PDF(Pubmed)
Abstract：
Ciliopathies are human genetic disorders caused by abnormal formation and dysfunction of cellular cilia. Cilia are microtubule-based organelles that project into the extracellular space and transduce molecular and chemical signals from the extracellular environment or neighboring cells. Intraflagellar transport (IFT) proteins are required for the assembly and maintenance of cilia by transporting proteins along the axoneme which consists of complexes A and B. IFT46, a core IFT-B protein complex, is required for cilium formation and maintenance during vertebrate embryonic development. Here, we introduce transgenic zebrafish lines under the control of ciliated cell-specific IFT46 promoter to recapitulate human ciliopathy-like phenotypes. We generated a Tg(IFT46:GAL4-VP16) line to temporo-spatially control the expression of effectors including fluorescent reporters or nitroreductase based on the GAL4/UAS system, which expresses GAL4-VP16 chimeric transcription factors in most ciliated tissues during embryonic development. To analyze the function of IFT46-expressing ciliated cells during zebrafish development, we generated the Tg(IFT46:GAL4-VP16;UAS;nfsb-mCherry) line, a ciliated cell-specific injury model induced by nitroreductase (NTR)/metrodinazole (MTZ). Conditionally, controlled ablation of ciliated cells in transgenic animals exhibited ciliopathy-like phenotypes including cystic kidneys and pericardial and periorbital edema. Altogether, we established a zebrafish NTR/MTZ-mediated ciliated cell injury model that recapitulates ciliopathy-like phenotypes and may be a vertebrate animal model to further investigate the etiology and therapeutic approaches to human ciliopathies.
摘要：
纤毛病是由细胞纤毛的异常形成和功能障碍引起的人类遗传性疾病。纤毛是基于微管的细胞器,其投射到细胞外空间并从细胞外环境或邻近细胞转导分子和化学信号。通过沿着由复合物A和B组成的轴突运输蛋白质来组装和维持纤毛所需的步内运输（IFT）蛋白质。IFT46是一种核心IFT-B蛋白质复合物，是脊椎动物胚胎发育过程中纤毛形成和维持所必需的。这里,我们在纤毛细胞特异性IFT46启动子的控制下引入转基因斑马鱼系，以概括人类纤毛病样表型。我们产生了Tg（IFT46：GAL4-VP16）系，以基于GAL4/UAS系统在时间空间上控制包括荧光报道分子或硝基还原酶在内的效应子的表达，在胚胎发育过程中，在大多数纤毛组织中表达GAL4-VP16嵌合转录因子。分析表达IFT46的纤毛细胞在斑马鱼发育过程中的功能,我们产生了Tg(IFT46:GAL4-VP16;UAS;nfsb-mCherry)线，由硝基还原酶（NTR）/美替硝唑（MTZ）诱导的纤毛细胞特异性损伤模型。有条件地,转基因动物纤毛细胞的受控消融表现为纤毛病样表型，包括囊性肾和心包及眶周水肿。总之,我们建立了斑马鱼NTR/MTZ介导的纤毛细胞损伤模型,该模型概括了纤毛病样表型,可能是进一步研究人类纤毛病的病因和治疗方法的脊椎动物模型.
