GAS41基因在斑马鱼胚胎发育中的时空表达谱

Temporal and Spatial Expression Profiling of GAS41 Gene in Zebrafish Embryonic Development

为了深入研究GAS41在胚胎发育中的作用机理,应用生物信息学软件分析了斑马鱼GAS41基因的序列特征;采用RT-PCR、整体原位杂交和整体免疫组织化学方法检测其在斑马鱼胚胎发育中的时空表达谱.结果显示斑马鱼GAS41蛋白与人类、小鼠、大鼠同源蛋白都有91.2%的同源性,具有高度的保守性.RT-PCR结果表明GAS41在检测的成体组织中均有表达,表达丰度相当.而GAS41在胚胎受精后一直持续表达,其中受精4 h后表达量升高.整体原位杂交和免疫化学结果显示GAS41特异性表达于胚胎头部,尤其是中脑、间脑(眼部)、小脑和后脑表达明显.这些都提示GAS41是一个高度保守基因,在斑马鱼中枢神经系统发育过程中可能起重要作用,同时也为GAS41在胚胎发育调控中的作用机制提供实验基础.

To further research the function and mechanism of GAS41 in embryonic development,the characteristic of zebrafish GAS41 gene sequence was analyzed by bioinformatics softwares,and the temporal and spatial expression profiling of zebrafish GAS41 in the embryonic development was detected by RT-PCR、whole-mount in situ hybridization and whole-mount immunostaining.The results showed zebrafish GAS41 protein,with highly conservative features,shares a 91.2% sequence identity to homologues of human,mouse and rat GAS41.RT-PCR revealed GAS41 expression was detected in all of adult tissues with the same abundance,and GAS41 continued to express after fertilization,its expression was increased in 4 hours post fertilization.The whole-mount in situ hybridization and whole-mount immunostaining showed GAS41 specifically expressed in the embryonic head region,especially in the midbrain,interbrain(eyes),cerebellum and hindbrain.These data suggest that GAS41 is a highly conserved gene and might play an important role in zebrafish central nerve system development,which also provides a solid experimental foundation for the research of the regulatory mechanism of GAS41 in embryonic development.