糖尿病胚胎病小鼠模型中的研究表明，母体糖尿病的高血糖会延迟发育中的神经上皮中的神经发生，导致神经管缺陷。然而，高血糖损害神经发生的潜在过程尚不清楚。胚胎干细胞（ES细胞）的神经发生为理解高血糖条件下异常神经谱系发育提供了一个有价值的模型。ES细胞通常在高葡萄糖（约25 mM葡萄糖）中产生和维持。在这里，小鼠ES细胞系E14逐渐适应并维持在低葡萄糖（5 mM）中，并成为葡萄糖反应性E14（GR-E14）细胞系。高葡萄糖在GR-E14细胞中诱导内质网应激标志物CHOP。在低葡萄糖条件下，GR-E14细胞保持其多能性和分化为神经谱系细胞的能力。高葡萄糖抑制GR-E14细胞分化为神经干细胞（Sox1和巢蛋白阳性细胞）。GR-E14细胞向神经元（Tuj1阳性细胞）和神经胶质细胞（GFAP阳性细胞）的分化也受到高葡萄糖的抑制。此外，高葡萄糖通过降低神经嵴标志物配对盒3（Pax3）和配对盒7（Pax7）来延迟GR-E14向神经嵴细胞的分化。因此，高葡萄糖损害ES细胞向神经谱系细胞的分化。低葡萄糖适应且高葡萄糖反应性的GR-E14细胞系是评估高葡萄糖对中枢神经系统发育的不利影响的有用体外模型。
