PUBLICATION

Evaluation of qPCR reference genes in GH-overexpressing transgenic zebrafish (Danio rerio)

Authors
Rassier, G.T., Silveira, T.L.R., Remião, M.H., Daneluz, L.O., Martins, A.W.S., Dellagostin, E.N., Ortiz, H.G., Domingues, W.B., Komninou, E.R., Kütter, M.T., Marins, L.F.F., Campos, V.F.
ID
ZDB-PUB-200731-15
Date
2020
Source
Scientific Reports   10: 12692 (Journal)
Registered Authors
Marins, Luis Fernando
Keywords
none
MeSH Terms
  • Liver/chemistry
  • Brain Chemistry
  • Intestines/chemistry
  • Animals
  • Muscle, Skeletal/chemistry
  • Zebrafish Proteins/genetics*
  • Real-Time Polymerase Chain Reaction/standards*
  • Real-Time Polymerase Chain Reaction/veterinary
  • Zebrafish/genetics*
  • Growth Hormone/genetics*
  • Animals, Genetically Modified
  • Reference Standards
  • Software
(all 13)
PubMed
32728128 Full text @ Sci. Rep.
Abstract
Reference genes (RGs) must have a stable expression in tissues in all experimental conditions to normalize real-time quantitative reverse transcription PCR (qRT-PCR) data. F0104 is a highly studied lineage of zebrafish developed to overexpress the growth hormone (GH). It is assumed that the transgenic process may influence the expression levels of commonly used RGs. The objective of the present study was to make a comprehensive analysis of stability of canditade RGs actb1, actb2, b2m, eif2s2, eef1a1, gapdh, rplp2, rpl7, rpl13α, tuba1, and rps18, in gh-transgenic and non-transgenic zebrafish. Liver, brain, intestine and muscle samples from both groups had qRT-PCR results analyzed by dCt, geNorm, NormFinder, BestKeeper, and RefFinder softwares. Consensus analyses among software concluded that rpl13α, rpl7, and eef1a1 are the most stable genes for zebrafish, considering the studied groups and tissues. Gapdh, rps18, and tuba1 suffered variations in stability among different tissues of both groups, and so, they were listed as the genes with lowest stability. Results from an average pairwise variations test indicated that the use of two RGs would generate reliable results for gene expression analysis in the studied tissues. We conclude that genes that are commonly used in mammals for qRT-PCR assays have low stability in both non-transgenic and gh-transgenic zebrafish reinforcing the importance of using species-specific RGs.
Genes / Markers
Figures
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Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
ufbF0104aTgTransgenic Insertion
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    Human Disease / Model
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    Sequence Targeting Reagents
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    Fish
    No data available
    Antibodies
    No data available
    Orthology
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    Engineered Foreign Genes
    No data available
    Mapping
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