Expression profiles of zebrafish transgenic line Tg(fabp10a:def)-I

We previously reported that Def (Digestive-organ expansion factor) was a pan-endodermal enriched factor that is essential for the growth of digestive organs in zebrafish using a def mutant line hi429 as model (Chen et al., 2005). To further elucidate Def function, we generated a Def over-expressed zebrafish line, namely Tg (fabp10a:def)-I, in which def expression was under the control of a liver-specific promoter fabp10a. We used microarrays to detail the global programme of gene expression in Tg (fabp10a:def)-I transgenic line compared with wild type zebrafish control, and identified distinct classes of differently regulated genes in Tg (fabp10a:def)-I line. Total RNA from the adult livers of three independent batches of 3-month-old Tg(fabp10a:def)-I and wild type (AB strain) male fish was extracted using TRIzol (Invitrogen), each batch with the liver samples from three fish mixed together. The samples were subjected to microarray hybridization and the data were generated for further analysis.

我们此前曾以def突变体品系hi429为模型，在斑马鱼中报道过Def（Digestive-organ expansion factor，消化器官扩增因子）是一种泛内胚层富集因子，对斑马鱼消化器官的生长至关重要（Chen等人，2005）。为进一步阐明Def的功能，我们构建了过表达Def的斑马鱼品系Tg(fabp10a:def)-I，该品系中def的表达受肝脏特异性启动子fabp10a调控。我们通过基因芯片（microarray）技术，对比分析了Tg(fabp10a:def)-I转基因斑马鱼与野生型对照斑马鱼的全基因表达谱，鉴定出该品系中存在多类差异调控的基因。我们使用TRIzol（Invitrogen）试剂盒，从3批独立的3月龄Tg(fabp10a:def)-I转基因雄鱼与野生型AB品系雄鱼的成体肝脏中提取总RNA，每批样本均混合3尾鱼的肝脏组织。将上述样本进行基因芯片杂交实验，最终获得可用于后续分析的实验数据。