MicroRNA靶向调控OSTF1在氟中毒鱼鳃渗透调节中的作用机制

项目名称： MicroRNA靶向调控OSTF1在氟中毒鱼鳃渗透调节中的作用机制

项目编号： No.31502141

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 农业科学

项目作者： 曹谨玲

作者单位： 山西农业大学

项目金额： 22万元

中文摘要： 氟是对生物危害严重的主要环境污染物。部分养殖水体含氟量高，对鱼类产生毒害效应。申请者前期研究发现，OSTF1阳性细胞主要位于鳃丝上皮细胞，且随着氟暴露浓度的增加，OSTF1蛋白表达增加，但氟诱导OSTF1表达的调控机制并不清楚。本项目拟在前期研究基础上，模拟水氟污染，以氟暴露鱼鳃及鳃细胞为材料，利用生物信息学及细胞培养、转染、基因沉默、荧光定量PCR、免疫印迹和免疫组化等细胞生物学和分子生物学等技术，预测靶向调控OSTF1的microRNAs并进行验证，对候选microRNAs和OSTF1的相关性进行分析，并研究microRNA mimic/inhibitor对氟中毒鱼鳃细胞渗透调节因子Na+-K+-ATPase、CFTR、OSTF1、AQP-3和NHE-1基因和蛋白表达的影响，探讨microRNAs靶向调控OSTF1在氟中毒鱼鳃渗透调节中的作用，从分子水平揭示氟对鱼鳃渗透调节影响的机理。

中文关键词： 水产动物；环境胁迫；抗逆；响应；基因表达调控

英文摘要： Fluoride is one of the main environmental pollutants that harms the health of organisms. High levels of fluoride in the fish ponds caused damages to fish. Our early results demonstrated that positive cells of OSTF1 were mainly located in epithelial cells of branchial filaments. With the increase of fluoride concentration, the protein expression of OSTF1 increased. However, it is unclear that the functional mechanisms of fluoride-induced expression of OSTF1 in the gills in common carp exposed to high fluoride. The proposed research intends to predict and verify microRNAs regulating OSTF1, analysis the correlation of candicate microRNAs and OSTF1, and study the effects of microRNA mimic/inhibitor on the osmotic regulation related factors in branchial cells, such as Na+/K+-ATPase, Osmotic regulation of transcription factors 1 (OSTF1), Cystic fibrosis transmembrane regulator (CFTR), Aquaporin 3(AQP-3), Sodium hydrogen exchanger1 (NHE-1) using bioinformatics, cell culture, transfection, gene silencing, real-time quantitative PCR, Western blotting and immunohistochemistry methods in fluoride-exposed gills or branchial cells based on their pollution situations in the water. Our study will clarify functional mechanism of microRNAs’ regulation to OSTF1 in osmotic adjustment of gills in common carp exposed to high fluoride. Our results could also reveal the mechanism of the effects of fluoride on osmotic regulation in fish from the molecular levels.

英文关键词： aquatic animals ;environmental stress;resilience;response;regulation of gene expression