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http://hdl.handle.net/1942/18153
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DC Field | Value | Language |
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dc.contributor.author | RAVINDRAN NAIR, Ambily | - |
dc.contributor.author | SMEETS, Karen | - |
dc.contributor.author | KEUNEN, Els | - |
dc.contributor.author | Lee, Wing-Kee | - |
dc.contributor.author | Thévenod, Frank | - |
dc.contributor.author | VAN KERKHOVE, Emmy | - |
dc.contributor.author | CUYPERS, Ann | - |
dc.date.accessioned | 2015-01-21T14:49:18Z | - |
dc.date.available | 2015-01-21T14:49:18Z | - |
dc.date.issued | 2015 | - |
dc.identifier.citation | JOURNAL OF APPLIED TOXICOLOGY, 35 (5), p. 478-484 | - |
dc.identifier.issn | 0260-437X | - |
dc.identifier.uri | http://hdl.handle.net/1942/18153 | - |
dc.description.abstract | Cadmium (Cd) is a toxic metal with a long half-life in biological systems. This half-life is partly as a result of metallothioneins (MTs), metal-binding proteins with a high affinity for Cd. The high retention properties of the kidneys reside in proximal tubular cells that possess transport mechanisms for Cd-MT uptake, ultimately leading to more Cd accumulation. Researchers have studied MT–metal interactions using various techniques including quantitative real-time PCR (qPCR), an efficient tool for quantifying gene expression. Often a poor choice of reference genes, which is represented by their instability and condition dependency, leads to inefficient normalization of gene expression data and misinterpretations. This study demonstrates the importance of an efficient normalization strategy in toxicological research. A selection of stable reference genes was proposed in order to acquire reliable and reproducible gene quantification under metal stress using MT expression as an example. Moreover, in vitro and in vivo setups were compared to identify the influence of toxicological compounds in function of the experimental design. This study shows that glyceraldehyde-3-phosphate dehydrogenase (Gapdh), tyrosine monooxygenase/tryptophan5-monooxygenase activation-protein, zeta polypeptide (Ywhaz) and betaactin (Actb) are the most stable reference genes in a kidney proximal tubular cell line exposed to moderate and high Cd concentrations, applied as CdCl2. A slightly different sequence in reference gene stability was found in renal cells isolated from rats in vivo exposed to Cd. It was further shown that three reference genes are required for efficient normalization in this experimental setup. This study demonstrates the importance of an efficient normalization strategy in toxicological research. | - |
dc.description.sponsorship | The authors would like to thank Rosette Beenaerts, Biomedical Research Institute, Hasselt University for her technical assistance and Dr Michael D. Garrick at the University of Buffalo for the Fischer 344 rats. This work was supported by Hasselt University [BOF (Bijzonder onderzoeksfonds) projects; BOF08G01] through a PhD grant for Ambily Ravindran Nair. Additional funding came from tUL-impulsfinanciering (project toxicology), and Methusalem project (08M03VGRJ). Frank Thevenod's research was supported by the Deutsche Forschungsgemeinschaft (DFG TH345/11-1) and by the Centre for Biomedical Training and Research (ZBAF) of the University of Witten/Herdecke. | - |
dc.language.iso | en | - |
dc.rights | Copyright © 2014 John Wiley & Sons, Ltd. | - |
dc.subject.other | cadmium; metallothionein; qPCR; reference genes | - |
dc.title | Renal cells exposed to cadmium in vitro and in vivo: normalizing gene expression data | - |
dc.type | Journal Contribution | - |
dc.identifier.epage | 484 | - |
dc.identifier.issue | 5 | - |
dc.identifier.spage | 478 | - |
dc.identifier.volume | 35 | - |
local.format.pages | 7 | - |
local.bibliographicCitation.jcat | A1 | - |
dc.description.notes | Smeets, K (reprint author), Hasselt Univ, Ctr Environm Sci, Agoralaan Bldg D, B-3590 Diepenbeek, Belgium. karen.smeets@uhasselt.be | - |
local.type.refereed | Refereed | - |
local.type.specified | Article | - |
dc.identifier.doi | 10.1002/jat.3047 | - |
dc.identifier.isi | 000351684300004 | - |
item.contributor | RAVINDRAN NAIR, Ambily | - |
item.contributor | SMEETS, Karen | - |
item.contributor | KEUNEN, Els | - |
item.contributor | Lee, Wing-Kee | - |
item.contributor | Thévenod, Frank | - |
item.contributor | VAN KERKHOVE, Emmy | - |
item.contributor | CUYPERS, Ann | - |
item.fullcitation | RAVINDRAN NAIR, Ambily; SMEETS, Karen; KEUNEN, Els; Lee, Wing-Kee; Thévenod, Frank; VAN KERKHOVE, Emmy & CUYPERS, Ann (2015) Renal cells exposed to cadmium in vitro and in vivo: normalizing gene expression data. In: JOURNAL OF APPLIED TOXICOLOGY, 35 (5), p. 478-484. | - |
item.accessRights | Restricted Access | - |
item.fulltext | With Fulltext | - |
item.validation | ecoom 2016 | - |
crisitem.journal.issn | 0260-437X | - |
crisitem.journal.eissn | 1099-1263 | - |
Appears in Collections: | Research publications |
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jat3047.pdf Restricted Access | Published version | 580.88 kB | Adobe PDF | View/Open Request a copy |
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