Department of Medicine 4 - Nephrology and Hypertension, Universitätsklinikum Erlangen, Erlangen, Germany.
Friedrich-Alexander Universiät Erlangen-Nuremberg, Erlangen, Germany.
Methods Mol Biol. 2023;2664:145-157. doi: 10.1007/978-1-0716-3179-9_11.
The zebrafish model has been used in many different fields of research because of its high homology to the human genome, its easy genetic manipulation, its high fecundity, and its rapid development. For glomerular diseases, zebrafish larvae have proven to be a versatile tool to study the contribution of different genes, because the zebrafish pronephros is very comparable to the human kidney in function and ultrastructure. Here we describe the principle and use of a simple screening assay based on the measurement of the fluorescence in the retinal vessel plexus of the Tg(l-fabp:DBP:eGFP) zebrafish line ("eye assay") to indirectly determine proteinuria as a hallmark of podocyte dysfunction. Furthermore, we illustrate how to analyze the obtained data and outline methods to attribute the findings to podocyte impairment.
斑马鱼模型由于与人类基因组具有高度同源性、易于基因操作、繁殖能力强、发育迅速，已被广泛应用于多个研究领域。对于肾小球疾病，斑马鱼幼虫已被证明是研究不同基因贡献的多功能工具，因为斑马鱼前肾在功能和超微结构上与人类肾脏非常相似。在这里，我们描述了一种基于 Tg(l-fabp:DBP:eGFP) 斑马鱼系视网膜血管丛荧光测量的简单筛选测定法的原理和用途（“眼测定法”），以间接确定蛋白尿作为足细胞功能障碍的标志。此外，我们还说明了如何分析获得的数据，并概述了将这些发现归因于足细胞损伤的方法。
