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.

journal_name

Zebrafish

journal_title

Zebrafish

authors

Hanke N,King BL,Vaske B,Haller H,Schiffer M

doi

10.1089/zeb.2015.1093

subject

Has Abstract

pub_date

2015-10-01 00:00:00

pages

372-6

issue

5

eissn

1545-8547

issn

1557-8542

journal_volume

12

pub_type

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