PFOS致斑马鱼游囊发育畸形信号通路的研究

项目名称： PFOS致斑马鱼游囊发育畸形信号通路的研究

项目编号： No.21277104

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 环境科学、安全科学

项目作者： 杨东仁

作者单位： 温州医科大学

项目金额： 76万元

中文摘要： PFOS是一种新型持久性有机污染物，也是迄今为止人体内检测到的含量最高的外源性化合物。申请者近期发现PFOS对斑马鱼胚胎游囊发育有典型的畸形效应，且与全基因组微阵列分析结果相吻合。本项目以IPA构建的PFOS响应的游囊畸形效应信号通道（59个基因）为研究对象，用RT-PCR重新精确定量，再用原位杂交（ISH）对具显著变化的基因定位研究，从中筛选出6-8个PFOS响应的且在游囊表达的备选基因。第二部分，用显微注射MO或mRNA人为干扰基因表达的方法，结合斑马鱼转基因品系、共聚焦显微观察、ISH、RT-PCR、western blot、免疫组织化学、细胞凋亡、显微切片等实验手段，研究备选基因在基因和蛋白层面上的功能，最终筛选出1-2个PFOS响应的且在游囊发育过程中具重要生命功能的靶基因。研究成果对PFOS响应的新型分子标志物的筛选及未来构建响应PFOS的实时监测报告基因鱼等均具有重要的意义。

中文关键词： PFOS；斑马鱼；游囊发育毒性；信号通路；靶基因筛选

英文摘要： Perfluorooctanesulphonic acid (PFOS), a new kind of persistent organic pollutants, is also one of the exogenous compounds that has been detected with high levels in human body . Recently we found PFOS induced embryonic zebrafish swim bladder deformity and that was consistent with the whole genomic microarray analysis results. Using IPA software, we constructed a swim bladder deformity signaling pathway perturbed by PFOS. Based on this pathway, this project will use RT-PCR method to precisely quantify the gene expression, and then use in situ hybridization (ISH) approach to localize the significant changed genes in embryonic zebrafish. Finally we expect to screen out PFOS responsive genes and their expression leves in the swim bladder. In the second part, we will use interference gene expression approaches such as microinjection MO (down regulate gene expression) or mRNA (up regulate gene expression) to study the function of these genes as well as use western blot or immunohistochemistry methods to study their protein expression. In addition, we will also take advantage of some transgenic zebrafish models, to identify the target genes that are mediating PFOS induced swim bladder deformities. Our results can be used to screen PFOS exposure responsive molecular biomarkers and for construction of a trangenic report

英文关键词： PFOS；zebrafish；developmental toxicity of swim bladder；signal pathway；target gene screening