硬骨鱼STAT3剪接异构体在免疫系统中的表达调控与功能研究

项目名称： 硬骨鱼STAT3剪接异构体在免疫系统中的表达调控与功能研究

项目编号： No.31471181

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 汪新艳

作者单位： 电子科技大学

项目金额： 80万元

中文摘要： 选择性剪接是真核生物中的一种基本且重要的基因表达调节机制。我们在前期工作中发现草鱼、斑马鱼和金鱼的转录因子STAT3存在四种类似的选择性剪接体，进一步研究显示免疫刺激可诱导草鱼免疫细胞或组织中的STAT3剪接体的表达，且此诱导具有细胞因子选择性，提示鱼类STAT3信号通路具有高度的多样性和复杂性。本研究主要以草鱼为模型，针对STAT3剪接体的表达特征、转录因子特性和免疫学功能，采用qPCR、免疫细胞与组织化学染色、融合表达、ChIP-seq、蛋白质免疫印迹和免疫沉淀等方法，研究STAT3剪接体在草鱼免疫系统中产生和激活的规律及调控机制，阐明它们的转录活性差异及相互作用，明确它们在免疫调节中的功能地位。该研究不仅丰富鱼类转录因子选择性剪接和基于STAT3信号精细调控鱼类免疫反应的知识，而且提供STAT3的信号传导机制在脊椎动物中的进化信息。

中文关键词： 硬骨鱼；信号转导和转录活化因子3；选择性剪接；表达调控；功能地位

英文摘要： Alternative splicing is a fundamental and important regulatory mechanism in eukaryotes. We isolated four splicing variants of signal transducers and activators of transcription 3 (STAT3) from several teleosts including zebrafish, goldfish and grass carp. Using grass carp as the model, we found that immune stimuli (LPS, PHA and bacterial infection) could induce the expression of these STAT3 variants and this induction showed a cytokine specificity, indicating that the STAT3 signaling in fish immune system is highly diversified and complicated. In the present study, the expression patterns, transcriptional activities and immune functions of these four STAT3 variants will be investigated in grass carp, in particular elucidating the mechanisms of their generation and activation.Moreover,their transcriptional activity and interaction, and their roles in fish immunoregulation will be studied by using a serial of cell and molecular biological techniques including qPCR, cell and immunohistochemical staining, fusion protein expression, ChIP-seq, Western Blotting and Co-IP. This study will not only extend the knowledge about alternative splicing of fish transcription factor and fine regulatory mechanism of STAT3 signaling in fish immune response, but also provide the evolutional information of STAT3 signal transduction in vertebrates.

英文关键词： Teleost;STAT3;Alternative splicing;Expression regulation;Functional role