Mypt1在斑马鱼前肾发育中功能与机制的研究

项目名称： Mypt1在斑马鱼前肾发育中功能与机制的研究

项目编号： No.31201085

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 发育生物学与生殖生物学

项目作者： 阮华

作者单位： 西南大学

项目金额： 24万元

中文摘要： 许多肾脏疾病是肾脏发育调控基因突变所导致的，研究肾脏发育对于治疗肾脏疾病意义重大。肾脏发育自中胚层，斑马鱼前肾是哺乳动物后肾的对应物，其发育的分子机制是保守的，是研究肾脏发育的理想模型。我们发现斑马鱼mypt1突变体早期前肾的特化、分化及各节段定位正常，30hpf时突变体前肾进曲小管PCT前段部分缺失,颈部结构也未处于预定位置。Mypt1调控中胚层细胞的迁移，突变体的表型很可能是前肾细胞迁移异常导致的。本课题运用细胞实时追踪这一斑马鱼模式生物的优势技术，设计以下实验研究Mypt1在斑马鱼前肾发育中的功能与机制：1）mypt1突变体前肾发育缺陷分析；2）mypt1突变体表型自主性的研究；3）建立PCT细胞和前肾颈部特异性启动子驱动KEADE的转基因鱼，研究mypt1突变体中PCT细胞和前肾颈部的迁移与定位；4）检测与中胚层细胞迁移有关的因素，发掘突变体表型的内在分子机制研究。

中文关键词： 斑马鱼；前肾；Mypt1；胰腺；

英文摘要： Many kidney diseases are caused by mutations in kidney development regulatory genes; studies of kidney development will promote clinical treatment of kidney diseases. All of vertebrate kidneys are derived from mesoderm, the zebrafish pronephros, the functional counterpart of metanephros in mammalians, is an ideal model to study kidney development due to conserved molecular mechanisms across vertebrates. We found that the loss-of-function mutation in zebrafish mypt1 (myosin phosphatase target subunit 1)did not disrupt specification, segmental differentiation and positioning of pronephros at 24hpf, while at 30hpf the proximal convoluted segment (PCT) in the mutant was properly differentiated, with a loss of anterior part, and the neck of mutant pronephros was located more posteriorly than the wild type. Since Mypt1 regulates mesoderm cell migration, we speculated that this mutant phenotype was caused by the defective cell migration of renal mesoderm. To address the mechanism underlying this phenotype,we propose following experiments: Firstly, detailed characterization of pronephric phenotype in mypt1 mutant. Secondly,to examine autonomy of phenotype caused by the mutation. Thirdly, to generate pronephric PCT and neck tissue specific promoters driven KEADE transgenic lines to study migration and positioning of ma

英文关键词： zebrafish；pronephros；Mypt1；pancreas；