A novel nanoluciferase transgenic reporter measures proteinuria in zebrafish.
Alport syndrome
basement membrane
glomerulus
proteinuria
proximal tubule
type IV collagen
zebrafish
Journal
Kidney international
ISSN: 1523-1755
Titre abrégé: Kidney Int
Pays: United States
ID NLM: 0323470
Informations de publication
Date de publication:
10 2022
10 2022
Historique:
received:
20
08
2021
revised:
25
04
2022
accepted:
09
05
2022
pubmed:
19
6
2022
medline:
28
9
2022
entrez:
18
6
2022
Statut:
ppublish
Résumé
The zebrafish is an important animal system for modeling human diseases. This includes kidney dysfunction as the embryonic kidney (pronephros) shares considerable molecular and morphological homology with the human nephron. A key clinical indicator of kidney disease is proteinuria, but a high-throughput readout of proteinuria in the zebrafish is currently lacking. To remedy this, we used the Tol2 transposon system to generate a transgenic zebrafish line that uses the fabp10a liver-specific promoter to over-express a nanoluciferase molecule fused with the D3 domain of Receptor-Associated Protein (a type of molecular chaperone) which we term NL-D3. Using a luminometer, we quantified proteinuria in NL-D3 zebrafish larvae by measuring the intensity of luminescence in the embryo medium. In the healthy state, NL-D3 is not excreted, but when embryos were treated with chemicals that affected either proximal tubular reabsorption (cisplatin, gentamicin) or glomerular filtration (angiotensin II, Hanks Balanced Salt Solution, Bovine Serum Albumin), NL-D3 is detected in fish medium. Similarly, depletion of several gene products associated with kidney disease (nphs1, nphs2, lrp2a, ocrl, col4a3, and col4a4) also induced NL-D3 proteinuria. Treating col4a4 depleted zebrafish larvae (a model of Alport syndrome) with captopril reduced proteinuria in this system. Thus, our findings validate the use of the NL-D3 transgenic zebrafish as a robust and quantifiable proteinuria reporter. Hence, given the feasibility of high-throughput assays in zebrafish, this novel reporter will permit screening for drugs that ameliorate proteinuria, thereby prioritizing candidates for further translational studies.
Identifiants
pubmed: 35716957
pii: S0085-2538(22)00452-5
doi: 10.1016/j.kint.2022.05.019
pmc: PMC7614274
mid: EMS170654
pii:
doi:
Substances chimiques
Gentamicins
0
Angiotensin II
11128-99-7
Serum Albumin, Bovine
27432CM55Q
Captopril
9G64RSX1XD
Cisplatin
Q20Q21Q62J
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
815-827Subventions
Organisme : Wellcome Trust
ID : 202860
Pays : United Kingdom
Informations de copyright
Crown Copyright © 2022. Published by Elsevier Inc. All rights reserved.
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