Abstract
With the advances in next-generation sequencing and rapid filtering of candidate variants in diseased patients, it has been increasingly important to develop translatable in vivo models to study genetic changes. This allows for functional validation of pathogenic mutations and establishes a system to understand the etiology of disease. Due to the ease of genetic manipulation and rapid ex utero development, the zebrafish has become a valuable resource to study important biological processes, including nephrogenesis. The development and function of the zebrafish pronephros are akin to that of mammals. As such, they offer a tractable model to study kidney disease, especially diabetic nephropathy. However, in order to study kidney dysfunction in zebrafish it is imperative that an appropriate readout is available. The appearance of macro-proteins in patient’s urine is indicative of defective kidney function. In this technical chapter, we describe the in vivo use of fluorescently tagged dextrans of different molecular weights to reveal the integrity of the zebrafish glomerular filtration barrier.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Outtandy P, Russell C, Kleta R, Bockenhauer D (2019) Zebrafish as a model for kidney function and disease. Pediatr Nephrol 34:751–762
Drummond I (2003) Making a zebrafish kidney: a tale of two tubes. Trends Cell Biol 13:357–365
Drummond IA, Majumdar A, Hentschel H, Elger M, Solnica-Krezel L, Schier AF, Neuhauss SC, Stemple DL, Zwartkruis F, Rangini Z, Driever W, Fishman MC (1998) Early development of the zebrafish pronephros and analysis of mutations affecting pronephric function. Development 125:4655–4667
Gerlach GF, Wingert RA (2013) Kidney organogenesis in the zebrafish: insights into vertebrate nephrogenesis and regeneration. Wiley Interdiscip Rev Dev Biol 2:559–585
Wingert RA, Davidson AJ (2008) The zebrafish pronephros: a model to study nephron segmentation. Kidney Int 73:1120–1127
Ichimura K, Bubenshchikova E, Powell R, Fukuyo Y, Nakamura T, Tran U, Oda S, Tanaka M, Wessely O, Kurihara H, Sakai T, Obara T (2012) A comparative analysis of glomerulus development in the pronephros of medaka and zebrafish. PLoS One 7:e45286
Fukuyo Y, Nakamura T, Bubenshchikova E, Powell R, Tsuji T, Janknecht R, Obara T (2014) Nephrin and Podocin functions are highly conserved between the zebrafish pronephros and mammalian metanephros. Mol Med Rep 9:457–465
Christou-Savina S, Beales PL, Osborn DP (2015) Evaluation of zebrafish kidney function using a fluorescent clearance assay. J Vis Exp:e52540
Reidy K, Kang HM, Hostetter T, Susztak K (2014) Molecular mechanisms of diabetic kidney disease. J Clin Invest 124:2333–2340
Dorsemans AC, Lefebvre d’Hellencourt C, Ait-Arsa I, Jestin E, Meilhac O, Diotel N (2017) Acute and chronic models of hyperglycemia in zebrafish: a method to assess the impact of hyperglycemia on neurogenesis and the biodistribution of radiolabeled molecules. J Vis Exp:e55203
Powers JW, Mazilu JK, Lin S, McCabe ER (2010) The effects of hyperglycemia on adrenal cortex function and steroidogenesis in the zebrafish. Mol Genet Metab 101:421–422
Elo B, Villano CM, Govorko D, White LA (2007) Larval zebrafish as a model for glucose metabolism: expression of phosphoenolpyruvate carboxykinase as a marker for exposure to anti-diabetic compounds. J Mol Endocrinol 38:433–440
Jörgens K, Hillebrands JL, Hammes HP, Kroll J (2012) Zebrafish: a model for understanding diabetic complications. Exp Clin Endocrinol Diabetes 120:186–187
Gleeson M, Connaughton V, Arneson LS (2007) Induction of hyperglycaemia in zebrafish (Danio rerio) leads to morphological changes in the retina. Acta Diabetol 44:157–163
Alvarez Y, Chen K, Reynolds AL, Waghorne N, O’Connor JJ, Kennedy BN (2010) Predominant cone photoreceptor dysfunction in a hyperglycaemic model of non-proliferative diabetic retinopathy. Dis Model Mech 3:236–245
Carnovali M, Luzi L, Banfi G, Mariotti M (2016) Chronic hyperglycemia affects bone metabolism in adult zebrafish scale model. Endocrine 54:808–817
Zang L, Maddison LA, Chen W (2018) Zebrafish as a model for obesity and diabetes. Front Cell Dev Biol 6:91
Olsen AS, Sarras MP, Jr RVI (2010) Limb regeneration is impaired in an adult zebrafish model of diabetes mellitus. Wound Repair Regen 18:532–542
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kolatsi-Joannou, M., Osborn, D. (2020). A Technique for Studying Glomerular Filtration Integrity in the Zebrafish Pronephros. In: Gnudi, L., Long, D. (eds) Diabetic Nephropathy. Methods in Molecular Biology, vol 2067. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9841-8_3
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9841-8_3
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9840-1
Online ISBN: 978-1-4939-9841-8
eBook Packages: Springer Protocols