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
  • Gene Expression Regulation, Developmental
  • Genotype
  • Disease Models, Animal
  • Podocytes/metabolism
  • Podocytes/pathology
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism
  • MicroRNAs/genetics*
  • MicroRNAs/metabolism
  • Kidney Diseases/genetics*
  • Kidney Diseases/metabolism
  • Kidney Diseases/pathology
  • Real-Time Polymerase Chain Reaction
  • Animals, Genetically Modified
  • Animals
  • Larva/genetics
  • Larva/metabolism
  • Phenotype
  • Reverse Transcriptase Polymerase Chain Reaction
  • Gene Knockdown Techniques
(all 21)
PubMed
34819534 Full text @ Sci. Rep.
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
Figures
Figure Gallery (7 images)
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Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
a9
    		Complex
    mi1000TgTransgenic Insertion
      mi1004TgTransgenic Insertion
        rw0144TgTransgenic Insertion
          w2
            		Point Mutation
            1 - 5 of 5
            Show
            Human Disease / Model
            No data available
            Sequence Targeting Reagents
            Target Reagent Reagent Type
            nphs1MO1-nphs1MRPHLNO
            wt1aMO3-wt1aMRPHLNO
            1 - 2 of 2
            Show
            Fish
            No data available
            Antibodies
            No data available
            Orthology
            No data available
            Engineered Foreign Genes
            Marker Marker Type Name
            EGFPEFGEGFP
            GAL4EFGGAL4
            mCherryEFGmCherry
            1 - 3 of 3
            Show
            Mapping
            No data available
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            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.