Morales Elvin E, Wingert Rebecca A
Department of Biological Sciences, Center for Stem Cells and Regenerative Medicine, Center for Zebrafish Research, University of Notre Dame, 100 Galvin Life Sciences, Notre Dame, IN, 46556, USA.
Results Probl Cell Differ. 2017;60:55-75. doi: 10.1007/978-3-319-51436-9_3.
Animal models have been an invaluable means to advance biomedical research as they provide experimental avenues for cellular and molecular investigations of disease pathology. The zebrafish (Danio rerio) is a good alternative to mammalian models that can be used to apply powerful genetic experimental methods normally used in invertebrates to answer questions about vertebrate development and disease. In the case of the kidney, the zebrafish has proven itself to be an applicable and versatile experimental system, mainly due to the simplicity of its pronephros, which contains two nephrons that possess conserved structural and physiological aspects with mammalian nephrons. Numerous genes that were not previously related to kidney conditions have now been linked to renal diseases by applying genetic screening with the zebrafish. In fact, a large collection of mutations that affect nephron formation and function were generated through phenotype-based forward screens. Complementary reverse genetic approaches have also been insightful, with methods spanning the use of antisense morpholino oligonucleotides to genome editing approaches such as the CRISPR/Cas9 system, to selectively knock down or knock out genes of interest to see if they produce kidney phenotypes. Acute kidney injury (AKI) has also been easily modeled in the zebrafish by injecting nephrotoxins, directly inducing damage through surgical intervention, or by generating transgenic lines that express compounds in a tissue-specific manner that when exposed to certain drugs promote an apoptotic response within cells. In this chapter, we provide an overview of these various approaches as well as discuss many of the contributions that have been achieved through the use of zebrafish to model kidney disease.
动物模型一直是推进生物医学研究的宝贵手段，因为它们为疾病病理学的细胞和分子研究提供了实验途径。斑马鱼（Danio rerio）是哺乳动物模型的一个很好的替代选择，可用于应用通常在无脊椎动物中使用的强大遗传实验方法来回答有关脊椎动物发育和疾病的问题。就肾脏而言，斑马鱼已证明自己是一个适用且通用的实验系统，主要是由于其前肾的简单性，前肾包含两个肾单位，这些肾单位在结构和生理方面与哺乳动物的肾单位具有保守性。通过对斑马鱼进行基因筛选，许多以前与肾脏疾病无关的基因现在已与肾脏疾病联系起来。事实上，通过基于表型的正向筛选产生了大量影响肾单位形成和功能的突变。互补的反向遗传方法也很有见地，其方法涵盖从使用反义吗啉代寡核苷酸到基因组编辑方法，如CRISPR/Cas9系统，以选择性地敲低或敲除感兴趣的基因，看看它们是否产生肾脏表型。通过注射肾毒素、通过手术干预直接诱导损伤或通过生成以组织特异性方式表达化合物的转基因品系（当暴露于某些药物时会促进细胞内的凋亡反应），也很容易在斑马鱼中模拟急性肾损伤（AKI）。在本章中，我们概述了这些各种方法，并讨论了通过使用斑马鱼来模拟肾脏疾病所取得的许多成果。
