PUBLICATION
Validation of Housekeeping Genes as Internal Controls for the Study of the Effects of Microcystin-LR in Zebrafish by Real-Time PCR
- Authors
- Sopezki, M.D.S., Monserrat, J.M., Yunes, J.S., Zanette, J.
- ID
- ZDB-PUB-180726-5
- Date
- 2018
- Source
- Zebrafish 15(5): 454-459 (Journal)
- Registered Authors
- Keywords
- Danio rerio, MC-LR, RT-qPCR, reference gene
- MeSH Terms
-
- Animals
- Enzyme Inhibitors/pharmacology*
- Gene Expression Regulation/drug effects*
- Genes, Essential*
- Liver/drug effects
- Liver/metabolism*
- Microcystins/pharmacology*
- Real-Time Polymerase Chain Reaction/methods*
- Real-Time Polymerase Chain Reaction/standards
- Reference Standards
- Zebrafish
- PubMed
- 30044200 Full text @ Zebrafish
Citation
Sopezki, M.D.S., Monserrat, J.M., Yunes, J.S., Zanette, J. (2018) Validation of Housekeeping Genes as Internal Controls for the Study of the Effects of Microcystin-LR in Zebrafish by Real-Time PCR. Zebrafish. 15(5):454-459.
Abstract
Microcystin-leucine arginine (MC-LR) is a natural toxin produced by cyanobacterial blooms that causes severe liver damage in fish. It is crucial to investigate if housekeeping genes are affected by MC-LR in zebrafish, to permit the adequate evaluation of gene expression by RT-qPCR. We evaluated the gene expression stability (M value) and regulation by chemical treatment (using E-Ct and E-ΔCt) to validate the use of eight housekeeping genes in fish exposed to 0, 0.31, and 6.10 μg L-1 MC-LR for 24 h. We suggest the use of the combination of β-actin1, b2m, and arnt2 in the liver and β-actin1, 18S rRNA, and arnt2 in gills as housekeeping genes. The evaluation of gene regulation following MC-LR exposure denoted a strong repression of 18S rRNA (17- and 10-fold decrease) and tbp (10- and 2-fold decrease) and induction of ef-1α (8- and 14-fold increase) in the liver of zebrafish exposed to 0.31 and 6.10 μg L-1 MC-LR, respectively. This is the first study showing that housekeeping genes commonly used in gene expression could be affected in the liver by environmentally relevant concentrations of MC-LR. The study validates adequate housekeeping genes that could be used in toxicological studies with MC-LR in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping