RT Journal Article
SR Electronic
T1 Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.04.09.439067
DO 10.1101/2021.04.09.439067
A1 Florian Siegerist
A1 Tim Lange
A1 Anna Iervolino
A1 Thor Magnus Koppe
A1 Weibin Zhou
A1 Giovambattista Capasso
A1 Karlhans Endlich
A1 Nicole Endlich
YR 2021
UL http://biorxiv.org/content/early/2021/04/10/2021.04.09.439067.abstract
AB 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.Competing Interest StatementThe authors have declared no competing interest.