Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress
Background One important physiological response to environmental stress in animals is change in gene expression. To obtain reliable data from gene expression studies using RT-qPCR it is important to evaluate a set of possible reference genes as normalizers for expression. The expression of these can...
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ftoslouniv:oai:www.duo.uio.no:10852/46293 2023-05-15T15:27:04+02:00 Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress Aursnes, Ingrid A Rishovd, Anne L Karlsen, Hans E Gjøen, Tor 2011 http://hdl.handle.net/10852/46293 http://urn.nb.no/URN:NBN:no-50541 https://doi.org/10.1186/1756-0500-4-104 eng eng http://urn.nb.no/URN:NBN:no-50541 BMC Research Notes. 2011 Apr 05;4(1):104 http://hdl.handle.net/10852/46293 http://dx.doi.org/10.1186/1756-0500-4-104 URN:NBN:no-50541 Fulltext https://www.duo.uio.no/bitstream/handle/10852/46293/1/13104_2010_Article_856.pdf Gjøen et al; licensee BioMed Central Ltd. Attribution 2.0 Generic http://creativecommons.org/licenses/by/2.0/ CC-BY Journal article Tidsskriftartikkel Peer reviewed PublishedVersion 2011 ftoslouniv https://doi.org/10.1186/1756-0500-4-104 2020-06-21T08:48:49Z Background One important physiological response to environmental stress in animals is change in gene expression. To obtain reliable data from gene expression studies using RT-qPCR it is important to evaluate a set of possible reference genes as normalizers for expression. The expression of these candidate genes should be analyzed in the relevant tissues during normal and stressed situations. To find suitable reference genes it was crucial that the genes were stably expressed also during a situation of physiological stress. For poikilotermic animals like cod, changes in temperature are normal, but if the changes are faster than physiological compensation, the animals respond with typical stress responses. It has previously been shown that Atlantic cod show stress responses when elevation of water temperature is faster than 1 degree/day, for this reason we chose hyperthermia as stress agent for this experiment. Findings We here describe the expression of eight candidate reference genes from Atlantic cod (Gadus morhua l.) and their stability during thermal stress (temperature elevation of one degree C/day for 5 days). The genes investigated were: Eukaryotic elongation factor 1 alpha, ef1a; 18s ribosomal RNA; 18s, Ubiquitin conjugate protein; ubiq, cytoskeletal beta-actin; actb, major histcompatibility complex I; MHC-I light chain, beta-2 -microglobulin; b2m, cytoskeletal alpha-tubulin; tba1c, acidic ribosomal phosphoprotein; rplp1, glucose-6-phosphate dehydrogenase; g6pd. Their expression were analyzed in 6 tissues (liver, head kidney, intestine, spleen, heart and gills) from cods exposed to elevated temperature and compared to a control group. Although there were variations between tissues with respect to reference gene stability, four transcripts were more consistent than the others: ubiq, ef1a, 18s and rplp1. We therefore used these to analyze the expression of stress related genes (heat shock proteins) induced during hyperthermia. We found that both transcripts were significantly upregulated in several tissues in fish exposed to increased temperature. Conclusion This is the first study comparing reference genes for RT-qPCR analyses of expression during hyperthermia in Atlantic cod. ef1a, 18s, rplp1 and ubiq transcripts were found to be well suited as reference genes during these experimental conditions. Article in Journal/Newspaper atlantic cod Gadus morhua Universitet i Oslo: Digitale utgivelser ved UiO (DUO) BMC Research Notes 4 1 |
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Open Polar |
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Universitet i Oslo: Digitale utgivelser ved UiO (DUO) |
op_collection_id |
ftoslouniv |
language |
English |
description |
Background One important physiological response to environmental stress in animals is change in gene expression. To obtain reliable data from gene expression studies using RT-qPCR it is important to evaluate a set of possible reference genes as normalizers for expression. The expression of these candidate genes should be analyzed in the relevant tissues during normal and stressed situations. To find suitable reference genes it was crucial that the genes were stably expressed also during a situation of physiological stress. For poikilotermic animals like cod, changes in temperature are normal, but if the changes are faster than physiological compensation, the animals respond with typical stress responses. It has previously been shown that Atlantic cod show stress responses when elevation of water temperature is faster than 1 degree/day, for this reason we chose hyperthermia as stress agent for this experiment. Findings We here describe the expression of eight candidate reference genes from Atlantic cod (Gadus morhua l.) and their stability during thermal stress (temperature elevation of one degree C/day for 5 days). The genes investigated were: Eukaryotic elongation factor 1 alpha, ef1a; 18s ribosomal RNA; 18s, Ubiquitin conjugate protein; ubiq, cytoskeletal beta-actin; actb, major histcompatibility complex I; MHC-I light chain, beta-2 -microglobulin; b2m, cytoskeletal alpha-tubulin; tba1c, acidic ribosomal phosphoprotein; rplp1, glucose-6-phosphate dehydrogenase; g6pd. Their expression were analyzed in 6 tissues (liver, head kidney, intestine, spleen, heart and gills) from cods exposed to elevated temperature and compared to a control group. Although there were variations between tissues with respect to reference gene stability, four transcripts were more consistent than the others: ubiq, ef1a, 18s and rplp1. We therefore used these to analyze the expression of stress related genes (heat shock proteins) induced during hyperthermia. We found that both transcripts were significantly upregulated in several tissues in fish exposed to increased temperature. Conclusion This is the first study comparing reference genes for RT-qPCR analyses of expression during hyperthermia in Atlantic cod. ef1a, 18s, rplp1 and ubiq transcripts were found to be well suited as reference genes during these experimental conditions. |
format |
Article in Journal/Newspaper |
author |
Aursnes, Ingrid A Rishovd, Anne L Karlsen, Hans E Gjøen, Tor |
spellingShingle |
Aursnes, Ingrid A Rishovd, Anne L Karlsen, Hans E Gjøen, Tor Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress |
author_facet |
Aursnes, Ingrid A Rishovd, Anne L Karlsen, Hans E Gjøen, Tor |
author_sort |
Aursnes, Ingrid A |
title |
Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress |
title_short |
Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress |
title_full |
Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress |
title_fullStr |
Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress |
title_full_unstemmed |
Validation of reference genes for quantitative RT-qPCR studies of gene expression in Atlantic cod (Gadus morhua l.) during temperature stress |
title_sort |
validation of reference genes for quantitative rt-qpcr studies of gene expression in atlantic cod (gadus morhua l.) during temperature stress |
publishDate |
2011 |
url |
http://hdl.handle.net/10852/46293 http://urn.nb.no/URN:NBN:no-50541 https://doi.org/10.1186/1756-0500-4-104 |
genre |
atlantic cod Gadus morhua |
genre_facet |
atlantic cod Gadus morhua |
op_relation |
http://urn.nb.no/URN:NBN:no-50541 BMC Research Notes. 2011 Apr 05;4(1):104 http://hdl.handle.net/10852/46293 http://dx.doi.org/10.1186/1756-0500-4-104 URN:NBN:no-50541 Fulltext https://www.duo.uio.no/bitstream/handle/10852/46293/1/13104_2010_Article_856.pdf |
op_rights |
Gjøen et al; licensee BioMed Central Ltd. Attribution 2.0 Generic http://creativecommons.org/licenses/by/2.0/ |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1186/1756-0500-4-104 |
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BMC Research Notes |
container_volume |
4 |
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1 |
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1766357524858011648 |