Automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions

Abstract Accurate quantification of heartbeats in fish models is an important readout to study cardiovascular biology, disease states and pharmacology. However, dependence on anaesthesia, laborious sample orientation or requirement for fluorescent reporters have hampered the use of high-throughput h...

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Autores principales:	Jakob Gierten, Christian Pylatiuk, Omar T. Hammouda, Christian Schock, Johannes Stegmaier, Joachim Wittbrodt, Jochen Gehrig, Felix Loosli
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e7599b73afdf4766a3526d1359de26d9
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spelling	oai:doaj.org-article:e7599b73afdf4766a3526d1359de26d92021-12-02T14:06:58ZAutomated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions10.1038/s41598-020-58563-w2045-2322https://doaj.org/article/e7599b73afdf4766a3526d1359de26d92020-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-58563-whttps://doaj.org/toc/2045-2322Abstract Accurate quantification of heartbeats in fish models is an important readout to study cardiovascular biology, disease states and pharmacology. However, dependence on anaesthesia, laborious sample orientation or requirement for fluorescent reporters have hampered the use of high-throughput heartbeat analysis. To overcome these limitations, we established an efficient screening assay employing automated label-free heart rate determination of randomly oriented, non-anesthetized medaka (Oryzias latipes) and zebrafish (Danio rerio) embryos in microtiter plates. Automatically acquired bright-field data feeds into an easy-to-use HeartBeat software with graphical user interface for automated quantification of heart rate and rhythm. Sensitivity of the assay was demonstrated by profiling heart rates during entire embryonic development. Our analysis revealed rapid adaption of heart rates to temperature changes, which has implications for standardization of experimental layout. The assay allows scoring of multiple embryos per well enabling a throughput of >500 embryos per 96-well plate. In a proof of principle screen for compound testing, we captured concentration-dependent effects of nifedipine and terfenadine over time. Our novel assay permits large-scale applications ranging from phenotypic screening, interrogation of gene functions to cardiovascular drug development.Jakob GiertenChristian PylatiukOmar T. HammoudaChristian SchockJohannes StegmaierJoachim WittbrodtJochen GehrigFelix LoosliNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-12 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jakob Gierten Christian Pylatiuk Omar T. Hammouda Christian Schock Johannes Stegmaier Joachim Wittbrodt Jochen Gehrig Felix Loosli Automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions
description	Abstract Accurate quantification of heartbeats in fish models is an important readout to study cardiovascular biology, disease states and pharmacology. However, dependence on anaesthesia, laborious sample orientation or requirement for fluorescent reporters have hampered the use of high-throughput heartbeat analysis. To overcome these limitations, we established an efficient screening assay employing automated label-free heart rate determination of randomly oriented, non-anesthetized medaka (Oryzias latipes) and zebrafish (Danio rerio) embryos in microtiter plates. Automatically acquired bright-field data feeds into an easy-to-use HeartBeat software with graphical user interface for automated quantification of heart rate and rhythm. Sensitivity of the assay was demonstrated by profiling heart rates during entire embryonic development. Our analysis revealed rapid adaption of heart rates to temperature changes, which has implications for standardization of experimental layout. The assay allows scoring of multiple embryos per well enabling a throughput of >500 embryos per 96-well plate. In a proof of principle screen for compound testing, we captured concentration-dependent effects of nifedipine and terfenadine over time. Our novel assay permits large-scale applications ranging from phenotypic screening, interrogation of gene functions to cardiovascular drug development.
format	article
author	Jakob Gierten Christian Pylatiuk Omar T. Hammouda Christian Schock Johannes Stegmaier Joachim Wittbrodt Jochen Gehrig Felix Loosli
author_facet	Jakob Gierten Christian Pylatiuk Omar T. Hammouda Christian Schock Johannes Stegmaier Joachim Wittbrodt Jochen Gehrig Felix Loosli
author_sort	Jakob Gierten
title	Automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions
title_short	Automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions
title_full	Automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions
title_fullStr	Automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions
title_full_unstemmed	Automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions
title_sort	automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/e7599b73afdf4766a3526d1359de26d9
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Automated high-throughput heartbeat quantification in medaka and zebrafish embryos under physiological conditions

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