Expression data from adult wild-type and zCdx1b transgenic zebrafish (3 months old). Danio rerio

Cdx2 has been suggested to play an important role in Barrett's esophagus (BE), or intestinal metaplasia (IM) in the esophagus. However, in vivo data have been lacking. The aim of the present study was to investigate whether transgenic overexpression of zCdx1b, the functional equivalent of mammalian Cdx2 in zebrafish, may lead to IM of squamous epithelium in zebrafish A transgenic zebrafish system was developed by expressing zCdx1b gene under the control of zebrafish keratin 5 promoter (zK5p). zCdx1b expression in the esophageal squamous epithelium of transgenic zebrafish was analyzed by in situ hybridization, immunohistochemical staining and RT-PCR. Gene expression in the esophageal squamous epithelium of wild-type and transgenic zebrafish was analyzed by Affymetrix microarray and confirmed by in situ hybridization. Overall design: The upper digestive tract tissue from 3 adult zebrafish (3 months old) was pooled as one sample. Three zCdx1b transgenic samples were used for microarray, and 2 wild type samples were used for control.

已有研究提示Cdx2在巴雷特食管（Barrett esophagus, BE）及食管肠上皮化生（intestinal metaplasia, IM）中发挥重要作用。然而，目前仍缺乏相关体内实验数据。
本研究旨在探究：在斑马鱼中功能等同于哺乳动物Cdx2的zCdx1b转基因过表达，是否会诱导斑马鱼鳞状上皮发生肠上皮化生。
本研究通过在斑马鱼角蛋白5启动子（zebrafish keratin 5 promoter, zK5p）的调控下表达zCdx1b基因，构建了转基因斑马鱼模型。通过原位杂交（in situ hybridization）、免疫组化染色（immunohistochemical staining）及逆转录PCR（RT-PCR），分析了转基因斑马鱼食管鳞状上皮中zCdx1b的表达水平。
采用Affymetrix基因芯片分析野生型与转基因斑马鱼食管鳞状上皮的基因表达谱，并通过原位杂交验证芯片检测结果。
实验设计：将3只3月龄成年斑马鱼的上消化道组织混合为一个样本；共设置3个zCdx1b转基因样本用于基因芯片检测，2个野生型样本作为对照。