Kirschneck, Christian and Proff, Peter and Fanghaenel, Jochen and Wolf, Michael and Roldan, J. Camilo and Roemer, Piero (2016) Reference genes for valid gene expression studies on rat dental, periodontal and alveolar bone tissue by means of RT-qPCR with a focus on orthodontic tooth movement and periodontitis. ANNALS OF ANATOMY-ANATOMISCHER ANZEIGER, 204. pp. 93-105. ISSN 0940-9602, 1618-0402
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Objectives: To obtain valid results in relative gene/mRNA-expression analyses by RT-qPCR, a careful selection of stable reference genes is required for normalization. Currently there is little information on reference gene stability in dental, periodontal and alveolar bone tissues of the rat, especially regarding orthodontic tooth movement and periodontitis. We therefore aimed to identify the best selection and number of reference genes under these experimental as well as physiological conditions. Materials and methods: In 7 male Fischer344-rats the upper left first and second molars were moved orthodontically for 2 weeks and in 7 more animals additionally subjected to an experimental periodontitis, whereas 7 animals were left untreated. Tissue samples of defined size containing both molars (without crowns) as well as the adjacent periodontal and alveolar bone tissue were retrieved and RNA extracted for RT-qPCR analyses. Nine candidate reference genes were evaluated and ranked according to their expression stability by 4 different algorithms (geNorm, NormFinder, BestKeeper, comparative Delta C-q). Results: PPIB/YWHAZ were the most stabile reference genes for the combined dental, periodontal and alveolar bone tissue of the rat overall, in untreated animals and rats with additional periodontitis, whereas PPIB/B2M performed best in orthodontically treated rats with YWHAZ ranking third. Gene-stability ranking differed considerably between investigated groups. A combination of two reference genes was found to be sufficient for normalization in all cases. Conclusions: The substantial differences in expression stability emphasize the need for valid reference genes, when aiming for meaningful results in relative gene expression analyses. Our results should enable researchers to optimize gene expression analysis in future studies by choosing the most suitable reference genes for normalization. (C) 2015 Published by Elsevier GmbH.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | REAL-TIME PCR; HOUSEKEEPING GENES; QUANTITATIVE PCR; IN-VIVO; MODEL; SELECTION; NORMALIZATION; IDENTIFICATION; CELLS; INJURY; RT-qPCR; Gene expression; Gene expression regulation; Rats, Inbred F344; Orthodontics; Periodontitis; Dental research |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Mund-, Kiefer- und Gesichtschirurgie Medicine > Lehrstuhl für Zahnärztliche Prothetik |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 15 Mar 2019 11:23 |
| Last Modified: | 15 Mar 2019 11:23 |
| URI: | https://pred.uni-regensburg.de/id/eprint/2741 |
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