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 (avtor), Drevenšek, Gorazd (avtor), Drevenšek, Martina (avtor), Marc, Janja (avtor), Prodan Žitnik, Irena (avtor)

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.

• Jezik: Angleški jezik
• Ključne besede: reference genes, alveolar bone, RT-qPCR, rat, olanzapine
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: FFA - Fakulteta za farmacijo ; MF - Medicinska fakulteta
• Različica publikacije: Objavljena publikacija
• Leto izida: 2024
• Št. strani: Str. 511-524
• Številčenje: Vol. 74, no. 3
• PID: 20.500.12556/RUL-163546
• UDK: 602.64:616
• ISSN pri članku: 1846-9558
• DOI: 10.2478/acph-2024-0029
• COBISS.SI-ID: 206027523
• Datum objave v RUL: 08.10.2024

Gradivo je del revije

• Naslov: Acta pharmaceutica
• Skrajšan naslov: Acta pharm.
• Založnik: Croatian Pharmaceutical Society
• ISSN: 1846-9558
• COBISS.SI-ID: 3817585

Licence

• Licenca: CC BY-ND 4.0, Creative Commons Priznanje avtorstva-Brez predelav 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by-nd/4.0/deed.sl
• Opis: Licenca Creative Commons Brez predelav dovoljuje uporabnikom ponovno distribucijo dela, vendar ne v spremenjeni obliki. Zahtevana je navedba avtorstva.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: referenčni geni, alveolarna kost, RT-qPCR, podgane, olanzapin, genska terapija, gensko izražanje

Projekti

• Financer: ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
• Številka projekta: P3-0293
• Naslov: Parodontalna medicina

• Financer: Drugi - Drug financer ali več financerjev
• Številka projekta: AD FUTURA
• Naslov: AD FUTURA. Science and Education Foundation of the Republic of Slovenia,