Fetal alcohol syndrome (FAS) in C57BL/6 mice detected through proteomics screening of the amniotic fluid
Fetal alcohol syndrome (FAS) in C57BL/6 mice detected through proteomics screening of the amniotic fluid
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DOI：
10.1002/bdra.20440
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发表时间：
2008-04-01
影响因子：
--
通讯作者：
Knudsen, Thomas B.
中科院分区：
文献类型：
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作者：
Datta, Susmita;Turner, Delano;Knudsen, Thomas B.
背景技术背景：胎儿酒精综合征（FAS）是胎儿酒精谱系障碍的严重后果，与颅面缺陷、智力迟钝和发育迟缓有关。先前在C57 BL/6 J和C57 BL/6 N小鼠中的研究提供了证据，即酒精诱导的发病机制遵循胚胎头褶中特定分子途径内基因表达的早期变化。而前者（B6 J）妊娠进行高风险的畸形后，母体暴露于2.9 g/kg的酒精（两次注射间隔4.0小时，在妊娠第8天），后者（B6 N）妊娠进行低风险的畸形。本研究使用这种小鼠模型来筛选羊水中可能区分FAS阳性和FAS阴性妊娠的生物标志物。方法：B6 J和B6 N窝在妊娠第8天用酒精（暴露）或生理盐水（对照）处理。在第17天抽吸的羊水（每组n = 6个重复的窝）经受胰蛋白酶消化，以通过基质辅助激光解吸-飞行时间质谱法借助于去噪算法、统计检验和分类方法进行分析。结果：我们在蛋白质组学筛选中确定了几个峰，这些峰在暴露的B6 J窝仔中一致且特异性地降低。通过液相色谱串联质谱和多维蛋白质鉴定进行的初步表征将还原峰映射到甲胎蛋白（AFP）。通过受试者工作特征曲线下面积证实AFP缺乏作为FAS阳性窝仔生物标志物的预测强度。结论：这些在遗传易感小鼠中的发现支持了母体血清中的临床观察结果，该观察结果涉及产前酒精损伤后AFP水平的降低。
BACKGROUND: Fetal Alcohol Syndrome (FAS), a severe consequence of the Fetal Alcohol Spectrum Disorders, is associated with craniofacial defects, mental retardation, and stunted growth. Previous studies in C57BL/6J and C57BL/6N mice provide evidence that alcohol-induced pathogenesis follows early changes in gene expression within specific molecular pathways in the embryonic headfold. Whereas the former (B6J) pregnancies carry a high-risk for dysmorphogenesis following maternal exposure to 2.9 g/kg alcohol (two injections spaced 4.0 h apart on gestation day 8), the latter (B6N) pregnancies carry a low-risk for malformations. The present study used this murine model to screen amniotic fluid for biomarkers that could potentially discriminate between FAS-positive and FAS-negative pregnancies. METHODS: B6J and B6N litters were treated with alcohol (exposed) or saline (control) on day 8 of gestation. Amniotic fluid aspirated on day 17 (n = 6 replicate litters per group) was subjected to trypsin digestion for analysis by matrix-assisted laser desorption-time of flight mass spectrometry with the aid of denoising algorithms, statistical testing, and classification methods. RESULTS: We identified several peaks in the proteomics screen that were reduced consistently and specifically in exposed B6J litters. Preliminary characterization by liquid chromatography tandem mass spectrometry and multidimensional protein identification mapped the reduced peaks to alpha fetoprotein (AFP). The predictive strength of AFP deficiency as a biomarker for FAS-positive litters was confirmed by area under the receiver operating characteristic curve. CONCLUSIONS: These findings in genetically susceptible mice support clinical observations in maternal serum that implicate a decrease in AFP levels following prenatal alcohol damage.