Genetics of FAS in Mouse Component

产前大量酒精暴露可导致胎儿酒精综合征（FAS），其特征是生长 缺陷、面部畸形、脑损伤和神经行为功能障碍。这些异常情况 广泛地跨越现在被认为反映胎儿酒精谱系障碍（FASD）的范围。FASD变化 与怀孕期间饮酒模式的个体差异以及遗传和生物因素有关。 影响脆弱性的因素。关于FASD发病机制的分子机制知之甚少， 或关于导致FASD表现多样的遗传差异。我们的长期目标是 为了确定酒精对基因表达的影响，这种影响可能与大脑发育异常有关， 并鉴定影响对这些发病机制的易感性的遗传因素。我们已经使用 胚胎培养物，以使C56 BL/6（B6）和DBA/2（D2）近交系小鼠在早期神经胚形成期间暴露于酒精。 44小时后，B6和D2胚胎表现出不同的发育延迟和结构缺陷模式 暴露于高浓度酒精（300-400 mg/dl）的时间。在B6胚胎中，DMA微阵列 表达谱分析，然后是差异表达基因的假设驱动的生物信息学分析， 揭示了在调节特定功能的基因组中基因的过度表达。显著下调 许多对神经特化、神经分型和神经模式化至关重要的基因是显而易见的， 确认了几个。我们现在的目标是生成更详细的时间和空间分析 破坏基因组及其调节因子的表达，以确定与畸形的相关性， 这两个近交系的结果不同。目的1将鉴定B6和D2中的基因表达变化 胚胎后，无论是4或20小时的酒精暴露，使用DNA微阵列和假设驱动 生物信息学分析，以确定特定功能途径中基因组的影响。模式 将形态异常与用选择的原位杂交观察到的变化的定位进行比较。 基因.目的2将评估所观察到的选定小鼠基因下调与脊椎动物的相关性。 神经发育，通过使用新型斑马鱼吗啉敲除技术进行快速筛选 of losses损失of functional功能effects效果of orthopathy正交gene基因.目标3将使用信息学来确定酒精是否 影响可能是由于对特定转录因子（TF）功能的作用，通过鉴定预测的TF， 从TF结合位点常见的酒精影响的基因组在小鼠模型。这些研究将 提供了产前酒精对胚胎大脑发育的分子调节作用的见解，沿着 遗传因素影响与FAS相关的损伤的类型或程度。这些知识将 添加重要信息，用于识别、干预或治疗有FASD风险的妊娠。

Heavy prenatal alcohol exposure can cause fetal alcohol syndrome (FAS) characterized by growth deficits, facial dysmorphology, brain damage, and neurobehavioral dysfunction. These abnormalities vary widely across a range now considered to reflect a fetal alcohol spectrum disorder (FASD). Variation in FASD is linked to individual differences in patterns of drinking during pregnancy together with genetic and biological factors influencing vulnerability. Little is understood about the molecular mechanisms of FASD pathogenesis, or about genetic differences that contribute to the diverse FASD presentations. Our long-term objectives are to identify alcohol effects on gene expression that can link mechanistically to abnormal brain development, and to identify genetic factors that influence susceptibility to those pathogenesis mechanisms. We have used embryonic cultures to expose C56BL/6 (B6) and DBA/2 (D2) inbred mice to alcohol during early neurulation. The B6 and D2 embryos showed different patterns of developmental delays and structural defects after 44 hours of exposure to high alcohol concentrations (300-400 mg/dl). In the B6 embryos, DMA microarray expression profiling, followed by hypothesis-driven bioinformatics analysis of differentially expressed genes, revealed over-representation of genes in gene sets regulating specific functions. Prominent down regulation of many genes critical for neural specification, neural typing, and neural patterning was evident, and confirmed for several. Our goal now is to generate more detailed temporal and spatial analyses of the disrupted expression of gene cohorts and their regulators, to identify correlations with dysmorphology and differential outcomes in the two inbred strains. Aim 1 will identify gene expression changes in B6 and D2 embryos after either 4 or 20 hrs of alcohol exposure, using DNA microarray and hypothesis-driven bioinformatics analysis to identify effects in gene sets in specific functional pathways. Patterns of dysmorphology will be compared with localization of changes seen with in situ hybridization of selected genes. Aim 2 will assess the relevance of observed down regulation of selected mouse genes to vertebrate neural development, by using novel zebrafish morpholino knockdown technology to carry out rapid screening of loss-of-function effects of orthologous genes. Aim 3 will use informatics to determine whether alcohol effects may be due to actions on functions of specific transcription factors (TF), by identifying TFs predicted from TF binding sites common to alcohol-affected gene cohorts in the mouse models. These studies will provide insights into prenatal alcohol effects on molecular regulators of embryonic brain development, along with genetic factors that influence the type or extent of damage associated with FAS. This knowledge will add important information for the identification, intervention, or treatment in pregnancies at-risk for FASD.