Transgenic zebrafish modeling low-molecular-weight proteinuria and lysosomal storage diseases
Chen, Zhiyong; Luciani, Alessandro; Mateos, José María; Barmettler, Gery; Giles, Rachel H.; Neuhauss, Stephan C.F.; Devuyst, Olivier
(2020) Kidney International, volume 97, issue 6, pp. 1150 - 1163
(Article)
Abstract
Epithelial cells lining the proximal tubule of the kidney reabsorb and metabolize most of the filtered low-molecular-weight proteins through receptor-mediated endocytosis and lysosomal processing. Congenital and acquired dysfunctions of the proximal tubule are consistently reflected by the inappropriate loss of solutes including low-molecular-weight proteins in the urine. The zebrafish pronephros
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shares individual functional segments with the human nephron, including lrp2a/megalin-dependent endocytic transport processes of the proximal tubule. Although the zebrafish has been used as a model organism for toxicological studies and drug discovery, there is no available assay that allows large-scale assessment of proximal tubule function in larval or adult stages. Here we establish a transgenic Tg(lfabp::½vdbp-mCherry) zebrafish line expressing in the liver the N-terminal region of vitamin D-binding protein coupled to the acid-insensitive, red monomeric fluorescent protein mCherry (½vdbp-mCherry). This low-molecular-weight protein construct is secreted into the bloodstream, filtered through the glomerulus, reabsorbed by receptor-mediated endocytosis and processed in the lysosomes of proximal tubule cells of the fish. Thus, our proof-of-concept studies using zebrafish larvae knockout for lrp2a and clcn7 or exposed to known nephrotoxins (gentamicin and cisplatin) demonstrate that this transgenic line is useful to monitor low-molecular-weight proteinuria and lysosomal processing. This represents a powerful new model organism for drug screening and studies of nephrotoxicity.
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Keywords: endocytosis, lysosome, model organism, proximal tubule, renal Fanconi syndrome, Nephrology
ISSN: 0085-2538
Publisher: Nature Publishing Group
Note: Funding Information: This project has received funding from the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement No. 608847. We are grateful to ERA-EDTA (research fellowship to ZC), the Swiss National Science Foundation (project grant 31003A-169850), RADIZ (Rare Disease Initiative Zurich) of the UZH, the Cystinosis Research Foundation (Irvine, CA), and the Dutch Kidney Foundation (16OI06). We acknowledge Marine Berquez, Huguette Debaix, Dr. Dominik Hänni, Dr. Christina Pickel, Patrick Spielmann, and Dr. Glenn van de Hoek for providing technical assistance. We thank the zebrafish caretaker team of the Hubrecht Institute and of the zebrafish facility at the University of Zurich. Imaging was performed with equipment maintained by the Center for Microscopy and Image Analysis, University of Zurich. Funding Information: This project has received funding from the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement No. 608847 . We are grateful to ERA-EDTA (research fellowship to ZC), the Swiss National Science Foundation (project grant 31003A-169850 ), RADIZ (Rare Disease Initiative Zurich) of the UZH , the Cystinosis Research Foundation (Irvine, CA), and the Dutch Kidney Foundation ( 16OI06 ). We acknowledge Marine Berquez, Huguette Debaix, Dr. Dominik Hänni, Dr. Christina Pickel, Patrick Spielmann, and Dr. Glenn van de Hoek for providing technical assistance. We thank the zebrafish caretaker team of the Hubrecht Institute and of the zebrafish facility at the University of Zurich. Imaging was performed with equipment maintained by the Center for Microscopy and Image Analysis, University of Zurich. Publisher Copyright: © 2019 International Society of Nephrology
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