Savage Daniel D, Rosenberg Martina J, Coquet Laurent, Porch Morgan W, Allen Nyika A, Roux Christian, Aligny Caroline, Jouenne Thierry, Gonzalez Bruno J
Department of Neurosciences, School of Medicine, University of New Mexico, Albuquerque, NM, United States.
UMR 6270, CNRS, Normandie University, UNIROUEN, Proteomic Facility PISSARO, Institute for Research and Innovation in Biomedicine, Rouen, France.
Front Neurosci. 2020 Jun 3;14:519. doi: 10.3389/fnins.2020.00519. eCollection 2020.
Jegou et al. (2012) have reported prenatal alcohol exposure (PAE)-induced reductions of angiogenesis-related proteins in mouse placenta. These effects were associated with striking alterations in microvascular development in neonatal cerebral cortex. Here, we employed a rat model of moderate PAE to search for additional proteins whose placental and fetal cortical expression is altered by PAE, along with a subsequent examination of fetal cerebral cortical alterations associated with altered protein expression. Long-Evans rat dams voluntarily consumed either a 0 or 5% ethanol solution 4 h each day throughout gestation. Daily ethanol consumption, which resulted in a mean peak maternal serum ethanol concentration of 60.8 mg/dL, did not affect maternal weight gain, litter size, or placental or fetal body weight. On gestational day 20, rat placental: fetal units were removed by Caesarian section. Placental protein expression, analyzed by 2D-PAGE - tandem mass spectroscopy, identified a total of 1,117 protein spots, 20 of which were significantly altered by PAE. To date, 14 of these PAE-altered proteins have been identified. Western blotting confirmed the alterations of two of these placental proteins, namely, annexin-A4 (ANX-A4) and cerebral cavernous malformation protein 3 (CCM-3). Specifically, PAE elevated ANX-A4 and decreased CCM-3 in placenta. Subsequently, these two proteins were measured in fetal cerebral cortex, along with radiohistochemical studies of VEGF binding and histofluorescence studies of microvascular density in fetal cerebral cortex. PAE elevated ANX-A4 and decreased CCM-3 in fetal cerebral cortex, in a pattern similar to the alterations observed in placenta. Further, both VEGF receptor binding and microvascular density and orientation, measures that are sensitive to reduced CCM-3 expression in developing brain, were significantly reduced in the ventricular zone of fetal cerebral cortex. These results suggest that the expression angiogenesis-related proteins in placenta might serve as a biomarker of ethanol-induced alterations in microvascular development in fetal brain.
热古等人（2012年）报告称，产前酒精暴露（PAE）会导致小鼠胎盘中血管生成相关蛋白减少。这些影响与新生小鼠大脑皮质微血管发育的显著改变有关。在此，我们采用中度PAE大鼠模型，寻找胎盘和胎儿皮质表达因PAE而改变的其他蛋白，并随后检查与蛋白表达改变相关的胎儿大脑皮质改变。长-伊斯特大鼠母鼠在整个妊娠期每天自愿饮用4小时的0%或5%乙醇溶液。每日乙醇摄入量导致母体血清乙醇平均峰值浓度为60.8毫克/分升，这并未影响母体体重增加、窝仔数或胎盘及胎儿体重。在妊娠第20天，通过剖腹产取出大鼠胎盘-胎儿单元。通过二维聚丙烯酰胺凝胶电泳-串联质谱分析胎盘蛋白表达，共鉴定出1117个蛋白点，其中20个因PAE而发生显著改变。迄今为止，已鉴定出其中14种因PAE改变的蛋白。蛋白质印迹法证实了其中两种胎盘蛋白即膜联蛋白A4（ANX-A4）和脑海绵状血管畸形蛋白3（CCM-3）的改变。具体而言，PAE使胎盘中的ANX-A4升高，CCM-3降低。随后，在胎儿大脑皮质中检测了这两种蛋白，并对胎儿大脑皮质中血管内皮生长因子（VEGF）结合进行了放射组织化学研究以及对微血管密度进行了组织荧光研究。PAE使胎儿大脑皮质中的ANX-A4升高，CCM-3降低，其模式与在胎盘中观察到的改变相似。此外，在胎儿大脑皮质的脑室区，VEGF受体结合以及微血管密度和方向（这些指标对发育中的大脑中CCM-3表达降低敏感）均显著降低。这些结果表明，胎盘中血管生成相关蛋白的表达可能作为乙醇诱导胎儿大脑微血管发育改变的生物标志物。
