PPIA, HRPT1, and PGK1 genes as the appropriate combination for RT-qPCR normalization in alveolar and femoral bone remodeling in olanzapine-treated rats

Disha Ibrahimi, Saranda (author), Drevenšek, Gorazd (author), Drevenšek, Martina (author), Marc, Janja (author), Prodan Žitnik, Irena (author)

Reliable gene expression analysis in bone remodeling studies requires an appropriate selection of internal controls, i.e. stable reference genes for the normalization of quantitative real-time PCR (RT-qPCR), the most common method used for quantifying gene expression measurements. Even the most widely used reference genes can have variable expression under different experimental conditions, or in different tissue types or treatment regimes, so selecting appropriate controls is a key step in ensuring reliable results. The aim of this research was to identify the most stable reference gene(s) for the study of olanzapine modulated bone remodeling in rats. RNA was isolated from the maxillary alveolar and femoral bones of olanzapine or placebo-treated Wistar rats and transcribed to cDNA. The expression of 12 candidate reference genes was assessed by RT‐qPCR. Their expressions were analysed using GeNorm, NormFinder, BestKeeper and delta Ct algorithms, and by the comprehensive ranking method. PPIA, HRPT1 and PGK1 were the most stably expressed reference genes and the combination of the three genes was optimal for normalization. This study is the first to identify the optimal reference genes for research in olanzapine-exposed rats, which serve as a pivotal benchmark for enhancing the accuracy and reliability of future RT-qPCR expression in bone studies.

• Language: English
• Keywords: reference genes, alveolar bone, RT-qPCR, rat, olanzapine
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy; MF - Faculty of Medicine
• Publication version: Version of Record
• Year: 2024
• Number of pages: Str. 511-524
• Numbering: Vol. 74, no. 3
• PID: 20.500.12556/RUL-163546
• UDC: 602.64:616
• ISSN on article: 1846-9558
• DOI: 10.2478/acph-2024-0029
• COBISS.SI-ID: 206027523
• Publication date in RUL: 08.10.2024

Record is a part of a journal

• Title: Acta pharmaceutica
• Shortened title: Acta pharm.
• Publisher: Croatian Pharmaceutical Society
• ISSN: 1846-9558
• COBISS.SI-ID: 3817585

Licences

• License: CC BY-ND 4.0, Creative Commons Attribution-NoDerivatives 4.0 International
• Link: http://creativecommons.org/licenses/by-nd/4.0/
• Description: Under the NoDerivatives Creative Commons license one can take a work released under this license and re-distribute it, but it cannot be shared with others in adapted form, and credit must be provided to the author.

Secondary language

• Language: Slovenian
• Keywords: referenčni geni, alveolarna kost, RT-qPCR, podgane, olanzapin, genska terapija, gensko izražanje

Projects

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: P3-0293
• Name: Parodontalna medicina

• Funder: Other - Other funder or multiple funders
• Project number: AD FUTURA
• Name: AD FUTURA. Science and Education Foundation of the Republic of Slovenia,