title: A Fluorescence-Based Assay for Proteinuria Screening in Larval Zebrafish (Danio rerio).
abstract: Analysis of genes compromising the glomerular filtration barrier in rodent models using transgenic or knockdown approaches is time- and resource-consuming and often leads to unsatisfactory results. Therefore, it would be beneficial to have a selection tool indicating that your gene of interest is in fact associated with proteinuria. Zebrafish (Danio rerio) is a rapid screening tool to study effects in glomerular filtration barrier integrity after genetic manipulation. We use either injection of high-molecular-weight dextrans or a transgenic fluorescent fish line [Tg(l-fabp:DBP:EGFP)] expressing a vitamin D-binding protein fused with eGFP for indirect detection of proteinuria. A loss of high-molecular-weight proteins from the circulation of the fish into the urine can be identified by monitoring fluorescence intensity in the zebrafish eye. Paired with an optimized analysis method, this assay provides an effective screening solution to detect filtration barrier damage with proteinuria before moving to a mammalian system.
authors.affiliation: 1 Division of Nephrology, Hannover Medical School , Hannover, Germany .
authors.forename: Nils
authors.initials: N
authors.lastname: Hanke
authors.affiliation: 2 Mount Desert Island Biological Laboratory , Bar Harbor, Maine.
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: King
authors.affiliation: 3 Division of Biometry, Hannover Medical School , Hannover, Germany .
authors.forename: Bernhard
authors.initials: B
authors.lastname: Vaske
authors.affiliation: 1 Division of Nephrology, Hannover Medical School , Hannover, Germany .|2 Mount Desert Island Biological Laboratory , Bar Harbor, Maine.
authors.forename: Hermann
authors.initials: H
authors.lastname: Haller
authors.affiliation: 1 Division of Nephrology, Hannover Medical School , Hannover, Germany .|2 Mount Desert Island Biological Laboratory , Bar Harbor, Maine.
authors.forename: Mario
authors.initials: M
authors.lastname: Schiffer
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers; D003911:Dextrans / Q000378:metabolism*; D005123:Eye / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D023421:Models, Animal; D011507:Proteinuria / Q000235:genetics*; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D003911:Dextrans; D049452:Green Fluorescent Proteins