PUBLICATION
Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models
- Authors
- Siegerist, F., Lange, T., Iervolino, A., Koppe, T.M., Zhou, W., Capasso, G., Endlich, K., Endlich, N.
- ID
- ZDB-PUB-211129-22
- Date
- 2021
- Source
- Scientific Reports 11: 22894 (Journal)
- Registered Authors
- Zhou, Weibin
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Disease Models, Animal
- Gene Expression Regulation, Developmental
- Gene Knockdown Techniques
- Genotype
- Kidney Diseases/genetics*
- Kidney Diseases/metabolism
- Kidney Diseases/pathology
- Larva/genetics
- Larva/metabolism
- MicroRNAs/genetics*
- MicroRNAs/metabolism
- Phenotype
- Podocytes/metabolism
- Podocytes/pathology
- Real-Time Polymerase Chain Reaction
- Reverse Transcriptase Polymerase Chain Reaction
- Zebrafish/embryology
- Zebrafish/genetics*
- Zebrafish/metabolism
- PubMed
- 34819534 Full text @ Sci. Rep.
Citation
Siegerist, F., Lange, T., Iervolino, A., Koppe, T.M., Zhou, W., Capasso, G., Endlich, K., Endlich, N. (2021) Evaluation of endogenous miRNA reference genes across different zebrafish strains, developmental stages and kidney disease models. Scientific Reports. 11:22894.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping