Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.

The analysis of kidney malformation caused by environmental influences during nephrogenesis or by hereditary nephropathies requires animal models allowing the in vivo observation of developmental processes. The zebrafish has emerged as a useful model system for the analysis of vertebrate organ devel...

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Autores principales:	Jens H Westhoff, Stefan Giselbrecht, Miriam Schmidts, Sebastian Schindler, Philip L Beales, Burkhard Tönshoff, Urban Liebel, Jochen Gehrig
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/196d3fdff73e4263a2af9ab55b68685c
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spelling	oai:doaj.org-article:196d3fdff73e4263a2af9ab55b68685c2021-11-18T08:43:28ZDevelopment of an automated imaging pipeline for the analysis of the zebrafish larval kidney.1932-620310.1371/journal.pone.0082137https://doaj.org/article/196d3fdff73e4263a2af9ab55b68685c2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24324758/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The analysis of kidney malformation caused by environmental influences during nephrogenesis or by hereditary nephropathies requires animal models allowing the in vivo observation of developmental processes. The zebrafish has emerged as a useful model system for the analysis of vertebrate organ development and function, and it is suitable for the identification of organotoxic or disease-modulating compounds on a larger scale. However, to fully exploit its potential in high content screening applications, dedicated protocols are required allowing the consistent visualization of inner organs such as the embryonic kidney. To this end, we developed a high content screening compatible pipeline for the automated imaging of standardized views of the developing pronephros in zebrafish larvae. Using a custom designed tool, cavities were generated in agarose coated microtiter plates allowing for accurate positioning and orientation of zebrafish larvae. This enabled the subsequent automated acquisition of stable and consistent dorsal views of pronephric kidneys. The established pipeline was applied in a pilot screen for the analysis of the impact of potentially nephrotoxic drugs on zebrafish pronephros development in the Tg(wt1b:EGFP) transgenic line in which the developing pronephros is highlighted by GFP expression. The consistent image data that was acquired allowed for quantification of gross morphological pronephric phenotypes, revealing concentration dependent effects of several compounds on nephrogenesis. In addition, applicability of the imaging pipeline was further confirmed in a morpholino based model for cilia-associated human genetic disorders associated with different intraflagellar transport genes. The developed tools and pipeline can be used to study various aspects in zebrafish kidney research, and can be readily adapted for the analysis of other organ systems.Jens H WesthoffStefan GiselbrechtMiriam SchmidtsSebastian SchindlerPhilip L BealesBurkhard TönshoffUrban LiebelJochen GehrigPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 12, p e82137 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jens H Westhoff Stefan Giselbrecht Miriam Schmidts Sebastian Schindler Philip L Beales Burkhard Tönshoff Urban Liebel Jochen Gehrig Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.
description	The analysis of kidney malformation caused by environmental influences during nephrogenesis or by hereditary nephropathies requires animal models allowing the in vivo observation of developmental processes. The zebrafish has emerged as a useful model system for the analysis of vertebrate organ development and function, and it is suitable for the identification of organotoxic or disease-modulating compounds on a larger scale. However, to fully exploit its potential in high content screening applications, dedicated protocols are required allowing the consistent visualization of inner organs such as the embryonic kidney. To this end, we developed a high content screening compatible pipeline for the automated imaging of standardized views of the developing pronephros in zebrafish larvae. Using a custom designed tool, cavities were generated in agarose coated microtiter plates allowing for accurate positioning and orientation of zebrafish larvae. This enabled the subsequent automated acquisition of stable and consistent dorsal views of pronephric kidneys. The established pipeline was applied in a pilot screen for the analysis of the impact of potentially nephrotoxic drugs on zebrafish pronephros development in the Tg(wt1b:EGFP) transgenic line in which the developing pronephros is highlighted by GFP expression. The consistent image data that was acquired allowed for quantification of gross morphological pronephric phenotypes, revealing concentration dependent effects of several compounds on nephrogenesis. In addition, applicability of the imaging pipeline was further confirmed in a morpholino based model for cilia-associated human genetic disorders associated with different intraflagellar transport genes. The developed tools and pipeline can be used to study various aspects in zebrafish kidney research, and can be readily adapted for the analysis of other organ systems.
format	article
author	Jens H Westhoff Stefan Giselbrecht Miriam Schmidts Sebastian Schindler Philip L Beales Burkhard Tönshoff Urban Liebel Jochen Gehrig
author_facet	Jens H Westhoff Stefan Giselbrecht Miriam Schmidts Sebastian Schindler Philip L Beales Burkhard Tönshoff Urban Liebel Jochen Gehrig
author_sort	Jens H Westhoff
title	Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.
title_short	Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.
title_full	Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.
title_fullStr	Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.
title_full_unstemmed	Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.
title_sort	development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/196d3fdff73e4263a2af9ab55b68685c
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Development of an automated imaging pipeline for the analysis of the zebrafish larval kidney.

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