Strain-dependent effects of alcohol on early mouse embryos

EXPERIMENT: Microarray expression profiles derived from the cranial neural folds (headfold) of neurulation-stage mouse embryos 3.0h after maternal alcohol exposure on 8 d.p.c. ANIMAL MODEL: Pregnancies from 2 substrains of C57BL/6 mice that differ in relative risk of Fetal Alcohol Syndrome (FAS): C57BL/6J (B6J) high-risk condition, and C57BL/6NCrl (B6N) low-risk condition. EXPOSURE: Dams dosed with ethanol (EtOH) by intraperitoneal injection at 2.9 g EtOH per kg body weight. Controls received vehicle (saline) alone. A third group of dams received EtOH + 4.0 mg/kg PK11195, a specific high-affinity ligand to the 18 KDa mitochondrial peripheral benzodiazepine receptor site. INTERVAL: High blood alcohol content must be sustained in dams for several hours to invoke FAS and is traditionally accomplished by a double injection of EtOH 4.0h apart. Since these embryos were harvested for genetic analysis 1h before dams would have gotten the second alcohol injection, the interval represents a snapshot of the critical response, prior to the second maternal injection that invokes greater risk. Counter-exposure to PK11195 significantly lowers the adverse response of B6J embryos to EtOH. PLATFORM: Two independent assays run. The first dataset (PE), run April 2002 – January 2003, used samples pooled from 2 litters (PE) and the platform was a two-channel MPS621 array (http://www.lifesciences.perkinelmer.com/). This platform has 4800 sequence-verified gene elements derived from over 50 different human cDNA libraries reflecting a variety of well-annotated cellular processes and disease pathways. The second dataset (AF), run between May 2005 – November 2005, used samples pooled from 1 litter and the platform was Affymetrix GeneChip® Mouse Genome 430A 2.0 Array (http://www.affymetrix.com/). The MG430A 2.0 platform has 45,102 oligonucleotide probe cells representing approximately 14,000 well-characterized genes in the draft mouse genome assembly. The corresponding GEO samples are GSM12218 - GSM12227 for the two-channel PE arrays, and GSM136067 - GSM136078 for the one-channel AF arrays. Keywords = fetal alcohol syndrome Keywords = alcohol-related birth defects Keywords = EtOH Keywords = PK11195 Keywords = embryo Keywords = strain differences Keywords: Ordered Assay of early mouse embryos during pathogenesis of Fetal Alcohol Syndrome (FAS). Replicate RNA samples were arrayed by one-channel oligonucleotide (Affymetrix) or two-channel cDNA (Perkin-Elmer) platforms.

实验：采集妊娠第8天（8 d.p.c.）神经胚形成期小鼠胚胎颅神经褶（头褶）的微阵列（Microarray）表达谱，样本取自母鼠酒精暴露3.0小时后。
动物模型：使用两种亚系的C57BL/6小鼠构建妊娠模型，二者胎儿酒精综合征（Fetal Alcohol Syndrome, FAS）的相对发病风险存在差异：其中C57BL/6J（B6J）为高风险亚系，C57BL/6NCrl（B6N）为低风险亚系。
暴露处理：孕鼠经腹腔注射给予乙醇（ethanol, EtOH），剂量为每千克体重2.9g乙醇。对照组孕鼠仅给予赋形剂（生理盐水）。第三组孕鼠给予乙醇联合4.0mg/kg PK11195——一种针对18kDa线粒体外周苯二氮䓬受体位点的特异性高亲和力配体。
采样间隔：为诱发FAS，需维持孕鼠血液高乙醇浓度数小时，传统方案采用两次间隔4.0小时的乙醇注射。本研究中，胚胎采集于孕鼠即将接受第二次乙醇注射前1小时，因此该采样间隔捕捉的是第二次高风险注射前的关键应激反应快照。PK11195干预可显著降低B6J小鼠胚胎对乙醇的不良应答反应。
检测平台：本研究包含两个独立检测数据集。第一组数据集（PE）于2002年4月至2003年1月完成，样本混合自2窝幼崽，采用双通道MPS621微阵列平台（http://www.lifesciences.perkinelmer.com/）。该平台包含4800个经序列验证的基因探针元件，源自50余个不同的人类cDNA文库，覆盖多种已注释的细胞过程与疾病通路。第二组数据集（AF）于2005年5月至2005年11月完成，样本混合自1窝幼崽，采用Affymetrix GeneChip® Mouse Genome 430A 2.0基因芯片平台（http://www.affymetrix.com/）。MG430A 2.0平台包含45102个寡核苷酸探针单元，对应小鼠基因组草图组装版本中约14000个已充分注释的基因。双通道PE阵列对应的GEO样本编号为GSM12218至GSM12227，单通道AF阵列对应的GEO样本编号为GSM136067至GSM136078。
关键词：胎儿酒精综合征、酒精相关出生缺陷、乙醇、PK11195、胚胎、品系差异。研究主题：胎儿酒精综合征发病进程中早期小鼠胚胎的有序检测分析。重复RNA样本分别通过单通道寡核苷酸（Affymetrix）或双通道cDNA（Perkin-Elmer）平台完成微阵列检测。