PUBLICATION
Evaluation of reference genes for reverse transcription-quantitative PCR assays in organs of zebrafish exposed to glyphosate-based herbicide, Roundup
- Authors
- Jaramillo, M.L., Pereira, A.G., Davico, C.E., Nezzi, L., Ammar, D., Müller, Y.M.R., Nazari, E.M.
- ID
- ZDB-PUB-171128-6
- Date
- 2017
- Source
- Animal : an international journal of animal bioscience 12(7): 1424-1434 (Journal)
- Registered Authors
- Keywords
- Danio rerio, Roundup WG, expression stability, normalization, reverse transcription-quantitative PCR
- MeSH Terms
-
- Animals
- Female
- Gene Expression Profiling*
- Glycine/analogs & derivatives*
- Glycine/toxicity
- Herbicides
- Male
- Real-Time Polymerase Chain Reaction*
- Reference Standards
- Reverse Transcription
- Zebrafish*/genetics
- PubMed
- 29173213 Full text @ Animal
Citation
Jaramillo, M.L., Pereira, A.G., Davico, C.E., Nezzi, L., Ammar, D., Müller, Y.M.R., Nazari, E.M. (2017) Evaluation of reference genes for reverse transcription-quantitative PCR assays in organs of zebrafish exposed to glyphosate-based herbicide, Roundup. Animal : an international journal of animal bioscience. 12(7):1424-1434.
Abstract
Roundup is a glyphosate-based herbicide (GBH) widely used in agriculture and may cause toxic effects in non-target organisms. Model organisms, as zebrafish, and analysis of gene expression by reverse transcription-quantitative PCR (RT-qPCR) could be used to better understand the Roundup toxicity. A prerequisite for RT-qPCR is the availability of appropriate reference genes; however, they have not been described for Roundup-exposed fish. The aim of this study was to evaluate the expression stability of six reference genes (rpl8, β-act, gapdh, b2m, ef1α, hprt1) and one expressed repetitive element (hatn10) in organs of males (brain, gill, testis) and females (ovary) of zebrafish exposed to Roundup WG at three concentrations (0.065, 0.65 and 6.5 mg N-(phosphonomethyl) glycine/l) for 7 days. Genes were ranked by geNorm, NormFinder, BestKeeper, Delta C t and RefFinder, and their best combinations were determined by geNorm and NormFinder programs. The two most stable ranked genes were specific to each organ: gill (β-act; rpl8); brain (rpl8; β-act); testis (ef1α; gapdh); and ovary (rpl8; hprt1). The cat transcript level was used to evaluate the effect of normalization with these reference genes. These are the first suitable reference genes described for the analysis of gene expression in organs of Roundup-exposed zebrafish, and will allow investigations of the molecular mechanisms of Roundup toxicity.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping