转基因斑马鱼分析胰岛β-细胞发育情况(英文)  

ANALYSES OF β-CELL DEVELOPMENT IN TRANSGENIC ZEBRAFISH WITH A CONSTRUCT OF INSULIN PROMOTER AND GREEN FLUORESCENT PROTEIN

作　　者：夏铭[1] 潘雪[1] 李敏燕[1] 邓敏 张勇[3,4] 金怡[4] 陈漪[4] 王和生[1] 孔德明[1] 

机构地区：[1]贵阳中医学院,贵阳550002 [2]上海健康科学研究所,上海200025 [3]上海交通大学医学院,上海200025 [4]上海瑞金医院血液研究所,上海200025

出　　处：《水生生物学报》2009年第4期702-708,共7页Acta Hydrobiologica Sinica

基　　金：Founds of province(No.2006109);modernization of traditional Chinese medicine (No.20075007)

摘　　要：斑马鱼的个体小、高产和体外受精等特点使其已经迅速成为研究脊椎动物器官发育和人类疾病的模式生物之一。我们建立了一个转基因斑马鱼动物模型来研究胰岛β-细胞的发育。首先,构建了斑马鱼胰岛素(Insulin,INS)启动子与绿色荧光蛋白(GFP)组成的表达载体,命名为INS:GFP。其次,将质粒在斑马鱼1-细胞期注射到细胞质内。最后我们成功获得了生殖系稳定遗传胰岛素转基因斑马鱼,在成鱼和幼鱼期均可以通过GFP标记β-细胞。通过方便的荧光筛选,我们观察到胰岛在受精后18h开始形成,1—5d后由初始的脊索中线两侧向右迁移。从我们构建的胰岛素转基因斑马鱼,可以直观判断胰岛的发育情况,为研究胰岛的发育、损伤和再生提供了一个简便和直观的新型工具。With many advantageous features such as small size, high production and fertilization in vitro, zebrafish has become a kind of the model creature for the investigation of vertebrate development and human diseases. So we set up a transgenic zebrafish model to investigate the β-cell development. Firstly, we obtained an INS： GFP construction that contains the zebrafish insulin （INS） promoter and green fluorescent protein （GFP）. Secondly, we injected the construction into the cytoplasm of one-cell-stage embryos. Finally, we gained germline INS transgenic zebrafish that displayed highly specific β-cell expression of GFP in both larvae and adult. By following GFP expression, pancreas formation was detected at the 18h post-fertilization in the earliest time points between the second bilateral somites, a group of GFP-positive cells located from ventral to the notochord. From day 1 to 5 post-fertilization, the number of insulin/GFP expressing cells migrated and formed a right organ. Thus, our works demonstrated that the transgenic line provided a convenient and direct experimental tool in analyzing endocrine pancreas development, injury and recovery.

关 键 词：斑马鱼 Β-细胞 绿色荧光蛋白 胰岛素 

分 类 号：Q343[生物学—遗传学]