Heilig Charles W, Saunders Thomas, Brosius Frank C, Moley Kelle, Heilig Kathleen, Baggs Raymond, Guo LiRong, Conner David
Johns Hopkins University School of Medicine, 720 Rutland Avenue, Baltimore, MD 21205, USA.
Proc Natl Acad Sci U S A. 2003 Dec 23;100(26):15613-8. doi: 10.1073/pnas.2536196100. Epub 2003 Dec 12.
The hyperglycemia of maternal diabetes suppresses the glucose transporter-1 (GLUT1) facilitative glucose transporter 49-66% in preimplantation embryos. Glucose uptake is reduced and apoptosis is activated. We hypothesized that the reduction of embryonic GLUT1 may play a key role in the malformations of diabetic embryopathy. Therefore, we produced GLUT1-deficient transgenic mice [i.e., antisense-GLUT1 (GT1AS)] to determine whether GLUT1 deficiency alone could reproduce the growth defects. Early cell division of fertilized mouse eggs injected with GT1AS was markedly impaired, P < 0.001 vs. controls. Two populations of preimplantation embryos obtained from GT1AS x GT1AS heterozygote matings exhibited reduction of the 2-deoxyglucose uptake rate: one by 50% (presumed heterozygotes, P < 0.001 vs. control) and the other by 95% (presumed homozygotes, P < 0.001 vs. heterozygotes). Embryonic GLUT1 deficiency in the range reported with maternal diabetes was associated with growth retardation and developmental malformations similar to those described in diabetes-exposed embryos: intrauterine growth retardation (31.1%), caudal regression (9.8%), anencephaly with absence of the head (6.6%), microphthalmia (4.9%), and micrognathia (1.6%). Reduced body weight (small embryos, <70% of the nontransgenic body weight) was accompanied by other malformations and a 56% reduction of GLUT1 protein, P < 0.001 vs. nonsmall embryos (body weight >or=70% normal). The heart, brain, and kidneys of embryonic day 18.5 GT1AS embryos exhibited 24-51% reductions of GLUT1 protein. The homozygous GT1AS genotype was lethal during gestation. Reduced embryonic GLUT1 was associated with the appearance of apoptosis. Therefore, GLUT1 deficiency may play a role in producing embryonic malformations resulting from the hyperglycemia of maternal diabetes. Late gestational macrosomia was absent, apparently requiring a different mechanism.
母体糖尿病导致的高血糖会使植入前胚胎中的葡萄糖转运蛋白1(GLUT1)促进性葡萄糖转运减少49 - 66%。葡萄糖摄取减少,细胞凋亡被激活。我们推测胚胎GLUT1的减少可能在糖尿病胚胎病的畸形中起关键作用。因此,我们制备了GLUT1缺陷型转基因小鼠[即反义GLUT1(GT1AS)],以确定仅GLUT1缺陷是否能重现生长缺陷。注射了GT1AS的受精小鼠卵的早期细胞分裂明显受损,与对照组相比P < 0.001。从GT1AS×GT1AS杂合子交配获得的两个植入前胚胎群体显示2 - 脱氧葡萄糖摄取率降低:一个降低50%(推测为杂合子,与对照组相比P < 0.001),另一个降低95%(推测为纯合子,与杂合子相比P < 0.001)。胚胎GLUT1缺乏程度在母体糖尿病报道的范围内与生长迟缓及发育畸形有关,这些畸形与糖尿病暴露胚胎中描述的相似:宫内生长迟缓(31.1%)、尾椎退化(9.8%)、无脑伴头部缺失(6.6%)、小眼畸形(4.9%)和小颌畸形(1.6%)。体重减轻(小胚胎,<非转基因体重的70%)伴有其他畸形,且GLUT1蛋白减少56%,与非小胚胎(体重≥正常体重的70%)相比P < 0.001。胚胎第18.5天的GT1AS胚胎的心脏、大脑和肾脏中GLUT1蛋白减少24 - 51%。纯合GT1AS基因型在妊娠期致死。胚胎GLUT1减少与细胞凋亡的出现有关。因此,GLUT1缺乏可能在母体糖尿病高血糖导致的胚胎畸形中起作用。晚期妊娠巨大儿不存在,显然需要不同的机制。