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
-
- Animals
- Animals, Genetically Modified
- Brain Chemistry
- Growth Hormone/genetics*
- Intestines/chemistry
- Liver/chemistry
- Muscle, Skeletal/chemistry
- Real-Time Polymerase Chain Reaction/standards*
- Real-Time Polymerase Chain Reaction/veterinary
- Reference Standards
- Software
- Zebrafish/genetics*
- Zebrafish Proteins/genetics*
- PubMed
- 32728128 Full text @ Sci. Rep.
Citation
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. (2020) Evaluation of qPCR reference genes in GH-overexpressing transgenic zebrafish (Danio rerio). Scientific Reports. 10:12692.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping