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Open AccessArticle

A Multiparametric Assay Platform for Simultaneous In Vivo Assessment of Pronephric Morphology, Renal Function and Heart Rate in Larval Zebrafish

1
Department of Pediatrics I, University Children’s Hospital Heidelberg, 69120 Heidelberg, Germany
2
DITABIS, Digital Biomedical Imaging Systems AG, 75179 Pforzheim, Germany
3
ACQUIFER Imaging GmbH, 69123 Heidelberg, Germany
*
Authors to whom correspondence should be addressed.
The authors contributed equally.
Present address: Developmental Biology Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
Cells 2020, 9(5), 1269; https://doi.org/10.3390/cells9051269
Received: 1 April 2020 / Revised: 13 May 2020 / Accepted: 18 May 2020 / Published: 20 May 2020
(This article belongs to the Special Issue Signaling Pathway Analysis and Disease Modeling in Zebrafish)
Automated high-throughput workflows allow for chemical toxicity testing and drug discovery in zebrafish disease models. Due to its conserved structural and functional properties, the zebrafish pronephros offers a unique model to study renal development and disease at larger scale. Ideally, scoring of pronephric phenotypes includes morphological and functional assessments within the same larva. However, to efficiently upscale such assays, refinement of existing methods is required. Here, we describe the development of a multiparametric in vivo screening pipeline for parallel assessment of pronephric morphology, kidney function and heart rate within the same larva on a single imaging platform. To this end, we developed a novel 3D-printed orientation tool enabling multiple consistent orientations of larvae in agarose-filled microplates. Dorsal pronephros imaging was followed by assessing renal clearance and heart rates upon fluorescein isothiocyanate (FITC)-inulin microinjection using automated time-lapse imaging of laterally positioned larvae. The pipeline was benchmarked using a set of drugs known to induce developmental nephrotoxicity in humans and zebrafish. Drug-induced reductions in renal clearance and heart rate alterations were detected even in larvae exhibiting minor pronephric phenotypes. In conclusion, the developed workflow enables rapid and semi-automated in vivo assessment of multiple morphological and functional parameters. View Full-Text
Keywords: zebrafish; automated imaging; screening; developmental toxicity; pronephros; kidney function; kidney morphology; heart rate zebrafish; automated imaging; screening; developmental toxicity; pronephros; kidney function; kidney morphology; heart rate
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MDPI and ACS Style

Steenbergen, P.J.; Heigwer, J.; Pandey, G.; Tönshoff, B.; Gehrig, J.; Westhoff, J.H. A Multiparametric Assay Platform for Simultaneous In Vivo Assessment of Pronephric Morphology, Renal Function and Heart Rate in Larval Zebrafish. Cells 2020, 9, 1269. https://doi.org/10.3390/cells9051269

AMA Style

Steenbergen PJ, Heigwer J, Pandey G, Tönshoff B, Gehrig J, Westhoff JH. A Multiparametric Assay Platform for Simultaneous In Vivo Assessment of Pronephric Morphology, Renal Function and Heart Rate in Larval Zebrafish. Cells. 2020; 9(5):1269. https://doi.org/10.3390/cells9051269

Chicago/Turabian Style

Steenbergen, Petrus J.; Heigwer, Jana; Pandey, Gunjan; Tönshoff, Burkhard; Gehrig, Jochen; Westhoff, Jens H. 2020. "A Multiparametric Assay Platform for Simultaneous In Vivo Assessment of Pronephric Morphology, Renal Function and Heart Rate in Larval Zebrafish" Cells 9, no. 5: 1269. https://doi.org/10.3390/cells9051269

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