Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models

Abstract The majority of kidney diseases arise from the loss of podocytes and from morphological changes of their highly complex foot process architecture, which inevitably leads to a reduced kidney filtration and total loss of kidney function. It could have been shown that microRNAs (miRs) play a p...

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Autores principales:	Florian Siegerist, Tim Lange, Anna Iervolino, Thor Magnus Koppe, Weibin Zhou, Giovambattista Capasso, Karlhans Endlich, Nicole Endlich
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/540c83e1c7534d7dbdac41cae1c862dd
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spelling	oai:doaj.org-article:540c83e1c7534d7dbdac41cae1c862dd2021-11-28T12:21:52ZEvaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models10.1038/s41598-021-00075-22045-2322https://doaj.org/article/540c83e1c7534d7dbdac41cae1c862dd2021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-00075-2https://doaj.org/toc/2045-2322Abstract The majority of kidney diseases arise from the loss of podocytes and from morphological changes of their highly complex foot process architecture, which inevitably leads to a reduced kidney filtration and total loss of kidney function. It could have been shown that microRNAs (miRs) play a pivotal role in the pathogenesis of podocyte-associated kidney diseases. Due to their fully functioning pronephric kidney, larval zebrafish have become a popular vertebrate model, to study kidney diseases in vivo. Unfortunately, there is no consensus about a proper normalization strategy of RT-qPCR-based miRNA expression data in zebrafish. In this study we analyzed 9 preselected candidates dre-miR-92a-3p, dre-miR-206-3p, dre-miR-99-1, dre-miR-92b-3p, dre-miR-363-3p, dre-let-7e, dre-miR-454a, dre-miR-30c-5p, dre-miR-126a-5p for their capability as endogenous reference genes in zebrafish experiments. Expression levels of potential candidates were measured in 3 different zebrafish strains, different developmental stages, and in different kidney disease models by RT-qPCR. Expression values were analyzed with NormFinder, BestKeeper, GeNorm, and DeltaCt and were tested for inter-group differences. All candidates show an abundant expression throughout all samples and relatively high stability. The most stable candidate without significant inter-group differences was dre-miR-92b-3p making it a suitable endogenous reference gene for RT-qPCR-based miR expression zebrafish studies.Florian SiegeristTim LangeAnna IervolinoThor Magnus KoppeWeibin ZhouGiovambattista CapassoKarlhans EndlichNicole EndlichNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Florian Siegerist Tim Lange Anna Iervolino Thor Magnus Koppe Weibin Zhou Giovambattista Capasso Karlhans Endlich Nicole Endlich Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models
description	Abstract The majority of kidney diseases arise from the loss of podocytes and from morphological changes of their highly complex foot process architecture, which inevitably leads to a reduced kidney filtration and total loss of kidney function. It could have been shown that microRNAs (miRs) play a pivotal role in the pathogenesis of podocyte-associated kidney diseases. Due to their fully functioning pronephric kidney, larval zebrafish have become a popular vertebrate model, to study kidney diseases in vivo. Unfortunately, there is no consensus about a proper normalization strategy of RT-qPCR-based miRNA expression data in zebrafish. In this study we analyzed 9 preselected candidates dre-miR-92a-3p, dre-miR-206-3p, dre-miR-99-1, dre-miR-92b-3p, dre-miR-363-3p, dre-let-7e, dre-miR-454a, dre-miR-30c-5p, dre-miR-126a-5p for their capability as endogenous reference genes in zebrafish experiments. Expression levels of potential candidates were measured in 3 different zebrafish strains, different developmental stages, and in different kidney disease models by RT-qPCR. Expression values were analyzed with NormFinder, BestKeeper, GeNorm, and DeltaCt and were tested for inter-group differences. All candidates show an abundant expression throughout all samples and relatively high stability. The most stable candidate without significant inter-group differences was dre-miR-92b-3p making it a suitable endogenous reference gene for RT-qPCR-based miR expression zebrafish studies.
format	article
author	Florian Siegerist Tim Lange Anna Iervolino Thor Magnus Koppe Weibin Zhou Giovambattista Capasso Karlhans Endlich Nicole Endlich
author_facet	Florian Siegerist Tim Lange Anna Iervolino Thor Magnus Koppe Weibin Zhou Giovambattista Capasso Karlhans Endlich Nicole Endlich
author_sort	Florian Siegerist
title	Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models
title_short	Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models
title_full	Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models
title_fullStr	Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models
title_full_unstemmed	Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models
title_sort	evaluation of endogenous mirna reference genes across different zebrafish strains, developmental stages and kidney disease models
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/540c83e1c7534d7dbdac41cae1c862dd
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Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models

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